CN101370818A - RNAi inhibition of influenza virus replication - Google Patents

Abstract

The invention relates to compositions and methods for modulating the expression of influenza viral genes, and more particularly to the downregulation of influenza viral genes by chemically modified oligonucleotides.

Description

RNAi suppresses duplicating of influenza virus

Cross reference to related application

The right of priority of United States serial 60/748,317 that the application requires to submit on November 1st, 2005 U. S. application sequence number was submitted at December 7 in 60/732,243,2005 and the United States serial of submitting on May 9th, 2,006 60/799,000.The full content of each provisional application is incorporated this paper into by reference.

Technical field

The present invention relates to the treatment of influenza virus and the composition and the method field of adjusting virus replication, specifically, relate to disturbing and reduce the gene of influenza virus by oligonucleotide by RNA, described oligonucleotide is via suction/intranasal administration, be locally applied to lung and nasal meatus, or systemic administration, for example pass through via intravenous injection.

Background of invention

RNA disturbs or RNAi is a term, at first by Fire and its co-worker (Fire etc., Nature 391: 806-811,1998) propose to be used for to describe when with double-stranded RNA (dsRNA) but this observe phenomena of expression of its suppressor gene during the introducing worm.Comprise in the vertebrate life entity that many short dsRNA instructs the post-transcriptional silencing of gene specific, and provide new instrument for studying gene function.Recently this technology there is a lot of comments, consults, Novina for example, C.D., and Sharp, P., Nature 2004,430:161 and Sandy, P., etc., Biotechniques 2005,39:215, and it is by with reference to incorporating this paper into.

Influenza is that a kind of propagation is infected the most widely in the world wide.It can be fatal: at influenza A virus epidemic period in 1918, estimate at 2,000 ten thousand to 4,000 ten thousand people's death.In the U.S., annual about 20,000-40,000 people die from influenza a virus infection or its complication.When pestilence, the hospital care number that influenza is relevant just can reach more than 300,000 in single winter.

Several characteristic helps the success of influenza virus popular.At first, it propagates (droplet infection) easily by air between the crowd.Secondly, (antigenic drift) takes place in subtle change in the influenza antigen often, so that virus is easy to hide the inductive protective immunity by former different virus variant infection.The 3rd, by between different strains, genetic material being reset or being mixed, produce new influenza virus strain (antigenic shift) at an easy rate.If influenza A virus, this type of mixing can take place between hypotype that infects different plant species or strain.Pandemic disease in 1918 be it is said owing to causing derived from the viral hybridization strain that makes up between pig and influenza virus A hominis.At present, people generally worry to mix by the philtrum that contacts people and avian influenza virus at the same time, and latent infection people's new type influenza strain occurs, particularly from the influenza variant of fowl, more especially from strain H5N1.Be familiar with most of Asia society and had the expert of intimate contact to believe to poultry (agricultural birds) and their raiser, whether this mixing strain occurs not is a problem, and only is the problem that when takes place.The more serious pandemic disease of result than 1918 may be ensued apace.

Although strengthened effort, people still do not have effective therapy to influenza infection, and the partly cause that the value of existing vaccine is restricted is antigenic shift mentioned above and drift.For these reasons, influenza A virus has been carried out the whole world for many years and monitored, and National Institute of Health (National Institutes of Health) classifies it as one of pathogenic agent of biophylaxis override consideration.Although the present vaccine based on inactivation of viruses can be in about 70-80% age preventing disease in the healthy individual below 65 years old, this percentage ratio can reduce greatly in the individuality of the elderly or non-responsiveness.In addition, relevant with vaccine administration expense and potential side effect make that this method is undesirable.Although at present in being used for the treatment of of having got the Green Light of the U.S. and/or the antiviral of flu-prevention is helpful, owing to worry may occurring of side effect, compliance and resistance strain, their purposes is restricted.

U.S. Patent application 20040242518 and corresponding WO 04/028471, the both submitted on September 29th, 2003, had recommended the RNAi reagent that is used for the treatment of influenza of minority.Their effects in the mankind are not open.

Therefore, still need to research and develop the effective therapy that is used for the treatment of and prevents the virus infection of humans and animals, especially efficiently the target therapy of virus subtype on a large scale.A prerequisite is that activeconstituents is not degraded in physiological environment fast efficiently.

Summary of the invention

The present invention is based on external and intravital real example, can use this type of reagent and identify that from MP, NP, PB1, PB2 or the PA gene of influenza virus effective iRNA reagent suppresses influenza infection by intranasal administration iRNA reagent and by non-enteron aisle, described iRNA reagent can reduce the rna level of the many hypotypes of influenza virus.Find based on these, the invention provides concrete composition and method, it can be used for reducing for example virus titer of philtrum influenza virus mRNA level, influenza virus protein level and influenza virus of experimenter such as Mammals.

The present invention provides iRNA reagent specifically, it consists of, basic composition is or contains a kind of influenza virus gene, particularly at least 15 of a gene in influenza virus MP, NP, PB1, PB2 and the PA gene or more continuous nucleotide, more particularly, provide 15 or the reagent of more continuous nucleotides that contains from a kind of sequence shown in the table 1A-1H.Preferred each chain of iRNA reagent contains less than 30 Nucleotide, as 21-23 Nucleotide, as in showing shown in the 1A-1H.Double-stranded iRNA reagent can have flat terminal or more preferably have a overhang from one or two 3 ' terminal 1-4 Nucleotide of described reagent.

And iRNA reagent can only contain naturally occurring ribonucleotide subunit, perhaps can be synthetic so that contain one or more modifications that the sugar or the base of one or more ribonucleotide subunit are carried out in this reagent.Can further modify iRNA reagent, so that it is attached on the part that the stability, distribution or the cell that can selectivity increase this reagent absorb as cholesterol.IRNA reagent can also be isolating form, or the part of methods described herein pharmaceutical compositions for use, particularly as prepare for delivery to lung or nasal cavity or be used for that non-enteron aisle is used and the part of pharmaceutical compositions prepared.Described pharmaceutical composition can contain one or more iRNA reagent, and in some embodiments, contains two or more iRNA reagent, every kind of all different sequences or the directed different influenza virus gene of directed a kind of influenza virus gene.

On the one hand, the present invention relates to double chain oligonucleotide, it contains at least a non-natural base (nucleobase).In certain embodiments, non-natural base is difluoro toluene base (difluorotolyl), nitroindoline base (nitroindolyl), nitro-pyrrole base (nitropyrrolyl) or nitroimidazole base (nitroimidazolyl).In preferred embodiments, non-natural base is the difluoro toluene base.In certain embodiments, one that only contains in two oligonucleotide chains of double chain oligonucleotide contains the non-natural base.In certain embodiments, two oligonucleotide chains that contain double chain nucleotide all contain non-natural base alone.

The present invention also provides the method that reduces the viral RNA of influenza virus in the cell.This method comprise as described further below with a kind of iRNA agent administration of the present invention in experimenter's step.Present method utilize with degradation of cell optionally in the relevant cell mechanism of RNA interference of viral RNA, and comprise the step that cell is contacted with a kind of antiviral iRNA reagent of the present invention.Described method directly pair cell is carried out, and maybe can carry out on mammalian subject by use a kind of iRNA reagent/pharmaceutical composition of the present invention to the experimenter.The reduction of viral RNA causes the amount of the viral protein that produced to reduce in the cell, and in life entity, cause virus replication tire (replicating viral titer) reduce (as shown in the Examples).

Method and composition of the present invention, for example method and iRNA reagent composition, can any dosage shown in this article and/or formulation use, and use with any route of administration shown in this article.Particularly importantly, this paper intranasal administration of having shown iRNA reagent with and in respiratory tissue, suppress the ability of virus replication.

Being described in detail in hereinafter of one or more embodiments of the present invention proposes together in company with diagram and description.According to specification sheets, diagram and claims, other features of the present invention, target and advantage are conspicuous.For all purposes, the application introduces the full content of whole cited literature 2s, patent and patent application by reference.

Description of drawings

Figure 1A-1I: selected RNAi reagent suppresses the dose response curve of expression of target gene.To the Cos-7 cell, make the mRNA of this target gene of coding and Renilla luciferase express separately target gene recombinant clone in plasmid, described cell RNAi agent treated, and quantize with the Renilla luciferase.With cell concentration is the RNAi agent treated of 100nM, 25nM, 6.3nM, 1.6nM, 400pM, 100pM, 24pM, 6pM, 1.5pM and 380fM, and by parameterized fitting of a curve (parametrized curve fitting), utilize the XLfit program, determine IC ₅₀Value.

Detailed Description Of The Invention

Term " influenza virus " is used in reference to any strain of influenza virus in this article, and it can cause disease in animal or human experimenter, or it is the target candidate thing (interesting candidate) for analysis of experiments. Influenza virus is described in Fields, B., etc., Fields ＇ Virology, 4^thEd.2001, Lippincott Williams and Wilkins; Philadelphia is among the ISBN:0781718325. Particularly, this term comprises any strain of influenza A virus, and it can cause disease in animal or human experimenter, or it is the target candidate thing for analysis of experiments. A large amount of Flu-A separated strains has carried out completely or partially order-checking. It only is the part tabulation of Flu-A genomic fragment sufficient sequence shown in the table 6, it is stored in public database (influenza sequence library (The influenza Sequence Database, ISD)) in, consult Macken, C., Lu, H., Goodman, J., ﹠ Boykin, L., ＂ The value of a database in surveillance and vaccine selection. ＂ in Options for the Control of influenza IV.A.D.M.E.Osterhaus, N.Cox ﹠ A.W.Hampson (Eds.) 2001, Elsevier Science, Amsterdam, 103-106 page or leaf). This database also comprises B-mode and the sufficient sequence influenza C genomic fragment. This database can obtain on the internet, and comprises easily search engine, its can allow the user by genomic fragment, by virus infections kind and search for by the time of separating. The influenza sequence also can obtain at GenBank. Therefore, the influenza gene order can obtain expediently or be measured expediently by this area routine techniques personnel.

For convenience of explanation, for the single aggressiveness subunit of one or more RNA reagent, this paper uses term " nucleotides " or " ribonucleotide " sometimes. Should be appreciated that, the usage of term " ribonucleotide " or " nucleotides " herein, in the situation of the RNA that modifies or nucleotide substitution, also refer to the part that the nucleotides modified in one or more positions or substitute replace, as hereinafter further described.

As used herein, " RNA reagent " refers to the RNA of unmodified, RNA or the nucleosides substitute of modification, and every kind all describes at this paper, or the synthetic field of RNA is known. Although described RNAs and the nucleosides substitute of many modifications, preferred example comprises RNAs and the nucleosides substitute that has those modifications of stronger nuclease degradation resistance than the RNAs of unmodified. Preferred example comprise have 2 ' sugar-modified, strand jag (overhang) is modified (preferred 3 ' strand jag), or especially if in the situation of strand, comprise that 5 ' of one or more phosphates or one or more phosphate analogs modified.

As used herein, " iRNA reagent " (abbreviation of RNA interfering reagent) is RNA reagent, and it can reduce the expression of target gene such as influenza virus. Do not wish to be subject to theory, iRNA reagent can work by in many mechanism one or more, comprises the rear cutting of transcribing of said target mrna (being sometimes referred to as in the art RNAi), or transcribe front or translation before mechanism. IRNA reagent can be double-stranded iRNA reagent.

As used herein, " ds iRNA reagent " (abbreviation of double-stranded iRNA reagent) is the iRNA reagent that comprises more than preferred 2 chains, and wherein the hybridization of interchain can form the duplex structural region. In this article, " chain " refers to the nucleotides continuous sequence of (comprising nucleotides that non-natural exists or that modify). The two or more pieces chain can be independent molecule, and perhaps each forms the part of independent molecule, but perhaps their covalency for example interconnect, and by connector, for example the polyethylene glycol connector forms a molecule. At least one chain can comprise the zone fully complementary with target RNA. This class chain term is called " antisense strand ". The second chain of dsRNA reagent, it comprises the zone with the antisense strand complementation, and term is called " sense strand ". But ds iRNA also can form from the single rna molecule, and described single rna molecule has at least the part oneself complementary, forms the hair clip or the panhandle structure (panhandle structure) that for example comprise the duplex zone. The latter is referred to herein as short hairpin RNA s (short hairpin RNAs) or shRNAs. In the case, term " chain " refers in the RNA molecular domains, its with identical RNA molecule in another regional complementarity.

Although in mammalian cell, long ds iRNA reagent can be induced often harmful interferon response, and short ds iRNA reagent does not trigger interferon response, at least not to degree (Manche etc., Mol.Cell.Biol. that cell and/or host are harmful to12: 5238,1992; Lee etc., Virology199: 491,1994; Castelli etc., J.Exp.Med.186: 967,1997; Zheng etc., RNA10: 1934,2004; Heidel etc., ＂ Lack of interferon response in animals to naked siRNAs ＂ Nature Biotechn. advance online publication doi:10.1038/nbtl038, Nov.21,2004). IRNA reagent of the present invention comprises enough weak points so that can not trigger the molecule that harmful nonspecific interference element is replied in normal mammalian cell. Therefore, use the composition that comprises iRNA reagent (for example as described herein and preparation) to the experimenter and be used in the expression that reduces influenza virus gene in the experimenter influenza virus express cell, and the avoid interference element is replied. Enough short so that can not trigger the molecule that the harmful interference element replys and be referred to herein as siRNA reagent or siRNAs. As used herein, " siRNA reagent " or " siRNA " refer to iRNA reagent, ds iRNA reagent for example, it is enough lacked so that can not particularly induce harmful interferon response in people's cell from mammal, and for example it has less than 60 but preferred duplex zone less than 50,40 or 30 nucleotide pairs.

As described herein, the iRNA reagent of separation comprises ds iRNA reagent and siRNA reagent, can mediate influenza nucleic acids and express reduction, for example by RNA degraded mediation. For convenience, this RNA is also referred to as in this article and treats reticent RNA. This nucleic acid is also referred to as target gene. Preferably, treat that reticent RNA is the gene outcome of influenza virus gene, described gene outcome is the part of influenza virus strain that the people is caused a disease.

As used herein, phrase " mediate rna i " refers to that reagent makes the ability of target gene silence in sequence-specific mode." make the target gene silence " and mean such process, promptly compare with the similar cell that does not contact reagent, contain and/or express the cell of some target gene product, when not contacting, can contain and/or express at least and to lack 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90% described gene product when contacting with this reagent with reagent.Described target gene product can be for example messenger RNA(mRNA) (mRNA), protein or regulatory element.

As used herein, term " complementary " is used to represent complementary enough degree, so that combination stable and that for example take place between the influenza virus mRNA at compound of the present invention and target RNA molecule specifically.Specificity is in conjunction with the complementarity of the enough degree of needs, take place under needs specificity bonded condition with the non-specific binding of avoiding non-target sequence and oligomeric compounds, promptly, analyze in vivo or treat under the physiological condition under the disposition, or under the situation of analyzed in vitro, under the condition of carrying out this analysis.Non-target sequence is usually by having at least 2,3 or 4 Nucleotide to be different from target sequence.

As used herein, if iRNA reagent has reduced the generation of target RNA encoded protein in cell, iRNA reagent and target RNA said target mrna (as target influenza virus mRNA) " fully complementary " for example then.IRNA reagent also can with target RNA " accurately complementary ", for example, target RNA and the annealing of iRNA reagent are preferably to form the heterocomplex that is made of the Watson-Crick base pair fully in accurate complementary region." fully complementary " iRNA reagent can comprise and the accurate complementary interior region of target Influenza Virus RNA (for example interior region of 10 Nucleotide) at least.And in some embodiments, iRNA reagent can be distinguished the difference of mononucleotide specifically.In the case, if accurately complementation is found in the zone (for example in 7 Nucleotide) of mononucleotide difference, iRNA reagent mediate rna i only then.Preferred iRNA reagent based on or contain provide among the table 1A-1H adopted sequence and antisense sequences arranged, or form by it.

As used herein, " substantially the same " is when being used in reference to article one nucleotide sequence that compares with the second nucleotide sequence, it is outer identical with the second nucleotide sequence except having 1,2 or 3 Nucleotide to replace (for example, uridylic has replaced adenosine) at most to mean article one nucleotide sequence.As used herein, for being different from but by disappearance, adding or the replacement of Nucleotide derived from the table 1A-1H iRNA reagent in a kind of iRNA reagent, " be retained in basically and suppress the ability that influenza virus is expressed in human influenza virus's express cell of cultivation ", the meaning be meant inhibition activity that deutero-iRNA reagent has be at least table 1A-1H its derived from the inhibition active 20% of iRNA reagent.For example, reduce by 70% iRNA reagent deutero-iRNA reagent from the amount that can make the influenza virus mRNA that the people's cell of cultivating that is subjected to influenza infection, exists of table 1A-1H, can be separately the amount of the influenza virus mRNA that exists be reduced by 50% in the people's cell of cultivating that is subjected to influenza infection, thereby think to have kept basically and in the people's cell of cultivating that is subjected to influenza infection, suppress the ability that influenza virus is duplicated.Randomly, iRNA reagent of the present invention can reduce at least 50% with the amount of the influenza virus mRNA that exists in the people's cell of cultivating that is subjected to influenza infection.

As used herein, " experimenter " makes a comment or criticism and accepts Mammals life entity by the disease treatment of influenza infection mediation.The experimenter can be any Mammals, as ox, horse, mouse, rat, dog, pig, goat or primates.In preferred embodiments, the experimenter is the people.

The feature of influenza virus

Influenza virus is the coating minus-stranded rna virus of Orthomyxoviridae family (Orthomyxoviridae).They are divided into first type, B-mode and influenza C, and wherein influenza A is that tool is pathogenic and be considered to the unique type that can reset with the animal strain.First type, B-mode and influenza C can be distinguished by the difference in nucleoprotein and the stromatin.As further discussed, the influenza A hypotype defines by their hemagglutinin (HA) or the sudden change in Sialidase (NA) gene, and usually by distinguishing with corresponding protein bonded antibody.

The influenza A virus genome is by 10 genomic constitutions that are distributed in 8 RNA fragments.10 kinds of albumen of this genes encoding: envelope glycoprotein hemagglutinin (HA) and Sialidase (NA); Stromatin (being called M1 or MP herein); Nucleoprotein (NP); Three kinds of polysaccharases (PB1, PB2 and PA), it is the constituent of RNA RNA-dependent transcriptase, is also referred to as polysaccharase or polysaccharase complex body in this article; Ionophorous protein (M2), and Nonstructural Protein (NS1 and NS2).Relevant influenza A virus with and further describing of molecular pathogenesis consult Julkunen, I., etc., Cytokine and GrowthFactor Reviews, 12:171-180,2001.Also can consult Fields, B., etc., Fields ＇ Virology, 4.sup.th.ed., Philadelphia:Lippincott Williams and Wilkins; ISBN:0781718325,2001.Genomic composition of Influenza B virus and influenza A are closely similar, but the influenza virus C genome contains 7 kinds of RNA fragments and lacks the NA gene.

The classification of influenza A virus is a foundation with hemagglutinin (H1-H15) and Sialidase (N1-N9) gene.World health organization (World Health Organization, WHO) nomenclature according to the animal host of viral source (specifying, only the people), geographic origin, strain numbering, separate time and HA and NA antigenicity and describe and define each virus strain.For example, A/Puerto Rico/8/34 (H1N1) expression first type strain, isolate 8 and had HA and NA antigenicity hypotype in its 1934 on the person that betides Puerto Rico (Puerto Rico).Another example A/Chicken/Hong Kong/258/97 (H5N1), expression first type strain, isolate 258 betided on one's body the chicken (chicken) in Hong Kong, and have the antigenicity hypotype 5 of HA and the antigenicity hypotype 1 of NA in its 1997.The virus that contains HA type H1, H2 and H3 and NA type N1 and N2 has caused human prevailing disease.

As indicated above, in influenza A virus, heritable variation takes place by two kinds of main mechanism.Antigenic drift takes place via point mutation, because selection pressure from host immune response, it often occurs in the position with antigenicity meaning, antigenic shift (being also referred to as rearrangement), and the whole viral genome fragment that relates to a kind of hypotype is replaced by another kind.Many dissimilar animal species comprise the mankind, pig, bird, horse, aquatic mammal and other kinds, all may be subjected to influenza a virus infection.Some influenza A viruss are subject to specific species, and do not infect different species usually.Yet some influenza A viruss can infect several different animal species, mainly are bird (particularly Qian Yi aquatic bird), pig and people.This ability is considered to be in the reason that forms most antigenic shifts in the influenza A virus.For example, suppose that pig has infected the influenza A virus from the people, and infected another kind of influenza A virus simultaneously from duck (duck).When these two kinds of different viruses were bred in pig cell, the gene of the gene of people's strain and duck strain just can " mix ", thereby produced the new virus with unique RNA fragment combination.This process is called gene rearrangement (note: such gene rearrangement betides the exchange of the genetic information between the karyomit(e) when being different from reduction division).

The same with some bacterial species with other virus, influenza virus is a time multiplexed cell system.Influenza A virus duplicates in the epithelial cell of the upper respiratory tract.Yet, also can infect monocyte/macrophage and other white corpuscle.Many other have contain sialic cell surface glycoprotein cell type all easily external infected, because virus utilizes these molecules as acceptor.

The design of iRNA reagent and selection

As used herein, " expressing relevant disease with influenza virus " refers to any biology or pathologic state, and the influenza virus that its (1) to small part is existed mediates, and (2) influenza virus exists the reduction of level can influence their final result.Expressing relevant disease specific with influenza virus mentions hereinafter.

The present invention is based on design, synthesize and produce the iRNA reagent of the virogene of energy target influenza virus; also based on after using iRNA reagent incubation; make the virogene silence external in cultured cells, the proof, and the protective effect to virus infection that is obtained.

Based on sequence information and desired feature, can reasonably design iRNA reagent.For example, can be according to the relative melting temperature(Tm) design iRNA of candidate's duplex.Usually, 5 ' of duplex antisense strand end has lower melting temperature(Tm) than 3 ' end at antisense strand.

The invention provides the siRNA (s) that contains one or more influenza virus transcripts of target and/or the composition of shRNA (s).Because the description of above-mentioned influenza virus replication cycle has proved various types of viral RNA transcript (firsts and seconds vRNA, firsts and seconds virus mRNA and viral cRNA) all be present in the cell that has infected influenza virus, and in the life cycle of virus, play an important role.Any of these transcript all be siRNA by according to of the present invention directly and indirect mechanism mediate the suitable target of inhibition.The siRNAs of any virus mRNA transcript of target and shRNAs promptly reduce the transcript own level by transcript is degraded specifically in direct mode.In addition, as discussed below, siRNAs of some virus transcription thing of target (as MP, PA, PB1) and shRNAs can make the virus transcription thing level of its non-specific target reduce indirectly.In optional montage is under the contingent situation, and for the mRNA of coding MP and M2 and the mRNA of coding NS1 and NS2, the transcript of the transcript of montage or montage all can not serve as target transcript (target transcript).

The potential transcript, the target that it can serve as according to the therapy based on RNAi of the present invention for example comprises 1) any influenza virus gene group fragment; 2) transcript of any viral protein of coding comprises the transcript of proteins encoded PB1, PB2, PA, NP, NS1, NS2, MP, M2, HA or NA.Should be appreciated that, but the transcript of single siRNA or shRNA target vRNA, cRNA and/or mRNA form.Yet as Ge etc., WO 04/028471 is proposed, and perhaps virus mRNA is the unique or main target of RNAi.

For any concrete gene target of selecting, design is preferably adopted some guilding principle according to the present invention used siRNA or shRNA.Generally speaking, the desired target sequence is specific (comparing with the host) to virus, and, be important or necessary preferably to viral function.Although some virogene, the feature of those genes of particularly encode HA and NA are mutation rate height and can stand sudden change that some zone and/or sequence are still tended to guard.According to certain embodiments of the present invention, this type of sequence is particularly suitable target.As described further below, this type of conservative region can much all be that the public is obtainable by consulting document and/or convection current sensillary base because of sequence compares to determine.And, in many cases, be delivered to the reagent of cell of the present invention, can before becoming activity inhibitor, carry out a step or multistep treatment step (further discuss and consult hereinafter); In the case, it will be understood by those skilled in the art that preferably related reagent being designed to comprise its treatment step is the sequence of necessity.An aspect of of the present present invention is to recognize (being referred to as variant) such as a plurality of strains when infectious media, hypotypes when existing, and its genome sequence changes, and just often the siRNAs and the shRNAs in the different variant high conservative of target zone are selected and/or design in expectation.Particularly, by sequence that compares sufficient amount and the zone of selecting high conservative, be exactly possible with a plurality of variants of single siRNA target so, the duplex of described siRNA partly comprises described high conservative zone.Usually, should there be enough length in described zone, so that comprise the whole duplexs parts (for example 19 Nucleotide) of siRNA and randomly, one or more 3 ' overhangs are although (for example 15,16,17 or 18 Nucleotide) also can be used in the zone shorter than the total length of duplex.According to certain embodiments of the present invention, if a zone is identical between variant, then it is exactly high conservative between a plurality of variants.According to certain embodiments of the present invention, if zone (no matter length whether be included in siRNA the duplex part for example 15,16,17,18 or preferred 19 Nucleotide in) 1 Nucleotide (i.e. 0 or 1 Nucleotide) difference is arranged at most between variant, then it is a high conservative.According to certain embodiments of the present invention, if a zone has 2 Nucleotide differences (i.e. 0,1 or 2 Nucleotide) at most between variant, then this zone is a high conservative.According to certain embodiments of the present invention, if a zone has 3 Nucleotide differences (i.e. 0,1,2 or 3 Nucleotide) at most between variant, then this zone is a high conservative.According to certain embodiments of the present invention, siRNA comprises the duplex part in a zone of target, and described zone is a high conservative between at least 5 variants, at least 10 variants, at least 15 variants, at least 20 variants, at least 25 variants, at least 30 variants, at least 40 variants or at least 50 variants or more changeable body.

In order to determine whether a zone is high conservative, can make in the following method between multiple variant.Select a member in this group sequence as basic sequence (base sequence), that is, and the sequence that other sequences will compare with it.Usually, the length of basic sequence is the desired length of siRNA duplex part, for example 15,16,17,18 or preferred 19 Nucleotide.According to different embodiments of the present invention, basic sequence both can be a sequence that is just comparing in the group, also can be the deutero-consensus sequence, for example the consensus sequence by determining that the modal Nucleotide in each position derives in one group of sequence.

After selecting basic sequence, this each member's sequence of organizing in the multiple variant is all compared with basic sequence.Can be used for definite this basis sequence and this member high conservative whether on the concrete zone of target at the difference number of this sequence area between any member in basis sequence and the multiple variant of this group.As mentioned above, in various embodiments of the present invention, if the sequence difference number between two zones is 0; 0 or 1,0,1 or 2; Or 0,1,2 or 3, think that then this zone is a high conservative.In the position that difference occurs, can select the siRNA sequence, so that identical with a kind of sequence in this basis sequence or other sequences.Usually, select the Nucleotide that is present in the basic sequence.But in certain embodiments of the invention, find in more comparative sequences than the Nucleotide in the basic sequence, then can select the SiRNA identical with second sequence if particularly be present in the Nucleotide of specific position in second sequence that compares group.In addition, according to certain embodiments of the present invention,, then can use this total Nucleotide if be different from the Nucleotide of in basic sequence, finding at the total Nucleotide (the most ubiquitous Nucleotide) of difference nidus.Note, so just may make the sequence of acquisition be different from all sequences (as using consensus sequence) that is just comparing as basic sequence.

The inventor finds that most of sequence of using design variable hereinafter described to select can both suppress virus replication effectively after introducing siRNA or shRNA, and as mentioned below the test.

Based on these results shown in this article, the invention provides can be in the cell of cultivating that is subjected to influenza infection and experimenter such as Mammals for example among the people, reduces the iRNA reagent that influenza virus is duplicated.Table 1A-1H provides the exemplary iRNA reagent of target influenza virus.Table 1A, C, D and E have enumerated the phosphorothioate bond between 3 ' end and penult thymidine, do not contain the siRNAs of nucleotide modification.Table 1B and H have enumerated siRNAs, wherein all Nucleotide that contain pyrimidine base are the Nucleotide of 2 ＇-O-methyl-modification in sense strand, and the cytidine in the uridine in all 5 ＇-ua-3 ＇ sequence background and all 5 ＇-ca-3 ＇ sequence background is the Nucleotide of 2 ＇-O-methyl-modification in antisense strand, except having the AL-DP-2295 that duplex is determined, outside the iRNA of AL-DP-2301 and AL-DP-2302, wherein the uridine in all 5 ＇-ug-3 ＇ sequence background is the Nucleotide of 2 ＇-O-methyl-modification.These siRNAs of back are occurrence sequence primitive 5 ＇-ua-3 ＇ or 5 ＇-ca-3 ＇ not, and with these reagent in mice serum behind the incubation, to the analysis revealed of these degradation fragments, sequence motif 5 ＇-ug-3 ＇ is the main site that endonuclease is attacked.

Based on these results, this aspect provides the iRNA that comprises sense strand and antisense strand reagent especially, described sense strand has at least 15 successive Nucleotide of the sense strand sequence of reagent shown in the table 1A-1H, and described antisense strand has at least 15 successive Nucleotide of the antisense strand of reagent shown in the table 1A-1H.

Being shown in the iRNA reagent of table among the 1A-1H is that two chains of 19 Nucleotide are formed by length, and described two chains and target complement sequence or identical add 3 ＇-T TOverhang.The invention provides the reagent that contains at least 15 or at least 16,17 or 18 or 19 continuous nucleotides in these sequences.Yet,, and do not mean that iRNA be limited to these length although these length may be best potentially.It will be recognized by those skilled in the art that shorter or longer iRNA reagent is effective too, because in the certain-length scope, effect is the function of nucleotide sequence rather than the length of sequence.For example, Yang, etc., PNAS 99: 9942-9947 (2002) has proved the similar effect of the iRNA reagent of length between 21 and 30 base pairs.Other people have also proved and can both make gene by minimum iRNA reagent to about 15 base pair length effectively reticent (Byrom is etc., " Inducing RNAi with siRNA Cocktailsgenerated by RNase III " Tech Notes 10 (1), Ambion, Inc., Austin, TX).

Therefore, the present invention can and dream up a part of sequence of selecting between 15-19 Nucleotide from the sequence that table 1A-1H provides, and is used for obtaining to provide derived from table 1A-1H the iRNA reagent of a kind of sequence of sequence.Alternatively, one or several Nucleotide can be joined in the table sequence that 1A-1H provided and go, or join in the reagent that contains from 15 continuous nucleotides of reagent one of in these reagent, preferably, but nonessential, with the Nucleotide that adds can with target gene each sequence complementary mode of influenza virus gene for example.For example, with preceding 15 Nucleotide and 8 Nucleotide combinations can in influenza virus mRNA, finding this sequence 5 ' end of a reagent, thereby obtain that the reagent that 23 Nucleotide are arranged in justice and the antisense strand is arranged.All these iRNA reagent of deriving all is included in the bright iRNA reagent of we, suppresses the ability that influenza virus is duplicated as long as they can be retained in the people's cell that is subjected to influenza infection of cultivation basically.

The antisense strand length of iRNA reagent should equal or 14,15,16,17,18,19,25,29,40 or 50 Nucleotide at least.It should be equal to or less than 60,50,40 or 30 Nucleotide at degree.Preferred length range is 15-30,17-25, a 19-23 and 19-21 Nucleotide.

The sense strand length of iRNA reagent should equal or 14,15,16,17,18,19,25,29,40 or 50 Nucleotide at least.Its length should be equal to or less than 60,50,40 or 30 Nucleotide.Preferred length range is 15-30,17-25, a 19-23 and 19-21 Nucleotide.

The double-stranded partial-length of iRNA reagent should equal or 15,16,17,18,19,20,21,22,23,24,25,29,40 or 50 nucleotide pairs at least.Its length should be equal to or less than 60,50,40 or 30 nucleotide pairs.Preferred length range is 15-30,17-25, a 19-23 and 19-21 nucleotide pair.

Usually, iRNA reagent of the present invention comprises and the abundant complementary of corresponding influenza virus gene zone, and wants sufficiently long with regard to Nucleotide, so that iRNA reagent or its fragment can mediate the downward modulation of influenza virus gene.Perfect complementation between iRNA reagent and the target gene not necessarily, but consistence must be enough so that iRNA reagent or its cleaved products can guide sequence specificity silences, for example by RNAi cutting Influenza Virus RNA.

Therefore, iRNA reagent of the present invention comprises the reagent that contains sense strand and antisense strand, each described sense strand and antisense strand contain the sequence of at least 16,17 or 18 Nucleotide, described sequence is as hereinafter defined, substantially the same with a sequence among the table 1A-1H, all distinguish no more than 1,2 or 3 Nucleotide except each chain and replaced, and kept the ability that the inhibition influenza virus is duplicated in the people's cell of cultivating that is subjected to influenza infection basically respectively by other Nucleotide (for example uridylic substituted adenosines).Therefore, it is identical with table at least 15 Nucleotide of a sequence among the 1A-1H that these reagent have, but relatively target Influenza Virus RNA sequence or justice is arranged and antisense strand between introduced 1,2 or 3 base mispairing.Mispairing with target Influenza Virus RNA sequence, the particularly mispairing in antisense strand, the zone is the most patient endways, if exist, in then preferred 1 zone or a plurality of zone endways, for example in 6,5,4 or 3 Nucleotide of 5 ' and/or 3 ' end, most preferably in 6,5,4 or 3 Nucleotide of sense strand 5 ' end or antisense strand 3 ' end.Sense strand only needs with antisense strand fully complementary, to keep the complete double-stranded feature of molecule.

Preferably, selecting has justice and antisense strand, so that iRNA reagent comprises strand or pairing region not at one or two end of molecule.Therefore, iRNA reagent contains sense strand and antisense strand, comprises overhang after the preferred pairing, one or two 5 ' or 3 ' overhang for example, but 3 ' overhang of preferred 2-3 Nucleotide.Most of embodiments have 3 ' overhang.Preferred siRNA reagent is in the one or both ends of this iRNA reagent, and having length is the strand overhang of 1-4 or preferred 2 or 3 Nucleotide, preferred 3 ' overhang.Overhang can be since chain than due to another chain length, or due to two chains of equal length interlock.The unpaired nucleotide that forms overhang can be a ribonucleotide, or deoxyribonucleotide, preferred thymidine.Preferably 5 ' end is carried out phosphorylation, or also can be not by phosphorylation.

The preferred length in the zone of duplexization is 15 to 30 Nucleotide, and 18,19,20,21,22 and 23 Nucleotide most preferably are for example in the scope of siRNA reagent as discussed above.SiRNA reagent can be similar to the natural Dicer processed products from long dsRNAs on length and structure.In one embodiment, two of siRNA reagent chains have carried out connecting as covalently bound.Hairpin structure or other single-stranded structure of required double-stranded region are provided, preferred 3 ' overhang, also within the scope of the invention.

The assessment of candidate iRNA reagent

As if above pointed, the invention provides the system of the siRNAs of the inhibitor that is used for determining to be used as influenza infection and/or duplicates.As mentioned above, because in cell, shRNAs is handled, can produce the siRNAs duplex part that has identical sequence with shRNAs backbone structure (stem structure), therefore, described system can be used for definite shRNAs that is used as the inhibitor of influenza infection equally.For purposes of illustration, this part refers to siRNAs, but described system also comprises corresponding shRNAs.Specifically, the present invention has proved the successful preparation of target virogene with the siRNAs that prevents or suppress virus infection and/or duplicate.Technology described herein and reagent can be used to design the potential novel siRNA s of other genes of target or gene region expediently, and are easy to detect their inhibition influenza infections as indicated above and/or the activity of duplicating.Expect that influenza virus will continue to suddenly change and reset, therefore expectation continues research and development and the siRNAs that tests new different targets.

In various embodiments of the present invention, before with influenza virus gene group or its part transfection, transfection is simultaneously or after the transfection (for example in the several minutes, in several hours or in maximum several days), or before with influenza infection, infect simultaneously or after infecting, by candidate siRNA (s) being introduced cell (as by exogenous using (exogeneous adminstration) or by instructing siRNA external source synthetic carrier or construct to introduce cell) or introducing experimental animal, test potential influenza virus inhibitor.Alternatively, the influenza virus inhibitor can be tested by candidate siRNA (s) being introduced cell or experimental animal, but described cell or experimental animal grown place (productively) are subjected to influenza infection (being that cell can produce progeny virus).Subsequently, assessment candidate siRNA (s) reduces one or more aspects feature of target transcript level and/or inhibition or compacting (suppress) virus life cycle such as virus replication, pathogenic and/or infective ability.For example, utilize method well known in the art, can assess the generation of virion and/or the generation of viral protein directly or indirectly.

Can compare sending the cell of siRNA composition of the present invention or experimental animal (test cell/animal) and the similar or comparable cell of not accepting the present composition or experimental animal (control cells/animal is not for example accepted siRNA or accepted cell/animal of contrast siRNA such as non-virus transcription thing of target such as GFP).Test cell/animal can compare the susceptibility that infects with control cells/animal the susceptibility of influenza infection.Can be in the test cell/animal relevant with control cells/animal, the relatively generation of viral protein and/or progeny virus.Can compare viral infection equally, duplicate, other indexs (indicia) such as pathogenic.The extracorporeal antivirus effect examination criteria can utilize the cytopathic effect that suppresses virus plaque, virus, and (cytopathic effect is CPF) with viral hemagglutinin or other albumen, inhibition viral yield etc.Can be visually and absorb by dyestuff and to measure CPE.Consult for example Sidwell, R.W. and Smee, D.F, " In vitro and in vivo assay systems for study of influenza virusinhibitors ＂ Antiviral Res 2000,48:1.Usually, test cell/animal and control cells/animal are from same species, and for cell, can be similar or identical cell types.For example, the cell that can relatively be from same cell.When test cell was primary cell, usually, control cells also should be a primary cell.Usually use identical influenza virus strain to come test cell/animal and control cells/animal are compared.

For example, can measure candidate siRNA in the following manner easily and suppress the ability that influenza virus produces: (i) candidate siRNA is delivered to cell (before the contact influenza virus, simultaneously or afterwards); (ii) utilize the hemagglutinin analysis to assess the generation of viral hemagglutinin; And the amount of the hemagglutinin that produced when not having siRNA of the amount of the hemagglutinin that is produced (iii) will have siRNA the time compares.(this test does not need to comprise the contrast that lacks siRNA, but can utilize in the past about the information in the amount that does not have the hemagglutinin that is produced when suppressing).The amount of hemagglutinin significantly reduction shows that virus produces reduction.This experiment can be used for testing the siRNA of target any virus transcript, and is not limited to the siRNAs of the transcript of target coding viral hemagglutinin.

The ability that candidate siRNA reduces target transcript level also can be assessed by for example utilizing measurement target transcript amounts such as Northern blotting, RNase protection analysis (nuclease protection assays), reverse transcription (RT)-PCR, real-time RT-PCR, microarray analysis.The ability that candidate siRNA suppresses to be produced by target transcript encoded polypeptides (transcribe or post-transcriptional level on), can utilize multiple method based on antibody to measure, described method includes but not limited to Western blotting, immunoassay, ELISA, flow cytometry, arrays of immobilized protein etc.Usually, can use any method of the amount of measuring target transcript or the coded polypeptide of target transcript.

Usually, the level that some preferred influenza virus inhibitor may exist when not having inhibitor relatively (for example in the same cell that lacks this inhibitor), the level of target transcript is reduced at least about 2 times, preferably at least about 4 times, more preferably at least about 8 times, at least about 16 times, at least about 64 times or even higher degree.Usually, some preferred influenza virus inhibitor can suppress virus replication, thereby make the level of duplicating low at least about 2 times in than the control cells that does not contain this inhibitor in containing the cell of this inhibitor, preferably at least about 4 times, more preferably at least about 8 times, at least about 16 times, at least about 64 times, at least about 100 times, at least about 200 times or even higher degree.

Some preferred influenza virus inhibitor can suppress duplicating of virus, thereby after using siRNA and cells infected, can prevent detectable virus titer development at least 24 hours, at least 36 hours, at least 48 hours or at least 60 hours.After using siRNA, some preferred influenza virus inhibitor can prevent (promptly being reduced to undetectable level) or reduce virus replication significantly at least 24 hours, at least 36 hours, at least 48 hours or at least 60 hours.Various embodiments according to this aspect, virus replication significantly reduce be reduced to the level that may occur when not having siRNA less than about 90%, be reduced to the level that may occur when not having siRNA less than about 75%, be reduced to the level that may occur when not having siRNA less than about 50%, be reduced to the level that may occur when not having siRNA less than about 25%, or be reduced to the level that may occur when not having siRNA less than about 10%.The reduction of virus replication can be measured by utilizing any suitable method, and described method includes but not limited to measure HA and tires.

IRNA stable reagent property testing, modification and test again

As when iRNA reagent is introduced in the subject, can carry out stability assessment to candidate iRNA reagent, for example, it is to the susceptibility of endonuclease or exonuclease cutting.Can using method determine and easily modified the particularly site of cutting, as the site of the component cutting found in the subject.Described method comprises separation and identifies at candidate iRNA reagent behind external separating bio medium such as serum, blood plasma, saliva, cerebrospinal fluid or cell or tissue homogenate incubation, or make contact in experimenter and the candidate iRNA reagent body after, the formed fragment of enriching most of candidate iRNA reagent degraded is determined the site that is easy to cut in view of the above.Described method for example, does not have any restriction, sees among the international application no PCT/US2005/018931 that submits to 27 days total Mays in 2005.

After the site that easily is cut is determined, can design and/or synthetic further iRNA reagent, wherein, make the potential cleavage site can resist cutting, for example modify, for example 2 ＇-O-methyl by introducing 2 ' at cleavage site.Can test the stability of this further iRNA reagent again, and this process can repeat, till finding that iRNA reagent shows desirable stability.

The body build-in test

Can be in animal model (for example Mammals, for example in mouse or rat), to functional test the in the body of determining to suppress the iRNA reagent that influenza virus gene expresses.For example, can be with the iRNA agent administration in animal, and iRNA reagent carried out duplicating or be reduced to small part by the biology of influenza virus mediation or the ability assessment of pathological process about its bio distribution, stability and its suppress influenza virus.

IRNA reagent can directly be applied to target tissue, inject as passing through, or iRNA reagent can same mode be applied to animal model with being applied to physiognomy.Preferably, iRNA reagent is delivered to experimenter's respiratory tract (airway), in nose.

Also can assess interior distribution of cell of iRNA.Described assessment comprises determines whether iRNA reagent is absorbed into cell.Described assessment also comprises the stability (as the transformation period) of determining iRNA.By use with traceable marker (as fluorescent mark such as fluorescein; Radio-labeling, as ³⁵S, ³²P, ³³P or ³H; Gold grain (gold particles); Or immunohistochemical antigen particles) the iRNA reagent of Lian Jieing can promote the assessment of iRNA reagent in the body.

Can assess the ability that iRNA reagent downward modulation influenza virus is duplicated.Can measure the level of influenza virus gene expression in vivo,, or be undertaken by isolation of RNA the tissue before or after contact iRNA reagent for example by in situ hybridization.For needing to sacrifice the situation of animal for obtaining tissue, can be with untreated control animal as contrasting.Influenza Virus RNA can detect by any desired method, and described method includes but not limited to PR-PCR, Northern trace, branched DNA analysis or rnase protection analysis.Alternatively, or additionally, influenza virus gene is expressed and can be formed detection and monitor by the tissue extract with the iRNA agent treated being carried out Western engram analysis or plaque.

Utilize any model in the various animal models of having set up, can test potential influenza virus inhibitor.Before with influenza infection, simultaneously or afterwards, composition is applied to animal, and described composition contains candidate siRNA (s), can instruct described siRNA synthetic construct or carrier at host cell or in the cell that contains candidate siRNA through transforming or operating.Composition prophylaxis of viral infections and/or delay or the related indication appearance of flu-prevention and/or the ability that alleviates the seriousness of the animal that is subjected to influenza infection of not accepting potential influenza virus inhibitor are assessed.Described model includes but not limited to mouse (murine), chicken, ferret (ferret) and is used for the non-human primate model of influenza infection that they all are known, and the effect that can be used for testing potential influenza therapy and vaccine in this area.For example consult the above Sidwell of reference, R.W. and Smee, D.F.The strain (as WSN or PR8 strain, it is adapted at duplicating in the mouse) that described model can use naturally occurring influenza virus strain and/or modify or be adapted at surviving in the specific host.Above animal model also can be used to be defined as realizing certain desired effects and required concentration (as EC50).

The iRNA chemical property

Described isolating iRNA reagent at this, for example mediate rna i is to suppress the dsRNA reagent that influenza virus gene is expressed.

RNA reagent discussed herein comprises the RNA of other aspect unmodifieds and modified to improve the RNA of its effect, also comprises the polymer of nucleosides surrogate.The composition that the RNA of unmodified refers to nucleic acid promptly sugar, base and phosphoric acid part with naturally occurring, preferably with human body in the identical or substantially the same molecule of naturally occurring composition.In the prior art rare or uncommon but naturally occurring RNA is called the RNA of modification, this content is for example consulted Limbach etc., Nucleic Acids Res. 22: 2183-2196,1994.As if described rare or uncommon RNA, the RNA (because they are generally the product of post transcriptional modificaiton) that is commonly referred to modification is to belong within the category of this paper term " RNA of unmodified ".The composition that the RNA of modification as herein described refers to nucleic acid promptly sugar, base and phosphoric acid part with naturally occurring, preferably with human body in the different molecule of naturally occurring composition.Although they are called modification " RNA ", owing to modify, so certainly comprise the molecule that is not RNA.The nucleosides surrogate is that ribose phosphoric acid skeleton be present in the molecule that the non-ribose phosphoric acid construct in the correct spatial relation is replaced by can be allowed base, thereby make that hybridization and the hybridization of using ribose phosphoric acid skeleton (for example, uncharged ribose phosphoric acid skeleton stand-in) are similar basically.This paper will discuss to above-mentioned example.

Modification as herein described can be introduced into double-stranded RNA arbitrarily as herein described and RNA sample molecule for example in the iRNA reagent.Can be preferably the antisense of iRNA reagent and in the sense strand one or two be all modified.Because nucleic acid is subunit or monomer polymerization body, thereby many modifications described below all appear on the position of repeating in the nucleic acid, and for example the disconnected property O to base or phosphoric acid position or phosphoric acid part modifies.In some cases, described modification is all body positions (subjectpostions) appearance at nucleic acid, but really not so under many (in fact most of) situation.For example, modification can occur over just 3 ' or 5 ' terminal position, can occur over just stub area, for example on the position of terminal nucleotide or chain last 2,3, modification can occur in double-stranded region in 4,5 or 10 Nucleotide, the strand zone takes place or in two zones.For example, the thiophosphatephosphorothioate of disconnected property O position is modified can only betide one or two end, can only betide stub area, for example on the position of terminal nucleotide or chain last 2,3,4, in 5 or 10 Nucleotide, perhaps can only betide strand and double-stranded region, especially terminal.Equally, modification can betide on sense strand, antisense strand or the two kinds of chains.In some cases, the adopted modification that has identical modification with antisense strand or have same type is arranged, but in other cases, there is justice just to have different modifications with antisense strand, for example, need in some cases only a chain to be modified, for example only sense strand is modified.

Introduce two main purposes of modifying are to have stability to degradation and improvement such as drug effect characteristic under biological environment pharmacological characteristics in iRNA reagent, these will further be discussed hereinafter.To the sugar in the iRNA reagent, base or skeleton carried out that other suitable being modified among the total PCT application No.PCT/US2004/01193 that was submitted on January 16th, 2004 is described to some extent.IRNA reagent can comprise the base that non-natural exists, for example the described base of submitting on April 16th, 2004 of total PCT application No.PCT/US2004/011822.IRNA reagent can comprise the sugar that non-natural exists, for example non-carbohydrate circular vectors molecule.The example characteristic that is used for the sugar that the non-natural of iRNA reagent exists is described in the total PCT application No.PCT/US2004/11829 that submitted on April 16th, 2003 to some extent.

IRNA reagent can comprise and connect (linkage) (for example the chirality thiophosphatephosphorothioate connects) between the Nucleotide that is used to increase the nuclease resistance.In addition, perhaps alternatively, iRNA reagent can comprise the ribose stand-in to increase the nuclease resistance.Be used for increasing and connect between the Exemplary core thuja acid of nuclease resistance and total PCT application No.PCT/US2004/07070 that the ribose stand-in were submitted on March 8th, 2004 describes to some extent.

IRNA reagent can comprise part-in conjunction with monomer subunit and suitable oligonucleotide synthetic monomer.The total U. S. application No.10/916 that exemplary monomer was submitted on August 10th, 2004 describes in 185 to some extent.

IRNA reagent can have the ZXY structure, and for example the total PCT application No.PCT/US2004/07070 that submits on March 8th, 2004 describes to some extent.

IRNA reagent can carry out compound with the both sexes composition.Exemplary both sexes composition used in the iRNA reagent is described in the total PCT application No.PCT/US2004/07070 that submitted on March 8th, 2004 to some extent.

In another embodiment, iRNA reagent can carry out compound with the delivery of agents with module mixture character.Described mixture comprise with following reagent in one or more (preferably two or more, more preferably three) support agent of linking to each other: (a) condensing agent (for example, can be as attracting the reagent as bind nucleic acid by ion or electrostatic interaction; (b) fusogen (for example, can merge and/or the reagent by cytolemma transportation; And (c) target group, cell or tissue targeting agent for example, as phytohemagglutinin, glycoprotein, fat or protein, as with specific cells type bonded antibody.With delivery of agents compound iRNA reagent description to some extent in the total PCT application No.PCT/US2004/07070 that was submitted on March 8th, 2004 mutually.

IRNA reagent can contain the non-standard pairing, for example having between justice and the antisense sequences at the iRNA duplex.The example feature of non-standard iRNA reagent is described in the total PCT application No.PCT/US2004/07070 that submitted on March 8th, 2004 to some extent.

Enhanced nuclease resistance

IRNA reagent such as the iRNA reagent of target influenza virus, can have enhanced nuclease resistance.

Increasing the total U. S. application No.60/559 that a method of resistance was submitted to as on May 4th, 2004, described in 917, is to identify cracking site and this site is modified to suppress cracking.For example, dinucleotides 5 ＇-ua-3 ＇, 5 ＇-ca-3 ＇, 5 ＇-ug-3 ＇, 5 ＇-uu-3 ＇ or 5 ＇-cc-3 ＇ can be used as cracking site.In some embodiments, all pyrimidines in the iRNA reagent all carry 2 ＇-modifications in sense strand, antisense strand or this two chain, so iRNA reagent has the enhanced resistance to endonuclease.Also can strengthen the nuclease resistance by 5 ' Nucleotide is modified, be 2 '-modified nucleotide thereby make the urea glycosides at least one 5 '-urea glycosides-VITAMIN B4-3 ' (5 ＇-ua-3 ＇) dinucleotides for example; 5 ＇-cytidine at least one 5 ＇-cytidine-VITAMIN B4-3 ＇ (5 ＇-ca-3 ＇) dinucleotides is 2 ＇-modified nucleotide; 5 ＇-urea glycosides at least one 5 ＇-urea glycosides-guanine-3 ＇ (5 ＇-ug-3 ＇) dinucleotides is 2 ＇ modified nucleotides; 5 ＇-urea glycosides at least one 5 ＇-urea glycosides-urea glycosides-3 ＇ (5 ＇-uu-3 ＇) dinucleotides is 2 ＇-modified nucleotide; Or the 5 ＇-cytidine at least one 5 ＇-cytidine-cytidine-3 ＇ (5 ＇-cc-3 ＇) dinucleotides is 2 ＇-modified nucleotide, described in International Application PCT/US2005/018931 of the co-applicant that submitted on May 27th, 2005.IRNA reagent can comprise at least 2, at least 3, at least 4 or at least 5 such dinucleotides.In particularly preferred embodiments, the Nucleotide of 5 ' Nucleotide in the 5 '-ua-3 ' and 5 ' that the occurs-ca-3 ' sequence motif of sense strand, antisense strand or these two kinds of chains for modifying.The 5 '-ua-3 ' that occurs of preferred sense strand, antisense strand or these two kinds of chains, the Nucleotide of 5 ' Nucleotide in 5 '-ca-3 ' and 5 '-ug-3 ' sequence motif for modifying.More preferably all pyrimidine nucleotides of sense strand are the Nucleotide of modification, and the Nucleotide of 5 ' Nucleotide in the 5 '-ua-3 ' and 5 ' that the occurs-ca-3 ' sequence motif of antisense strand for modifying, or have under the situation of 5 '-ug-3 ' sequence at all, antisense strand does not have 5 '-ua-3 ' and 5 '-ca-3 ' primitive.

Preferably, the Nucleotide of 2 '-modification comprises for example ribose unit of 2 '-modification.For example, 2 '-oh group (OH) can be modified or replace by multiple different " oxygen " or " deoxidation " substituting group.

The example that " oxygen "-2 ' oh group is modified comprises alkoxyl group or aryloxy (OR, for example R=H, alkyl, cycloalkyl, aryl, aralkyl, heteroaryl or sugar); Polyoxyethylene glycol (PEG); O (CH ₂CH ₂O) _nCH ₂CH ₂OR; " lock " nucleic acid (LNA), wherein 2 ＇ hydroxyls for example are connected with 4 ＇ carbon of same ribose by methylene bridge; O-amine (O-AMINE) and aminoalkoxy, O (CH ₂) _nAmine, (for example, amine=NH ₂, alkylamino, dialkyl amido, heterocyclic radical amino, virtue amino, ammonia diaryl base, heteroaryl amino or two heteroaryl aminos, diamines, polyamines).It should be noted that with the oligonucleotide that adopts strong thiophosphatephosphorothioate to modify and compare, only contain methoxyethyl group (MOE), (OCH ₂CH ₂OCH ₃, a kind of PEG derivative) oligonucleotide can show nuclease stability.

＂ deoxidation ＂ modifies and to comprise hydrogen (that is, ribodesose, the protuberance of itself and part ds RNA has special dependency); Halogen (for example fluorine); Ammonia (NH for example ₂, alkylamino, dialkyl amido, heterocyclic radical, virtue amino, ammonia diaryl base, heteroaryl amino, two heteroaryl aminos or amino acid); NH (CH ₂CH ₂NH) _nCH ₂CH ₂-amine (amine=NH ₂, alkylamino, dialkyl amido, heterocyclic radical amino, virtue amino, ammonia diaryl base, heteroaryl amino or two heteroaryl aminos) ,-NHC (O) R (R=alkyl, cycloalkyl, aryl, aralkyl, heteroaryl or sugar), cyano group, sulfydryl; Alkyl-sulphur-alkyl; Thio alkoxy; With alkyl, cycloalkyl, aryl, thiazolinyl and alkynyl, it can be replaced arbitrarily by for example amido functional group.

Preferred substituted is 2 ＇-methoxy ethyl, 2 ＇-OCH ₃, 2 ＇-O-allyl group, 2 ＇-C-allyl group and 2 ＇-fluorine.

In the oligonucleotide skeleton if comprise the cracking that furanose then also can reduce endonuclease.IRNA reagent can further be modified or modify by utilizing 3 ＇-3 ＇ to connect next nucleosides the conversion further at 3 ＇-ends by introducing 3 ' cation group.In other alternative, 3 ＇-ends are available to be sealed such as aminoalkyl groups such as 3 ＇ C5-aminoalkyl group dT.Other 3 ＇ binding substances also can suppress the cracking of 3 ＇-5 ＇ exonuclease.Be not subject to any theory, 3 ＇ binding substances such as Naproxen Base or Ibuprofen BP/EPs can combine with the 3 ＇-end of oligonucleotide by obstruction exonuclease on the space and suppress the cracking of exonuclease.Even the sugar of little alkyl chain, aromatic yl group or heterocycle binding substances or modification (D-ribose, ribodesose, glucose etc.) also can hinder 3 ＇-5 ＇-exonuclease.

Molten karyorhexis can be suppressed by introducing the modification of phosphoric acid ester linker, and for example phosphoric acid mercaptan ester connects.So preferred iRNA reagent comprises nucleotide dimer, this nucleotide dimer is rich in or is all modification phosphate-based of particular chiral form, this modification phosphate-based usually by oxygen occupied non-bridge location put and contain heteroatoms.Described heteroatoms can be S, Se, Nr ₂Or Br ₃When heteroatoms is S, preferably is rich in or is that chirality (chiral purity) Sp connects all.Be rich in and mean at least 70,80,90,95 or 99% preferred form.When being incorporated into 5 ＇ near iRNA reagent-or 3 ＇-end position, particularly 5 ＇-end position, the phosphoric acid ester linker of modification is effective especially for suppressing the exonuclease cracking.

5 ＇ binding substancess also can suppress the cracking of 5 ＇-3 ＇ exonuclease.Be not subject to any theory, can combine with the 5 ＇-end of oligonucleotide and suppress the cracking of exonuclease by hindering exonuclease on the space such as 5 ＇ binding substancess of Naproxen Base or Ibuprofen BP/EP etc.Even the sugar of little alkyl chain, aromatic yl group or heterocycle binding substances or modification (D-ribose, ribodesose, glucose etc.) also can hinder 3 ＇-5 ＇-exonuclease.

When the iRNA of two strandsization reagent comprised strand Nucleotide overhang at least one end, iRNA reagent can have the enhanced resistance to nuclease.In preferred embodiment, the Nucleotide overhang comprises 1-4, preferred 2-3 not paired Nucleotide.One preferred embodiment in, directly with terminal nucleotide to the strand overhang that is connected in pairs Nucleotide contain purine bases, and terminal nucleotide pair is that G-C is right, or 4 last complementary nucleotide centerings have and have at least two pairs to be that G-C is right.In further embodiment, the Nucleotide overhang can have 1 or 2 paired Nucleotide, and in an exemplary embodiment, described Nucleotide overhang is 5 ＇-gc-3 ＇.In preferred embodiment, the Nucleotide overhang is the 3 ＇-end at antisense strand.In one embodiment, iRNA reagent is included in the 5 ＇-cgc-3 ＇ motif of 3 ＇-ends of antisense strand, thereby forms 2-nt overhang 5 ＇-gc-3 ＇.

Thus, iRNA reagent can comprise in order to suppress the modification such as the degraded of nucleases such as endonuclease of finding or exonuclease in being tried the body body.This paper is called NRM or nuclease resistance with these monomers and promotes monomer, and will modify accordingly and be called NRM and modify.In many cases, these modify other characteristic that also can regulate iRNA reagent, for example with albumen such as translocator such as serum albumin or with the interactional ability of RISC member, perhaps first and second sequences form mutually duplex ability or with the ability that forms duplex such as another sequence of target molecule.

Can introduce one or more different NRM in iRNA reagent or in the sequence of iRNA reagent modifies.NRM modifies and can use more than 1 time in sequence or in iRNA reagent.

During modifying, NRM comprises that some only can be present in terminal modification and other can be present in the modification of any position.Some NRM modify and can suppress hybridization, therefore preferably it only are used for stub area, more preferably are not used in the cracking site or the cracking zone of (particularly in the antisense strand) in the sequence of target main body sequence or gene.They can be used for the optional position of sense strand, as long as two interchain hybridization of ds iRNA reagent can enough be kept.In some embodiments, preferably NRM is placed the cracking site or the cracking zone of sense strand, the silence of missing the target is minimized.

As a rule, it can be different distributions that NRM modifies, and this distribution is depended on that they are included in the sense strand and also is included in the antisense strand.If be included in the antisense strand, then not should with disturb or suppress the endonuclease cracked modify be inserted into the cracked zone of standing the RISC mediation as in cracking site or the cracking zone (as Elbashir etc., 2001, described in Genes and the Dev.15:188, it is incorporated herein by reference).The cracking of target occurs in 20 or the roughly mid-way of the antisense strand of 21nt, or occurs in first Nucleotide upstream about 10 or the 11 Nucleotide places of said target mrna (itself and antisense strand complementation).Cracking site as herein described refer to the cracking site both sides, with it hybridization the target thing or the Nucleotide on the iRNA reagent chain.The cracking zone refers to the Nucleotide in 1,2 of the cracking site any direction or 3 Nucleotide.

This modification can be introduced in the stub area, and the terminal position of justice or antisense strand or 2,3,4 or 5 positions are endways for example arranged.

Bolt is part (Tethered Ligands)

The character of iRNA reagent comprises its pharmacological property, for example can by introduce such as Bolt systemParts such as part influence and customize.In addition, the pharmacological property of iRNA reagent when no matter this iRNA reagent is or is exactly bolt when being part, is improved by introducing part.

Can all bolt be to iRNA reagent or as pharmaceutical adjunct or additive with multiple integrants such as parts, for example, bolt is to part-in conjunction with in the monomer subunit carrier.Described part-in conjunction with the example of monomer subunit hereinafter, but it only is preference, integrant also can be connected on the iRNA reagent in other position.

Preferred moiety is a part, and it connects preferably covalently directly or indirectly by intervention property key and is connected to carrier.In preferred embodiment, part is connected to carrier by intervention property key.When with part-when being introduced in the growing chain in conjunction with monomer, part or bolt are that part may reside in part-in conjunction with on the monomer.In some embodiments, after " precursor " part-in conjunction with the monomer subunit being introduced into growing chain, part can be introduced into " precursor " part-in conjunction with in the monomer subunit.For example, can for example monomer of N-terminal key, for example TAP-(CH will be had ₂) _nNH ₂, what be inserted into growth has in justice or the antisense strand.In operation subsequently, promptly, after being inserted into the precursor monomer subunit in this chain, make subsequently and have for example part of pentafluorophenyl esters or aldehyde radical of electrophilic group, by mode, with precursor ligands-be connected in conjunction with monomer with the electrophilic group of part and precursor ligands-be connected in conjunction with the terminal nucleophilic group of monomer subunit key.

In preferred embodiment, part changes distribution, target or the transformation period of its iRNA reagent of introducing.In preferred embodiment, with for example compare with the kind that does not have this part, part can provide the enhancing affinity at selected target compound, and described selected target compound for example is molecule, cell or cellular type, chamber such as cell chamber or organ chamber, tissue, organ or body part.

It is natural or modify few Yeast Nucleic Acid or contain monomer described herein and/or transhipment, hybridization and the specificity character of the poly molecule of the arbitrary combination of natural or modification property ribonucleotide and can improve its nuclease resistance that preferred part can improve gained.

Part generally includes and for example is used for sorbefacient therapeutic regulation thing; For example be used to monitor the diagnostic compounds or the reporter group of distribution; Linking agent; Give the composition of nuclease resistance; And natural or uncommon nucleic acid base.Common example comprises lipophilic molecules, lipid, plant agglutinin, steroid (for example Uvaol, agavogenin (hecigenin), diosgenin), terpenes (for example triterpene, as Sarsasapogenin, suberone, epifriedelinol deutero-lithocholic acid), VITAMIN, carbohydrate (for example dextran, pulullan polysaccharide, chitin, chitosan, synanthrin, cyclodextrin or hyaluronic acid), protein, protein binding agent, the plain target molecule of integration, polycation, peptide, polyamines and peptide analogs.

Described part can for example can be synthetic polymers such as synthetic polyamino acid for naturally occurring, reorganization or synthetic molecule.The example of polyamino acid comprises poly-lysine (PLL), poly L-aspartic acid, poly L-L-glutamic acid, styrene-maleic anhydride copolymer, poly (L-propiolactone-be total to-ethylene glycolization) multipolymer, divinyl ether-copolymer-maleic anhydride, N-(2-hydroxypropyl) methacrylamide copolymer (HMPA), polyoxyethylene glycol (PEG), polyvinyl alcohol (PVA), urethane, poly-(2-ethylacrylic acid), N-isopropyl acrylamide polyalcohol or poly-phosphine piperazine.The example of polyamines comprises: polymine, poly-lysine (PLL), spermine, spermidine, polyamines, false peptide polyamines, plan peptide class polyamines, dendrimer polyamines, arginine, amidine, protamine, cation constituent, cationization lipid for example, the quaternary ammonium salt of cationization porphyrin, polyamines or be alpha helical peptides.

Part also can comprise the target group, for example thyrotropin, melanotropin, surfactant protein A, mucoprotein sugar, glycosylation amino acids, Transferrins,iron complexes, bisphosphonate, polyglutamic acid salt, poly arginic acid salt or cell or tissue target agent such as RGD peptide or RGD peptide analogs.

Part can be albumen, and for example glycoprotein, low-density lipoprotein lipoprotein such as (LDL), human serum albumin albumin such as (HSA) or peptide are as having the molecule of special avidity, the antibody that combines with particular cell types such as cancer cells, endotheliocyte or osteocytes to being total to part.Part also can comprise hormone or hormone receptor.They also can comprise non-peptide class, for example cofactor, multivalence lactose, multivalence semi-lactosi, N-ethanoyl-GalN, N-ethanoyl-glucosamine, multivalence seminose or multivalence trehalose.Part can be for example lipopolysaccharides, p38 map kinase activator or the agent of NF-kB activation.

Described part can be a material such as medicine for example, and it can be by for example destroying the cytoskeleton of cell, for example increases the amount that absorbs the iRNA reagent to the cell by tubulin, microfilament and/or the Intermediate Filaments that destroys cell.Described medicine can be for example taxol (taxon), vincristine(VCR), vinealeucoblastine(VLB), cell fission chalone, R 17934, japlakinolide, latrunculin A (latrunculin A), phalloidin, swinholide A, indanocine or myoservin.

On the one hand, described part is a lipid or based on the molecule of lipid.This lipid or preferably combine with for example human serum albumin serum proteins such as (HSA) based on the molecule of lipid.The HAS binding partner allows binding substances to be distributed to target tissue such as hepatic tissue, comprises the parenchyma of liver.Other can also can be used as part with HAS bonded molecule.For example, can use neproxin or acetylsalicylic acid.Lipid or based on the molecule of lipid can: (a) increase the resistance of binding substances to degraded; (b) increase the target of target cell or cytolemma or increase transportation to target cell or cytolemma; And/or (c) can be used to regulate and combining such as serum proteins such as HAS.

Can be used to regulate based on the part of lipid and for example control combining of binding substances and target tissue.For example, be not easy that with the stronger lipid of the binding ability of HSA or based on the part of lipid kidney is had target, therefore be difficult for from body, being eliminated away.With a little less than the binding ability of HSA some lipid or can be used for the binding substances of targeting based on the part of lipid in kidney.

One preferred embodiment in, combine with HAS based on the part of lipid.Preferably, thus its with enough affinities combine with HAS make binding substances can preferred distribution to non-nephridial tissue.Yet, the degree that preferred described affinity can not be reversed to the combination of HAS-part by force.

On the other hand, described part is a kind of composition, for example is can be by the VITAMIN or the nutrition of target cell such as proliferative cell absorption.They are that the disease of feature is very useful for treatment with the cell proliferation of not expecting such as pernicious or non-mailgnant form such as cancer cells.The VITAMIN of example comprises vitamin A, E and K.Other exemplary VITAMIN comprises vitamin B group, and for example folic acid, B12, riboflavin, vitamin H, pyridoxal or other are by VITAMIN or nutrition that cancer cells absorbed.

On the other hand, described part is the Premeabilisation of cells agent, is preferably the agent of spirrillum Premeabilisation of cells.Preferred described reagent is amphoteric.Exemplary reagent is peptide such as Tat peptide or feeler foot peptide (antennapedia).If described reagent is peptide, then it can be modified, and comprises peptidyl stand-in, invertomer (invertomer), non-peptide or false peptide connector and uses D-amino acid.Spirrillum reagent is preferably α spiral reagent, its preferably have lipophilic mutually with thin fat mutually.

5 ＇-phosphoric acid ester is modified

In preferred embodiment, iRNA reagent be 5 ＇ phosphorylations or comprise the phosphoryl analogue at 5 ＇ initiating terminals.5 ＇ of antisense strand-phosphoric acid ester is modified the corresponding to modification of gene silencing that comprises with the RISC mediation.Suitable modification comprises: 5 ＇-phosplate ((HO) 2 (O) P-O-5 ＇); 5 ＇-biphosphonate ((HO) 2 (O) P-O-P (HO) (O)-O-5 ＇); 5 ＇-triguaiacyl phosphate ((HO) 2 (O) P-O-(HO) (O) P-O-P (HO) (O)-O-5 ＇); 5 ＇-guanosine cap (7-methylates or non-methylating) ((O) P-O-(HO) of 7m-G-O-5 ＇-(HO) (O) P-O-P (HO) (O)-O-5 ＇); 5 ＇-adenosine cap (Appp) and modify arbitrarily or the Nucleotide cap structure of unmodified ((O) P-O-(HO) of N-O-5 ＇-(HO) (O) P-O-P (HO) (O)-O-5 ＇); 5 ＇-single thiophosphate ester (phosphoric acid mercaptan ester; (HO) 2 (S) P-O-5 ＇); 5 ＇-single phosphorodithioate (di(2-ethylhexyl)phosphate mercaptan ester; (HO) (HS) 5 ＇-phosphoric acid mercaptan ester ((HO) 2 (O) P-S-5 ＇) (S) P-O-5 ＇); The phosplate that any oxygen/sulphur replaces, the extra combination of bisphosphate and triguaiacyl phosphate (e.g.5 ＇-α-thio triphosphates ester, 5 ＇-γ-thio triphosphates ester etc.), 5 ＇-phosphoramidate ((HO) 2 (O) P-NH-5 ＇, (HO) (NH2) (O) P-O-5 ＇), 5 ＇-alkyl phosphonates (R=alkyl=methyl, ethyl, sec.-propyl, propyl group etc., for example RP (OH) (O)-O-5 ＇-, (OH) 2 (O) P-5 ＇-CH2-), 5 ＇-alkyl ether phosphate (R=alkyl oxide=methoxymethyl (MeOCH2-), ethoxyl methyl, Deng, for example RP (OH) (O)-O-5 ＇-).

Can modify so that thereby the sense strand inactivation suppresses the formation of active RISC sense strand, come to reduce potentially the effect of missing the target thus.This can be by the 5 ＇-phosphorylation that prevents sense strand, as finishing by modifying (referring to Nykanen etc. with 5 ＇-O-methyl ribonucleotides, (2001) ATP requirementsand small interfering RNA structure in the RNA interference pathway.Cell 107,309-321.).Also can use other modification that prevents phosphorylation, for example, only replace 5 ＇-OH with H rather than O-Me.Perhaps, thus can add a bulky group to 5 ＇-phosphoric acid ester be translated into the phosphodiester connector.

The non-natural base

Nitro-pyrrole base and nitroindoline base are the class non-natural bases in known universal base (universal bases) compound.Universal base is can replace any 4 naturally occurring bases and habit of unwinding (melting behavior) and active compound that can substantial effect oligonucleotide duplex.Opposite with stabilization, hydrogen bond action and naturally occurring base have relation, suppose that the oligonucleotide duplex that contains 3-nitro-pyrrole base base only stablizes by accumulation.There is not significant hydrogen bond action to eliminate the specificity of particular complementary base with nitro-pyrrole base base.In addition, various reports confirm 4-, and 5-and 6-nitroindoline base are very little for the specificity of four natural bases.Ironically, the oligonucleotide duplex that contains 5-nitroindoline base is more stable than the oligonucleotide that contains 4-nitroindoline base and 6-nitroindoline base accordingly.The preparation 1-(2 '-O-methyl-β-D-ribofuranosyl)-5-nitroindoline process at Gaubert, G.; Wengel has explanation among the J.Tetrahedron Letters 2004,45,5629.Other universal base of available of the present invention comprise the xanthoglobulin base, different xanthoglobulin glycosides, 2-azepine-xanthoglobulin glycosides, 7-denitrification-xanthoglobulin glycosides, the nitroimidazole base, nitropyrazole base, nitrobenzimidazole base, nitro indazole base, the amino indole base, pyrroles's pyrimidyl, and structural derivative.For nitro-pyrrole base, nitroindoline base and other above-mentioned universal base of mentioning are discussed more specifically, comprise synthesis step, see also Vallone etc., Nucleic AcidsResearch, 27 (17): 3589-3596 (1999); Loakes etc., J.Mol.Bio., 270:426-436 (1997); Loakes etc., Nucleic Acids Research, 22 (20): 4039-4043 (1994); Oliver etc., OrganicLetters, Vol.3 (13): 1977-1980 (2001); Amosova etc., Nucleic Acids Research, 25 (10): 1930-1934 (1997); Loakes etc., Nucleic Acids Research, 29 (12): 2437-2447 (2001); Bergstrom etc., J.Am.Chem.Soc., 117:1201-1209 (1995); Franchetti etc., Biorg.Med.Chem.Lett.11:67-69 (2001); With Nair etc., Nucelosides, Nucleotides ﹠amp; Nucleic Acids, 20 (4-7): 735-738 (2001).

The difluoro toluene base is the non-natural base as universal base.The difluoro toluene base is the isostere of natural base thymus pyrimidine.Unlike thymus pyrimidine, the difluoro toluene base does not demonstrate the selectivity to any natural base.Can be used for other aromatic compounds as universal base of the present invention is 4-fluoro-6-tolimidazole and 4-tolimidazole.In addition, relevant hydrophobicity isoquinolone derivatives 3-methyl isoquinolone base, 5-methyl isoquinolone base is to cause the oligonucleotide duplex to compare unsettled slightly universal base with the oligonucleotide sequence that only contains natural base with 3-methyl-7-proyl isoquinolone.Other non-natural bases of the present invention's expection comprise the 7-azaindolyl, 6-methyl-7-azaindolyl, imidazole pyridyl, 9-methyl-imidazole pyridyl, pyrroles's pyrazinyl, isoquinolone, 7-proyl isoquinolone, proyl-7-azaindolyl, 2,4-skatole base, 4,6-dimethylated indolyl, phenyl, naphthyl, anthryl, benzene anthryl, pyrenyl, diphenylethyllene, naphthacenyl, pentacenyl, and structural derivative.For the difluoro toluene base is discussed more specifically, 4-fluoro-6-tolimidazole, 4-tolimidazole and above-mentioned other non-natural bases comprise synthesis step, see also Schweitzer etc., J.Org.Chem., 59:7238-7242 (1994); Berger etc., Nucleic Acids Research, 28 (15): 2911-2914 (2000); Moran etc., J.Am.Chem.Soc., 119:2056-2057 (1997); Morales etc., J.Am.Chem.Soc., 121:2323-2324 (1999); Guckian etc., J.Am.Chem.Soc., 118:8182-8183 (1996); Morales etc., J.Am.Chem.Soc., 122 (6): 1001-1007 (2000); McMinn etc., J.Am.Chem.Soc., 121:11585-11586 (1999); Guckian etc., J.Org.Chem., 63:9652-9656 (1998); Moran etc., Proc.Natl.Acad.Sci., 94:10506-10511 (1997); Das etc., J.Chem.Soc., PerkinTrans., 1:197-206 (2002); Shibata etc., J.Chem.Soc., Perkin Trans., 1:1605-1611 (2001); Wu etc., J.Am.Chem.Soc., 122 (32): 7621-7632 (2000); O ' Neill etc., J.Org.Chem., 67:5869-5875 (2002); Chaudhuri etc., J.Am.Chem.Soc., 117:10434-10442 (1995); With U.S. patent 6,218,108.

Provide among the PCT/US2005/025967 of further details pending trial in the same applicant who submitted on July 21st, 2005 of the synthetic and application of universal base, incorporate its full content into by reference here.

Particularly useful under the situation of the gene of universal base in attempting the target life entity, this gene has shown variability between the different strains of this life entity.In the viral genome zone that is considered to high conservative usually is such.Introduce universal base and can design the iRNA reagent of a large amount of different virus strains of target, even this virus stain is different on one or several (for example maximum 3) nucleotide position.

Consider the specificity and the activity of iRNA reagent, universal base preferably is included in iRNA reagent in the least responsive zone of Nucleotide mispairing.The 2-9 position of antisense strand that has shown iRNA reagent is the most responsive for the mispairing between antisense strand and said target mrna, and " seed region " that this zone is called iRNA reagent (seed-region).Therefore, when one or more universal base were introduced iRNA reagent, it preferably was incorporated into outside the seed region.

The complimentary positions that table 1F and table 1G are presented at sense strand and antisense strand comprises the iRNA reagent of universal base.But, though this be one of iRNA reagent of the present invention preferred embodiment, when universal base was present in the antisense strand, the effect of the universal base in the iRNA reagent can be more obvious.Therefore can envision in sense strand with antisense strand in the locational base of universal base paired can be universal base or any other suitable base, for example a, u, c or g.Preferably, the people's base that can test which position in sense strand can give iRNA reagent the highest activity and/or selectivity.Perhaps, be chosen in the base on this specific position of most target gene variants, stipulate that described target gene variant is subjected to the inhibition of the iRNA that discussed in expression.

IRNA reagent transports into cell

Do not expect to be bound by any theory, can cause combining of iRNA reagent and lipoprotein (as LDL, HDL) in the blood in conjunction with the iRNA reagent of cholesterol and the chemical similarity between some lipoprotein composition (as cholesterol, cholesteryl ester, phosphatide), and/or cause iRNA reagent with to cholesterol, the cellular constituent that has avidity as the composition in the cholesterol transport pathway interacts.Lipoprotein with and moiety absorbed by cell by various actives or passive transport mechanism or process, as without restriction, the endocytosis of the LDL of LDL-receptors bind, by with the endocytosis of the interactional oxidisability of Scavenger acceptor A or other modification property LDL, HDL cholesterol absorption in the liver of Scavenger acceptor B1-mediation, the cholesterol of ABC (ATP is in conjunction with the box) translocator of pinosome or utilization such as ABC-A1, ABC-G1 or ABC-G4 is striden the film transportation.So, help having the absorption of the cell of described transport mechanism such as liver cell to it in conjunction with the iRNA reagent of cholesterol.So, the invention provides evidence and method in common and iRNA reagent is targeted to the cell of expressing some cell surface composition such as acceptor, it is to be attached to iRNA reagent by the native ligand with described composition (as cholesterol), or by a kind of chemical substance (as cholesterol) is attached to be combined with as described on the iRNA reagent of native ligand of composition (for example LDL, HDL).

Other embodiment

RNA as iRNA reagent, can produce in cell paste, for example by the foreign DNA template is transported into cell.For example, dna profiling can be inserted into carrier and as gene therapy vector.Gene therapy vector can be delivered to the experimenter by intravenous injection, topical application (U.S. patent No.5,328,470) or stereotactic injection (referring to Chen etc., Proc.Natl.Acad.Sci.USA 91:3054-3057,1994).The pharmaceutical preparation of gene therapy vector is included in the gene therapy vector in the acceptable diluent, or comprises a kind of sustained-release matrix, and wherein gene delivery carrier is embedded in this sustained-release matrix.Dna profiling for example comprises two transcription units, and a generation comprises the transcript of iRNA reagent upstream chain, and another generation comprises the transcript of iRNA reagent downstream chain.When template was transcribed, iRNA reagent generated and processedly advances in the siRNA reagent fragment this fragment mediated gene silencing.

Preparation

The present invention also comprises pharmaceutical composition and the preparation that contains dsRNA compound of the present invention.Pharmaceutical composition of the present invention can be as required local (local) still whole body therapeutic and use in many ways according to zone to be treated.To use can be external application (topical), through lung as by sucking or spraying powder or aerosol, comprise the employing sprays; In respiratory tract, nose, epidermis and corium, oral cavity or parenteral.Parenteral administration comprises intravenously, intra-arterial, subcutaneous, intraperitoneal or intramuscular injection or infusion; Or use in the skull, for example in the sheath or in the ventricle.

The pharmaceutical composition of external application and preparation can comprise transdermal subsides, ointment, washing lotion, frost, gel, drops, suppository, sprays, liquid or powder.Need conventional pharmaceutical carrier, water, powder or oil base, thickening material etc.The condom that applies, gloves etc. are also useful.Preferred external preparation comprises that those make dsRNAs of the present invention and external application delivery agents blended preparation mutually, and described external application delivery agents for example is lipid, liposome, lipid acid, fatty acid ester, steroid, sequestrant and tensio-active agent.Preferred lipid and liposome comprise neutral (DOPE=DOPE for example, dimyristoyl phosphatidyl choline=DMPC, distearoyl phosphatidylcholine) (for example GLYCEROL,DIMYRISTOYL PHOSPHATIDYL=DMPG) and cationic (for example two oleoyl tetramethyl-aminopropyl=DOTAP and the DOPE=DOTMA) of negative electricity.DsRNAs of the present invention can be enclosed in the liposome or with its formation mixture, particularly cationic-liposome.Perhaps, dsRNAs can be compound with lipid, particularly cation lipid.Preferred lipid acid and ester include but not limited to arachidonic acid, oleic acid, eicosanoic acid, lauric acid; sad, capric acid, tetradecanoic acid, palmitinic acid; stearic acid, linolic acid, linolenic acid; dicaprate, three decylates, XU 61518.10; GLYCERYL DILAURATE, the single caprin of 1-, 1-dodecyl-aza-cycloheptane-2-ketone; fatty acyl carnitine, acyl group choline, or C _1-10Alkyl ester (for example isopropyl myristic acid ester IPM), monoglyceride, triglyceride or its pharmacologically acceptable salts.The U.S. patent application serial number .09/315 that external preparation was submitted on May 20th, 1999 specifies in 298, here by with reference to quoting its full content.

Oral compositions and preparation comprise suspension in powder or particle, particulate, nanoparticle, water or the non-aqueous media or solution, capsule, gel capsule, pouch, tablet or mini.Thickening material can be arranged, seasonings, thinner, emulsifying agent, dispersing auxiliary or tackiness agent.Preferred oral preparations is to make dsRNAs of the present invention and those co-administered preparations of one or more penetration enhancers, tensio-active agent and sequestrant.Preferred surfactants comprises lipid acid and/or its ester or salt, bile acide and/or its salt.Preferred bile acide/salt comprises gallodesoxycholic acid (CDCA) and Ursodeoxycholic Acid (UDCA) (UDCA), cholic acid, Felacrinos, Septochol; paddy ammonia cholic acid, glycocholic acid, sugared Septochol, taurocholate; tauroursodeoxycholic acid, ox sulphur-24,25-dihydro-Sodium Fusidate and sugared dihydro Sodium Fusidate.Preferred lipid acid comprises arachidonic acid, undecanoic acid, oleic acid, lauric acid; sad, capric acid, tetradecanoic acid, palmitinic acid; stearic acid, linolic acid, linolenic acid, dicaprate; three decylates, XU 61518.10, GLYCERYL DILAURATE; the single caprin of 1-, 1-dodecyl-aza-cycloheptane-2-ketone, fatty acyl carnitine; acyl group choline, or monoglyceride, triglyceride or its pharmacologically acceptable salts (for example sodium salt).The preferably combination of penetration enhancers, for example combination of fatty acid/salt and bile acide/salt.Particularly preferred combination is the sodium salt of lauric acid, capric acid and UDCA.Other penetration enhancers comprises polyoxyethylene-9-lauryl, polyoxyethylene-20-cetyl ester.DsRNAs of the present invention can be with particle form that comprises spray-dried granules or the form oral delivery that is compounded to form micron or nano particle.The DsRNA complexing agent comprises poly--amino acid; Poly-imines; Polyacrylic ester; Polyalkyl acrylate, polyoxy ethane,, Polyalkylcyanoacrylanano; Cationization gelatin, albumin, starch, acrylate, polyoxyethylene glycol (PEG) and starch; Polyalkyl alpha-cyanacrylate; The DEAE-deutero-gathers imines, short stalk mould polysaccharide (pollulans), Mierocrystalline cellulose and starch.Particularly preferred complexing agent comprises chitosan, N-trimethyl chitin, poly-L-Lysine, polyhistidyl, poly ornithine, poly-spermine, protamine, the polyvinyl pyrimidine, poly-sulfo-diethylamino methyl ethene P (TDAE), poly-amino-benzene ethene (for example p-amino), poly-(Methyl 2-cyanoacrylate), poly-(cyanacrylate), poly-(Tisuacryl), poly-(isobutylcyanoacrylate), poly-(alpha-cyanoacrylate dissident ester), the DEAE-methacrylic ester, DEAE-Ethyl acrylate, DEAE-acrylamide, DEAE-albumin and DEAE-dextran, polymethyl acrylate, the own ester of polyacrylic acid, poly-(D, L-lactic acid), poly-(DL-lactic acid-co-oxyacetic acid (PLGA), alginate, and polyoxyethylene glycol (PEG).The oral preparations of dsRNAs and preparation thereof are at U.S. patent application Ser.No.08/886,829 (submissions on July 1st, 1997), Ser.No.09/108,673 (submissions on July 1st, 1998), Ser.No.09/256,515 (submissions on February 23rd, 1999), Ser.No.09/082,624 (submission on May 21st, 1998) and Ser.No.09/315 have detailed description in 298 (submissions on May 20th, 1999), quote its full content by reference here.

Parenteral, sheath is interior or ventricle interior (intraventricular) is used composition and preparation can comprise the aseptic aqueous solution that contains buffer reagent, thinner or other suitable additives, described additive is such as but not limited to penetration enhancers, carrier compound and other pharmaceutically acceptable carriers or vehicle.

Pharmaceutical composition of the present invention includes but not limited to solution, emulsion and contains the preparation of liposome.These compositions can be generated by various components, and these components include but not limited to prefabricated liquid, self-emulsification solid and self-emulsifying semisolid.

The conventional pharmaceutical formulations of the present invention that exists with unit dosage form can prepare according to known routine techniques in the pharmaceutical industry.These technology comprise activeconstituents and pharmaceutical carriers or vehicle bonded step.Usually, preparation prepares by the solid carrier of activeconstituents and liquid vehicle or segmentation or both evenly and are closely combined, and then, if desired, product is formalized.

Composition of the present invention can be mixed with any possible formulation, such as but not limited to tablet, capsule, gel capsule, syrup, soft gel, suppository and enema.Composition of the present invention can also be mixed with suspension in water-based, non-water or blending agent.Waterborne suspension can also contain the material that increases suspension viscosity, for example comprises Xylo-Mucine, sorbyl alcohol and/or dextran.Suspension can also contain stablizer.

In an embodiment of the invention, pharmaceutical composition can be mixed with or use with form of foam.Medicinal foam comprise formulation example as, but be not limited to emulsion, micro emulsion, frost, gelifying agent and liposome.Although similar substantially in essence, these preparations are variant on the composition of the finished product and consistence.The such composition and the preparation of preparation are known for the technician of pharmacy and formulation art, and can be used for the preparation of composition of the present invention.

Emulsion

Composition of the present invention can prepare or be mixed with emulsion.Emulsion be a kind of liquid in another kind with the drop dispersive heterogeneous system (Idson of diameter greater than 0.1 μ m, in Pharmaceutical DosageForms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., the 1st volume, the 199th page; Rosoff, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., the 1st volume, the 245th page; Block in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., the 2nd volume, the 335th page; Higuchi etc., in Remington ＇ s Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa., 1985, the 301 pages).Emulsion usually is the binary system that comprises close mixing and mutual immiscible two liquid phases of dispersive.Generally, emulsion can be water-in-oil (w/o) or oil-in-water (o/w) type.When water by fine and closely woven separately and when being dispersed in a large amount of oil phases with droplet, resulting composition is called water-in-oil (w/o) emulsion.Perhaps, when oil by fine and closely woven when being dispersed in a large amount of aqueous phase separately and with droplet, resulting composition is called oil-in-water (o/w) emulsion.Emulsion can also comprise other components and active medicine except disperse phase, it can be at water, oil phase or this is as the solution that separates phase.As required, also can there be pharmaceutical excipient in emulsion, for example emulsifying agent, stablizer, dyestuff and antioxidant.The pharmacy emulsion can also be a multiple emulsion, and it comprises greater than two-phase, for example under the situation of oil-water-oil (o/w/o) and water-oil-water (w/o/w) emulsion.The compound preparation usually has the advantage that simple binary emulsion does not have like this.The independent oil droplet of o/w emulsion constitutes the w/o/w emulsion round little water droplet in the multiple emulsion.The system that comprises oil droplet in the same globule stable in oils external phase is the o/w/o emulsion.

Emulsion is characterised in that thermodynamic instability.Usually, the disperse phase of emulsion or discontinuous phase be dispersed in well outside mutually or in the external phase and the viscosity by emulsifying agent or preparation keep this form.Emulsion can be semi-solid state or solid-state mutually, as emulsion ointment or breast frost.The method of other stable emulsions need be used emulsifying agent, and it can add any phase of emulsion.Emulsifying agent can extensively be divided into 4 classes: synthetic tensio-active agent, naturally occurring emulsifying agent, absorption base and finely divided solid (Idson, in PharmaceuticalDosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., the 1st volume, the 199th page).

The synthetic tensio-active agent is also referred to as tensio-active agent, in emulsion formulations, is widely used, and (the Rieger that in the literature comment arranged, in Pharmaceutical Dosage Forms, Lieberman, Rieger andBanker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume1, p.285; Idson, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), Marcel Dekker, Inc., New York, N.Y., 1988, volume 1, p.199).Tensio-active agent is generally amphoteric, and comprises hydrophilic and hydrophobic part.Tensio-active agent is hydrophilic to be defined as hydrophilic (HLB) with ratio hydrophobic property, and it is the effective tool that tensio-active agent is classified and selected in the preparation preparation.Tensio-active agent can be divided into different types according to the character of hydrophilic radical: non-ionic type, anionic, cationic and both sexes (Rieger, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., the 1st volume, the 285th page).

Naturally occurring emulsifying agent used in the emulsion formulations comprises lanolin, beeswax, phosphatide, Yelkin TTS and Sudan Gum-arabic.The absorption base possess hydrophilic property, but it can absorb water and form the w/o emulsion keep semi-solid denseness, for example lanolin anhydrous bp93 and hydrophilic petrolatum.Also can be with the solid of segmentation as the good emulsifying agent, particularly and in the heavy-gravity preparation time with combinations-of surfactants.These solids comprise the polarity inoganic solids, heavy metal hydroxide for example, and unexpansive pottery is wilkinite for example, attapulgite, hectorite, kaolin, montmorillonite, colloidal silicic acid aluminium and colloidal silicic acid magnalium, pigment and non-polar solid be carbon or Tristearoylglycerol for example.

Also comprise miscellaneous non-emulsified material in the emulsion formulations, it exists for emulsion property and makes contributions.These non-emulsified materials comprise fat, oil, wax, lipid acid, Fatty Alcohol(C12-C14 and C12-C18), fatty acid ester, wetting agent, hydrophilic colloid, sanitas and antioxidant (Block, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., the 1st volume, the 335th page; Idson, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., the 1st volume, the 199th page).

Hydrophilic colloid or hydrocolloid comprise for example saccharan (for example, Sudan Gum-arabic, agar of natural rubber and synthetic polymkeric substance, alginic acid, carrageenin, guar gum, kuteera gum, and tragacanth gum), derivatived cellulose (for example, carboxymethyl cellulose and carboxy-propyl cellulose), and synthetic polymer (for example, carbomer, ether of cellulose, and carboxyl ethylene polymer).These disperse or expand to form colloidal solution in water, and it is by forming the strong interface film and the stable emulsion by the viscosity that increases outside phase around disperse phase liquid.

Because emulsion often contains the composition of a large amount of support microorganism growth, for example carbohydrate, albumen, sterol and phosphatide, these preparations usually comprise sanitas.Sanitas commonly used in the emulsion formulations comprises Tegosept M, propylben, quaternary ammonium salt, benzalkonium chloride, p-hydroxybenzoate and boric acid.It is rotten to prevent preparation that emulsion formulations also usually adds antioxidant.Used antioxidant can be a radical scavenger, for example tocopherol, alkyl gallates, butylated hydroxy anisole (BHA), butylhydroxy toluene or reductive agent, for example citric acid, tartrate and Yelkin TTS of xitix and Sodium Pyrosulfite and antioxidant synergist for example.

Emulsion formulations is by the application and the existing in the literature (Idson that comments of manufacture method thereof in skin, oral cavity and parenteral path, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., the 1st volume, the 199th page).The emulsion formulations of oral delivery is because but preparation is convenient and the efficient height of absorption and biology availability, thereby (Rosoff, in Pharmaceutical Dosage Forms, Lieberman have been widely used, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p.245; Idson, inPharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, MarcelDekker, Inc., New York, N.Y., volume1, p.199).Mineral oil based laxative, oil-soluble vitamins and high fat nutritional formulation also belong to the oral material commonly used of o/w emulsion.

In an embodiment of the invention, the composition of dsRNAs and nucleic acid is formulated into micro emulsion.Micro emulsion is defined as the system of water, oil and amphipath, it is independent vision homogeneous and thermodynamically stable liquor (Rosoff, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., the 1st volume, the 245th page).Usually, micro emulsion is by elder generation oil to be distributed in the aqueous tenside solution, and the 4th component (the normally alcohol of medium chain) that adds q.s then prepares to form transparent system.Therefore, micro emulsion is thermodynamically stable isotropic clarification dispersion of two kinds of immiscible liquid, its interfacial film by surface active molecules is stablized (Leung and Shah, in:Controlled Release of Drugs:Polymers andAggregate Systems, Rosoff, M., Ed., 1989, VCH Publishers, New York, the 185-215 page or leaf).Micro emulsion often prepares by making up 3～5 kinds of components, comprises oil, water, tensio-active agent, cosurfactant and ionogen.Micro emulsion is the character that water-in-oil (w/o) or oil-in-water (o/w) depend on oil and used tensio-active agent, also depend on the polar head of surfactant molecule and structure and how much filling (geometric packing) (Schott of hydro carbons tail, in Remington ' s Pharmaceutical Sciences, MackPublishing Co., Easton, Pa., 1985, p.271).

Broad research utilize the phenomenological method of phasor, and those skilled in the art have obtained comprehensive knowledge (Rosoff, in Pharmaceutical Dosage Forms, Lieberman to how preparing micro emulsion, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p.245; Block, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p.335).Compare with conventional emulsions, the advantage of micro emulsion is the water-fast medicine of dissolving in the thermodynamically stable drop preparation that forms naturally.

The used tensio-active agent of preparation micro emulsion comprises but is not limited to ionic surface active agent, nonionogenic tenside, Brij96, polyoxyethylene oleyl ether, polyglycerol fatty acid glyceryl ester, mono laurate four glyceryl ester (ML310), single oleic acid four glyceryl ester (MO310), single oleic acid six glyceryl ester (PO310), five oleic acid, six glyceryl ester (PO500), single capric acid ten glyceryl ester (MCA750), single oleic acid ten glyceryl ester (MO750), half oleic acid, ten glyceryl ester (SO750), ten oleic acid, ten glyceryl ester (DAO750) make up separately or with cosurfactant.Cosurfactant is generally short chain alcohol, for example ethanol, 1-propyl alcohol and 1-butanols, and it is used for producing abnormal film by being penetrated into the tensio-active agent film and forming the space between surfactant molecule, thereby increases the interface flowability.But micro emulsion can prepare without cosurfactant, and no pure self-emulsifying micro-emulsion system known in the art.Water is general but to be not limited to be the derivative of the aqueous solution, glycerine, PEG300, PEG400, Polyglycerine, propylene glycol and ethylene glycol of water, medicine.Oil phase includes but not limited to for example Captex 300, Captex 355, Capmul MCM, fatty acid ester, medium chain (C ₈-C ₁₂) monoglyceride, triglyceride and triglyceride level, polyoxyethylated fatty acid ester, Fatty Alcohol(C12-C14 and C12-C18), the glyceryl ester of Pegylation, the glyceryl ester of saturated polyglycolysed, the C of saturated polyglycolysed ₈-C ₁₀Glyceryl ester, vegetables oil and silicone oil.

Consider that from drug solutionization and enhancing drug absorption angle micro emulsion is subjected to special concern.Lipid base micro emulsion (o/w and w/o) has been proposed to strengthen bioavailability of medicament, comprises peptide (Constantinides etc., Pharamaceutical Research, 1994,11,1385-1390; Ritschel, Meth.Find.Exp.Clin.Pharmacol., 1993,13,205).Micro emulsion has the drug solutionization of improvement, prevents the medicine enzymolysis, may since tensio-active agent inductive membrane fluidity strengthen absorption with infiltrative variation, be convenient to prepare, compare with solid dosage be convenient to Orally administered, improve clinical efficacy and reduce toxic advantage (Constantinides etc., Pharamaceutical Research, 1994,11,1385; Ho etc., J.Pharm.Sci., 1996,85,138-143).When at room temperature mixing their component, the usually spontaneous formation of micro emulsion.This is particularly advantageous when preparation heat labile medicine, peptide or dsRNAs.Micro emulsion also has effectiveness active ingredient transdermal delivery when makeup or pharmaceutical application.Expect that micro emulsion composition of the present invention and preparation help increasing dsRNAs and nucleic acid and absorb from the gi tract whole body, and improve dsRNAs and nucleic acid and use local cells picked-up in the zone at gi tract, vagina, oral cavity and other.

Micro emulsion of the present invention can contain other component and additive ((Grill3) for example, caprylic/capric polyethylene glycol glycerol ester (Labrasol), and penetration enhancers), with the character of improving preparation and the absorption that strengthens dsRNAs of the present invention and nucleic acid.Penetration enhancers in the micro emulsion of the present invention can be divided into the class in the 5 big classes: tensio-active agent, lipid acid, biliary salts, sequestrant and non-chelating nonsurfactant (Lee etc., Critical Reviews in Therapeutic Drug Carriers Systems, 1991, p.92).Each class of having discussed above.

Liposome

Except being studied and being used for the micro emulsion of pharmaceutical preparation, also have the surfactant structures of a lot of rules, comprise individual layer, micella, bilayer and vesicle.Vesicle is liposome for example, considers from the aspect that medicine is sent, owing to its specificity and the lasting extensive concern that is subjected to of effect.Term used herein ＂ liposome ＂ refers to by being arranged as the vesicle that spherical bilayer or double-deck amphoteric lipid are formed.

Liposome is single sheet or splintery vesicle, and it has the film that is formed by lipophilic material and water kernel.Aqueous portion contains the composition that remains to be sent.Cationic-liposome has the advantage that can merge with cell walls.Though the non-cationic liposome can not effectively merge with cell walls, can be absorbed in vivo by scavenger cell.

In order to pass complete mammalian skin, lipid vesicles must be passed a series of pores, and each diameter is subjected to the suitable influence of wearing the skin gradient less than 50nm.Therefore, need using can high deformation and pass the liposome of such pore.

Other advantage of liposome comprises: the liposome physiologically acceptable and the biodegradable that are obtained by natural phospholipid; Liposome can add a large amount of water and the soluble medicine of lipid; Liposome can prevent that medicine that portion within it seals is by metabolism and degraded (Rosoff, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p.245).What need that emphasis considers at the preparation Liposomal formulation is the water-based volume of lipid surface charge, vesicle size and liposome.

Liposome is for the active substance transmission with to be delivered to action site very useful.Because liposome membrane is similar to biofilm structure, when liposome was used to organize, liposome began to merge with cytolemma, and along with liposome merges, cell is grown up, and the liposome content enters cell, and activeconstituents is had an effect there.

Liposomal formulation is owing to the focus that model becomes broad research of sending of multiple medicine.More and more evidences shows that for external application liposome has been compared multiple advantage with other preparations.Such advantage comprises that the high whole body that reduces with using absorbs relevant side effect, increases the accumulation of using on the target, and multiple medicine (hydrophilic or hydrophobic) can be applied skin.

The explanation that several pieces of reports are detailed the effect of liposome to delivery of agents, comprise polymer DNA is delivered in the skin.Compound (comprising analgesic agent, antibody, hormone and polymer DNAs) can be delivered in the skin.It is target that majority is used with the upper epidermis.

Liposome is divided into two big classes.Cationic-liposome is the liposome of positively charged, and itself and electronegative dna molecular interact and form stabilized complex.The DNA/ liposome complex and the electronegative cell surface of positively charged are combined in internalization in the endosome.Because the acid pH in the endosome, liposome breaks, content is discharged in the tenuigenin (Wang et al., Biochem.Biophys.Res.Commun., 1987,147,980-985).

Liposome is that pH-is responsive or electronegative, catches DNA rather than mixture.Because DNA is charged similar with lipid, therefore repulsion rather than compound can take place.But some DNA is hunted down in the water-based kernel of liposome.PH-sensitive lipid body has been used for the dna encoding thymidine kinase gene is delivered in the cell monolayer of cultivation.Expression of exogenous gene detected by target cell (Zhou etc., Journal ofControlled Release, 1992,19,269-274).

A kind of liposome composition of main type comprises the phosphatide except natural deutero-phosphatidylcholine.Neutral fat plastid composition can by, for example dimyristoyl phosphatidyl choline (DMPC) or dipalmitoyl phosphatidylcholine (DPPC) form.The cationic-liposome composition is generally formed by GLYCEROL,DIMYRISTOYL PHOSPHATIDYL, and the liposome of cationic membrane fusion is simultaneously mainly formed by DOPE (DOPE).Another kind of liposome composition is formed by phosphatidylcholine (PC), for example, and soybean PC and ovum PC.Another kind of then be that mixture by phosphatide and/or phosphatidylcholine and/or cholesterol forms.

The local delivery of liposomal pharmaceutical preparation to skin estimated in some researchs.The application that contains the liposome of the Interferon, rabbit of guinea pig skin causes skin bleb to reduce, and send Interferon, rabbit (for example as solution or emulsion) by other modes is invalid (Weiner etc., Journal of Drug Targeting, 1992,2,405-410).And then, also other research tested Interferon, rabbit as the part of Liposomal formulation with respect to using water-based system to send the usefulness of Interferon, rabbit, and sum up Liposomal formulation and be better than water-based system (duPlessis etc., Antiviral Research, 1992,18,259-265).

Effectiveness when the nonionic liposomal systems has been verified on delivering drugs into skin particularly comprises the system of nonionogenic tenside and cholesterol.The nonionic Liposomal formulation comprises Novasome.TM.I (GLYCERYL DILAURATE/cholesterol/polyoxyethylene-10-stearyl ester) and Novasome.TM.II (distearin/cholesterol/polyoxyethylene-10-stearyl ester), and it is used for S-Neoral-A is delivered to the corium of mouse.The result shows that such nonionic liposomal systems effectively promotes the different layers (S.T.P.Pharma.Sci. such as Hu, 1994,4,6,466) that S-Neoral-A deposits to skin.

Liposome also comprises the ＂ liposome of ＂ spatial stability, and this term refers to comprise the liposome of one or more specificity lipids here, when joining liposome, compares with the liposome that lacks such specificity lipid, has strengthened cycle life.The example of the liposome of spatial stability is the part in the lipid part of the vesicle formation of liposome, (A) comprises one or more glycolipids, for example Sphingolipids,sialo G _mL, or (B) and one or more hydrophilic polymers derive polyoxyethylene glycol (PEG) base for example.Be not subjected to any theoretical especially constraint, can think in this area, at least for containing Sphingolipids,sialo, sphingophospholipid, or the liposome of the spatial stability of PEG-deutero-lipid, the liposome enhanced circulating half-life of these spatial stabilities reduces (Allen etc. derived from picked-up in the cell of reticuloendothelium system (RES), FEBS Letters, 1987,223,42; Wu etc., Cancer Research, 1993,53,3765).

The liposome that comprises one or more various glycolipids known in the art.Papahadjopoulos etc. (Ann.N.Y.Acad, Sci., 1987,507,64) have reported Sphingolipids,sialo G _mL, gala E.C. 3.1.6.8 and phosphatidylinositols improve the ability of liposome blood halflife.These discoveries have been carried out detailed explanation by (Proc.Natl.Acad.Sci.U.S.A., 1988,85,6949) such as Gabizon.U.S. patent No. .4,837,028 and WO 88/04924, all be Allen etc., disclose and comprised (1) sphingophospholipid and (2) Sphingolipids,sialo G _mThe liposome of l or galactocerebroside sulfate ester.U.S. patent No. .5,543,152 (Webb etc.) disclose the liposome that comprises sphingophospholipid.Comprise 1, the liposome of 2-sn-two mnyristoyl Phosphorylcholines is open in WO 97/13499 (Lim etc.).

The liposome derived from the lipid of one or more hydrophilic polymers and preparation method thereof that much contains known in the art.Sunamoto etc. (Bull.Chem.Soc.Jpn., 1980,53,2778) illustrate the liposome that contains nonionic detergent, 2C _1215G, it contains the PEG base.Illum etc. (FEBS Lett., 1984,167,79) illustrate that the polyoxyethylene glycol hydrophilic coating of granules of polystyrene makes blood halflife improve greatly.The synthetic phospholipid of modification is by Sears (the U.S. patent No. 4,426,330 and 4,534,899) explanation by the carboxyl of additional polyalkylene glycol (for example PEG).Klibanov etc. (FEBS Lett., 1990,268,235) have illustrated that demonstration contains the experiment that can be significantly improved the blood circulation transformation period by PEG or PEG stearate deutero-phosphatidylethanolamine (PE) liposome.Blume etc. (Biochimica et Biophysica Acta, 1990,1029,91) expand above-mentioned observation to another kind of PEG-deutero-phosphatide, and DSPE-PEG for example is by being combined to form of DSPE (DSPE) and PEG.In European patent EP 0 445 131 B1 of Fisher and WO 90/04384, have illustrated at the covalently bound liposome of outside surface and PEG base.Contain 1-20mol% PE by PEG deutero-liposome composition and using method thereof in (U.S. patent No.s 5,013,556 and 5 such as Woodle, 356,633) and have illustrated among the Martin etc. (U.S. patent No. .5,213,804 and European patent EP 0 496 813 B1).The liposome that contains a large amount of other lipids-polymer scale zoarium has illustrated in WO 91/05545 and the U.S. patent No. 5,225,212 (all being Martin etc.) and WO94/20073 (Zalipsky etc.).The liposome that contains the ceramide lipid of PEG-modification has illustrated in WO 96/10391 (Choi et al).U.S. the patent No. 5,540, and 935 (Miyazaki etc.) and the U.S. patent No. 5,556,948 (Tagawa etc.) have illustrated the liposome that contains PEG, and it can further be derived with functional group on the surface.

Minority known in the art contains the liposome of nucleic acid.WO 96/40062 Thierry etc. discloses the method that is used for sealing at liposome high molecular nucleic acid.U.S. patent No. .5,264,221 Tagawa etc. disclose albumen-in conjunction with liposome and have asserted that the inclusion of this liposome can comprise dsRNA.U.S. patent No. .5,665,710 Rahman etc. have illustrated the method for sealing oligonucleotide in liposome.WO 97/04787Love etc. discloses that to comprise with the raf gene be the liposome of the dsRNAs of target.

Carrier (Transfersomes) is another kind of liposome, and is alterable height shape lipid coacervate, and it is the important candidate of drug delivery vehicle.Carrier can be illustrated as lipid liquid, and its alterable height shape makes it facilitate penetration of the hole littler than drop.Carrier adapts to its environment for use, for example its can self-optimizing (being suitable for the shape of skin mesopore), selfreparing, usually arrive its target and not division, and usually load (self-loading) automatically.Make the carrier can be with marginal surface-activator, normally tensio-active agent joins in the standard liposomal body composition.Carrier has been used to serum albumin is delivered to skin.Carrier-mediation sero-abluminous sent and demonstrated that to contain sero-abluminous solution the same effective with subcutaneous injection.

Tensio-active agent is found in formulation example such as emulsion (comprising micro emulsion) and the liposome and is widely used.Much the method for the most frequently used classification of tensio-active agent (natural or synthetic) not of the same race and character ordering has been utilized hydrophilic (HLB).The character of hydrophilic radical (being also referred to as ＂ ＂) is the most useful classification tool of using in the preparation of different surfaces promoting agent.(Rieger，in Pharmaceutical Dosage Forms，Marcel Dekker，Inc.，New York，N.Y.，1988，p.285).

If surfactant molecule can not ionization, it is divided into nonionogenic tenside.Nonionogenic tenside has extensive use in pharmacy and makeup, and can use in very wide pH value scope.Usually its HLB value scope be 2～about 18, depend on its structure.Ionic surfactant pack is drawn together the nonionic ester, glycol ester for example, propylene glycol ester, glyceryl ester, polyglycerol ester, sorboside ester, sucrose ester, and ethoxylated esters.Nonionic alkylamide and ether for example fat alcohol ethoxyl compound, the chimeric polymkeric substance of the pure and mild ethoxylated/propoxylated of propoxylation also in this class.Polyoxyethylene surfactant is the most frequently used nonionogenic tenside.

If surfactant molecule is electronegative when dissolving or being dispersed in the water, this tensio-active agent is divided into anion surfactant.Anion surfactant comprises for example soap of carboxylate salt; acyl-lactate, amino acid whose acid amides; sulfuric ester is the alkyl sodium sulfate salt of alkyl sodium sulfate salt and ethoxyquin for example, and sulfonate is Phenylsulfonic acid alkyl salt, acyl isethinate for example, acyl taurine salt and phosphoric acid salt.Most important anion surfactant is alkyl sodium sulfate salt and soap.

If surfactant molecule is positively charged when dissolving or being dispersed in the water, this tensio-active agent is divided into cats product.Cats product comprises quaternary ammonium salt and ethoxylated amine.The most frequently used in this class is quaternary ammonium salt.

If surfactant molecule can positively charged or negative electricity, this tensio-active agent is classified as amphoterics.Amphoterics comprises acrylic acid derivative, the alkylamide of replacement, N-alkyl betaine and phosphatide.

The purposes of tensio-active agent in medicament production, preparation and emulsion (Rieger, inPharmaceutical Dosage Forms, Marcel Dekker, Inc., New York, N.Y., 1988, commentary is arranged in p.285).

Penetration enhancers

In one embodiment, the present invention utilizes various penetration enhancers to realize nucleic acid (particularly dsRNAs) effectively is delivered on the skin of animal.Most medicines with the form of ionization or nonionicization exist with solution in.But, only have the lipid of being dissolved in or lipophilic medicine to pass cytolemma usually.Find if the film that passes was handled with penetration enhancers, even if be that non-oleophylic medicine also can pass cytolemma.Except helping the infiltration of non-oleophylic medicine to pass the cytolemma, penetration enhancers has also strengthened the perviousness of lipophilic medicine.

Penetration enhancers can be divided into a kind of in the 5 big classes, i.e. tensio-active agent, and lipid acid, biliary salts, sequestrant and non-chelating nonsurfactant (Lee etc., Critical Reviews in Therapeutic DrugCarriers Systems, 1991, p.92).Will describe above-mentioned each class penetration enhancers below in detail.

Tensio-active agent: related to the present invention, tensio-active agent (or ＂ surface-active agents ＂) is the chemical substance that reduces solution surface tension in being dissolved into the aqueous solution time or reduce the interfacial tension between the aqueous solution and other liquid, can strengthen the absorption of dsRNAs by mucous membrane.Except biliary salts and lipid acid, these penetration enhancers comprise, for example, lauric acid sodium sulfate, polyoxyethylene-9-lauryl and polyoxyethylene-20-cetyl ester) (Lee etc., Critical Reviews in Therapeutic Drug Carriers Systems, 1991, p.92); With perfluor chemistry emulsion, for example FC-43 (Takahashi etc., J.Pharm.Pharmacol., 1988,40,252).

Lipid acid: various lipid acid and derivative thereof as penetration enhancers comprise, for example, and oleic acid, lauric acid; capric acid (n-capric acid), tetradecanoic acid, palmitinic acid, stearic acid; linolic acid, linolenic acid, dicaprate, three decylates; XU 61518.10 (the single oleoyl of 1--rac-glycerine), GLYCERYL DILAURATE, sad; arachidonic acid, the single caprin of 1-, 1-dodecyl-aza-cycloheptane-2-ketone; fatty acyl carnitine, acyl group choline, its C ₁-C ₁₀Alkyl ester (for example, methyl esters, isopropyl ester and t-butyl ester), with its list-glyceryl ester and two-glyceryl ester (for example, oleic acid ester, laurate, decylate, myristinate, cetylate, stearate, linoleate, Deng) (Lee etc., Critical Reviews in Therapeutic Drug Carryier Systems, 1991, p.92; Muranishi, Critical Reviews in Therapeutic Drug Carriers Systems, 1990,7,1-33; El Hariri etc., J.Pharm.Pharmacol., 1992,44,651-654).

Biliary salts: biliary physiological role comprises that promotion disperses and absorption lipid and liposoluble VITAMIN (Brunton, Chapter 38 in:Goodman ﹠amp; Gilman ＇ s The Pharmacological Basis ofTherapeutics, 9th Ed., Eds. such as Hardman, McGraw-Hill, New York, 1996, pp.934-935).Various natural biliary saltss and synthesis of derivatives thereof are as penetration enhancers.Therefore, term ＂ biliary salts ＂ comprises biliary any natural constituents and any synthesis of derivatives thereof.Biliary salts of the present invention comprises, for example, cholic acid (or the acceptable sodium salt of its pharmacy, Sodium cholic acid), Felacrinos (Suprachol), Septochol (Sodium desoxycholate), paddy ammonia cholic acid (paddy ammonia Sodium cholic acid), glycocholic acid (NaGC), sugared Septochol (sugared Sodium desoxycholate), taurocholate (Taurocholic acid sodium salt), tauroursodeoxycholic acid (sodium taurodeoxycholate), gallodesoxycholic acid (gallodesoxycholic acid sodium), Ursodeoxycholic Acid (UDCA) (UDCA), ox sulphur-24,25-dihydro-Sodium Fusidate (STDHF), sugared dihydro Sodium Fusidate and polyoxyethylene-9-lauryl (POE) (Lee etc., CriticalReviews in Therapeutic Drug Carriers Systems, 1991, page 92; Swinyard, Chapter39 In:Remington ' s Pharmaceutical Sciences, 18th Ed., Gennaro, ed., MackPublishing Co., Easton, Pa., 1990, pages 782-783; Muranishi, Critical Reviews inTherapeutic Drug Carriers Systems, 1990,7,1-33; Yamamoto etc., J.Pharm.Exp.Ther., 1992,263,25; Yamashita etc., J.Pharm.Sci., 1990,79,579-583).

Sequestrant: sequestrant related to the present invention can be defined as by remove the compound of metal ion from solution with its formation complex compound, can strengthen the absorption of dsRNAs by mucous membrane.Consider it in the present invention as the purposes of penetration enhancers, the additional advantage of sequestrant is as the DNase inhibitor, because most DNA nuclease that characterizes needs divalent metal catalysis, thereby the agent that can be chelated suppresses (Jarrett, J.Chromatogr., 1993,618,315-339).Sequestrant of the present invention includes but not limited to disodium ethylene diamine tetraacetate (EDTA), citric acid, salicylate (for example, sodium salicylate, 5-methoxyl group sodium salicylate and homovanillic acid sodium (homovanilate)), the N-acyl derivative of collagen, N-aminoacyl derivative (the enamine) (Lee etc. of alkyl polyoxyethylene ether-9 and beta-diketon, Critical Reviews in TherapeuticDrug Carriers Systems, 1991, page 92; Muranishi, Critical Reviews in TherapeuticDrug Carriers Systems, 1990,7,1-33; Buur etc., J.Control Rel., 1990,14,43-51).

Non-chelating nonsurfactant: used here non-chelating nonsurfactant infiltration enhancing compound can be defined as the remarkable activity that does not show sequestrant or tensio-active agent but strengthen the compound (Muranishi of dsRNA by the absorption of digestion mucous membrane, Critical Reviews in Therapeutic DrugCarriers Systems, 1990,7,1-33).This class penetration enhancers comprises, for example, and unsaturated cyclohexyl urea, 1-alkyl-and 1-alkynyl nitrogen heterocyclic-aliphatic ketone derivative (Lee etc., Critical Reviews inTherapeutic Drug Carriers Systems, 1991, page 92); With the antiphlogistic of on-steroidal, diclofenac sodium for example, indomethacin (indomethacin) and phenylbutazone (Yamashita etc., J.Pharm.Pharmacol., 1987,39,621-626).

The reagent that strengthens the dsRNAs picked-up in the cell rank can also join in pharmacy of the present invention and other compositions.For example, known cation lipid, for example liposome (Junichi et al, U.S. patent No. .5,705,188), positively charged ion glycerol derivative and polycation molecule, polylysine (Lollo etc., PCT applies for WO 97/30731) for example also can strengthen the cellular uptake of dsRNAs.

Other reagent can be used to strengthen the infiltration of the nucleic acid of using, and comprise glycol, for example ethylene glycol and propylene glycol, and the pyrroles is 2-pyrroles for example, and azone and terpenes be limonene and piperitone for example.

Carrier

Certain composition of the present invention also comprises carrier compound in preparation.Here used ＂ carrier compound ＂ or ＂ carrier ＂ can refer to nucleic acid or its analogue, it is inertia (being not biologically active itself), but be identified as nucleic acid in the process in vivo, the nucleic acid by for example degradation bioactive or promote it from circulation, to get rid of the bioavailability of the nucleic acid that reduces biologically active.Nucleic acid and carrier compound are co-administered generally to be that the latter is excessive, can significantly reduce the amount of the nucleic acid that reclaims in liver, kidney or other outer circulation organs (extracirculatory reservoirs), the chances are, and general acceptor causes because carrier compound and nucleic acid are competed.For example, with polyinosinic acid, asuro, poly-cytidylic acid(CMP) (polycytidic acid) or 4-acetamido-4 ＇ isothiocyano-stilbene-2,2 ＇-when disulfonic acid was co-administered, thiophosphatephosphorothioate dsRNA can partly reduce (Miyao etc. in the recovery of hepatic tissue, Antisense Res.Dev., 1995,5,115-121; Takakura etc., Antisense ﹠amp; Nucl.Acid Drug Dev., 1996,6,177-183.).

Vehicle

Opposite with carrier compound, ＂ pharmaceutical carriers ＂ or ＂ vehicle ＂ are used for pharmacy acceptable solvent, suspension agent or any pharmacology inert carrier of one or more delivery of nucleic acids to animal.Vehicle can be a liquid or solid, and the form of using according to plan when mixing with other components of nucleic acid and institute's drug administration composition selects, and makes it become the volume that needs and denseness etc.General pharmaceutical carriers include but not limited to tackiness agent (for example, the W-Gum of pre-gelatinize, Polyvinylpyrolidone (PVP) or HYDROXY PROPYL METHYLCELLULOSE, etc.); Filler (for example lactose or other sugar, Microcrystalline Cellulose, pectin, gelatin, calcium sulfate, ethyl cellulose, polyacrylic ester or secondary calcium phosphate etc.); Lubricant (for example Magnesium Stearate, talcum, quartz, colloidal silicon dioxide, stearic acid, Metallic stearates, hydrogenated vegetable oil, W-Gum, polyoxyethylene glycol, Sodium Benzoate, sodium acetate etc.); Disintegrant (for example starch, sodium starch glycollate etc.); And wetting agent (for example Sodium Lauryl Sulphate BP/USP etc.).

There is not being suitable for the acceptable organic or inorganic vehicle of pharmacy that non-parenteral uses and being used to prepare composition of the present invention of untoward reaction with nucleic acid.Suitable pharmaceutically acceptable carrier includes but not limited to water, salts solution, alcohol, polyoxyethylene glycol, gelatin, lactose, amylose starch, Magnesium Stearate, talcum, silicic acid, sticking paraffin, Walocel MT 20.000PV, Polyvinylpyrolidone (PVP) etc.

The external preparation of nucleic acid can comprise for example alcohol of disinfectant and non-aqueous antiseptic solution, the non-aqueous solution in common solvent, or the solution of nucleic acid in the liquid or solid oil phase.Solution can comprise damping fluid, thinner and other suitable additives.Also can use the acceptable organic or inorganic vehicle of pharmacy that non-parenteral is used that is suitable for that does not have untoward reaction with nucleic acid.

The acceptable vehicle of suitable pharmacy includes but not limited to water, salts solution, alcohol, polyoxyethylene glycol, gelatin, lactose, amylose starch, Magnesium Stearate, talcum, silicic acid, sticking paraffin, Walocel MT 20.000PV, Polyvinylpyrolidone (PVP) etc.

Be fit to be delivered to the pharmaceutical composition of respiratory tract

The present invention provides on the other hand IRNA reagent will be delivered to respiratory tract, especially for the treatment cystic fibrosis.Respiratory tract comprises that the upper respiratory tract (comprising oropharynx and larynx) is that (comprise tracheae, bifurcated enters segmental bronchus and bronchiole to lower respiratory tract then then.Last lower respiratory tract is called conduction gas circuit (conductiveairways).The bronchiole terminal is divided into alveolar bronchiole (respiratory bronchioli), arrives last respiratory region, alveolar or lung inside (deep lung) then.Lung inside or alveolar are that the inhaling type that is used for systemic delivery iRNA reagent is treated aerocolloidal main target spot.

The pulmonary delivery composition can suck dispersion by the patient and send, and makes composition in the dispersion, and preferred iRNA reagent can reach lung, can directly enter circulation of blood by alveolar in lung.Pulmonary delivery is all effective for systemic delivery and local delivery treatments pulmonary disorder.

Pulmonary delivery can realize by different methods, comprises and uses micelle atomization, aerosolized and dry powder based formulation; It can be per os and/or intranasal that suction is used.Send and to realize with liquid spray, aerosol-Ji inhalation and dry powder diverting device.Preferred dose measuring apparatus (Metered-dosedevices).One of benefit of using atomizer or sucker is because device is self-contained, so can be with minimize contamination.The dry powder diverting device is sent the medicine that is mixed with dry powder easily.The iRNA composition can with from as lyophilized powder or spraying dry powder or with stable preservation of form of suitable powder carrier combination.Sending of composition for inhalation can be by quantitative time element mediation, described element comprises timer, dosimeter, time measurement device or time marker, and it is added in the device can follow the trail of dosage, monitoring adaptability and/or dosage triggering (dose triggering) to the patient during aerosol drugs is used.

The example that is used for the pharmaceutical devices of aerosol delivery comprises dose measurement sucker (MDIs), Diskus (DPIs) and air-blast atomizer.The exemplary suction delivery system that can be used to send iRNA reagent of the present invention is for example, U.S. Patent number 5,756,353; 5,858,784; With PCT application WO98/31346; WO98/10796; WO00/27359; WO01/54664; Explanation is arranged among the WO02/060412.Can be used to send other aerosol preparations of iRNA reagent in U.S. Patent number 6,294,153; 6,344,194; 6,071,497 and PCT application WO02/066078; WO02/053190; WO01/60420; Explanation is arranged among the WO00/66206.And then being used to send the iRNA compositions and methods can be according to being used in the method (for example antisense oligonucleotide) of sending other oligonucleotide that sucks, for example at Templin etc., and AntisenseNucleic Acid Drug Dev, 2000,10:359-68; Sandrasagra etc., Expert Opin Biol Ther, 2001,1:979-83; Sandrasagra etc., Antisense Nucleic Acid Drug Dev, 2002, illustrate among the 12:177-81.

Sending of reagent of the present invention can also comprise using of so-called ＂ prodrug ＂, promptly, the preparation of therapeutant or chemically modified, this therapeutant need the processing of certain form or need the transportation of the born system of target organism and could discharge the preferably action site release of expecting; The embodiment of back can be sent collaborative the use with respiratory tract, also can use with other embodiments of the present invention.For example, Ren Lei lung can be at the several minutes hydrolysis splitted deposition gas colloidal sol of removing or degrade fast within several hours.In the upper respiratory tract, the ciliate epithelial cell helps ＂ mucomembranous cilium motion (mucociliary excalator) ＂, and by such motion, particle is inswept in the mouth from respiratory tract.Pavia, D., ＂ Lung MucociliaryClearance, " in Aerosols and the Lung:Clinical and Experimental Aspects, Clarke, S.W. and Pavia, D., Eds., Butterworths, London, 1984.In lung inside, pulmonary alveolar macrophage can just be engulfed particle after particle deposition.Microscopy Res.Tech. such as Warheit, 26:412-422 (1993); And Brain, J.D., ＂ Physiology and Pathophysiology of PulmonaryMacrophages, ＂ in The Reticuloendothelial, S.M.Reichard and J.Filkins, Eds., Plenum, New.York., pp.315-327,1985.

In preferred embodiment, when particularly needing iRNA reagent systemic administration, aerosolized iRNA reagent is made microparticle.The microparticle of diameter 0.5～10 μ m can permeate lung, passes most natural cover for defenses.Requiring diameter is in order to walk around larynx less than 10 μ m; Require diameter be 0.5 μ m or above be for fear of exhalation.

Other components

Composition of the present invention also contains other adjuvant components of the routine of the consumption level that this area is determined in the pharmaceutical composition.Therefore, for example, composition can contain extra, compatible, pharmaceutically active substances, for example, pruritus, astringent matter, local anesthetic or antiphlogistic drug, maybe can contain and be useful on other materials that physics is prepared the various formulations of composition of the present invention, for example dyestuff, seasonings, sanitas, antioxidant, opalizer, thickening material and stablizer.But, should be unable to too influence the biological activity of composition components of the present invention adding fashionable, such material.Preparation can be a disinfectant, and if desired, can mix with auxiliary agent, for example lubricant, sanitas, stablizer, wetting agent, emulsifying agent, salt, thus influence osmotic pressure, buffering, color, taste and/or bad interactional aromatoising substance etc. does not take place with the nucleic acid of preparation.

Aqeous suspension can contain the material that increases suspension viscosity, for example comprises Xylo-Mucine, sorbyl alcohol and/or dextran.Suspension can also contain stablizer.

Some embodiment of the present invention provides a kind of pharmaceutical composition, and it contains (a) one or more dsRNA reagent and (b) one or more other chemotherapeutics, and it is by the effect of non-RNA interference mechanism.The example of such chemotherapeutics includes but not limited to daunorubicin, daunomycin, gengshengmeisu, Zorubicin, epirubicin, idarubicin, 4 ' deoxidation Zorubicin (esorubicin), bleomycin, Mafosfamide, ifosfamide, cytosine arabinoside, two-the chloroethyl nitrourea, busulfan, ametycin, radiating streptozotocin D, mithramycin, mithramycin, hydroxyprogesterone, testosterone, tamoxifen, dacarbazine, procarbazine, altretamine, pentamethyl-melamine, mitoxantrone, amsacrine, Chlorambucil, the methyl cyclohexane nitrourea, mustargen, melphalan, endoxan, Ismipur, 6-thioguanine, cytosine arabinoside, U-18496, hydroxyurea, Deoxycofomycin, 4-hydroxyl peroxide endoxan, 5 FU 5 fluorouracil (5-FU), floxuridine (5-FUdR), methotrexate (MTX), colchicine, taxol, vincristine(VCR), vinealeucoblastine(VLB), etoposide (VP-16), trimetrexate, irinotecan, topotecan, gemcitabine, Vumon, cis-platinum and diethyl diethylstilbestrol (DES).Usually, see The Merck Manual of Diagnosis and Therapy, 15th Ed.1987, pp.1206-1228, Berkow etc., eds., Rahway, N.J.When using with compound of the present invention, such chemotherapeutics (for example can use separately, 5-FU and oligonucleotide), use in order (for example, 5-FU and oligonucleotide for some time, be MTX and oligonucleotide then), or be used in combination (for example, 5-FU, MTX and oligonucleotide with one or more other such chemotherapeutics, or 5-FU, radiotherapy medicine and oligonucleotide).Antiphlogistic drug (including but not limited to non-steroidal anti-inflammatory medicine and corticosteroid) and antiviral drug (including but not limited to ribavirin, vidarabine, acyclovir and ganciclovir) also can be combined in the composition of the present invention.Usually see The Merck Manual of Diagnosis and Therapy, 15th Ed., Berkow etc., eds., 1987, Rahway, N.J. is respectively at 2499-2506 page or leaf and 46-49 page or leaf).Other non-dsRNA chemotherapeutics also within the scope of the invention.The compound of two kinds or above combination can use or use in order together.

The toxicity of such compound and result of treatment can be definite in microbial culture or laboratory animal by the pharmaceutical procedures of standard, for example, and in order to determine LD50 (50% lethal dosage in the colony) and ED50 (dosage of effective treatment 50% in the colony).Ratio between toxicity and the result of treatment is the treatment coefficient, and can be expressed as the ratio of LD50/ED50.The compound that the preferred therapeutic coefficient is high.

The data that cell culture test and zooscopy obtain can be used to prepare human dosage.The dosage of the present composition is generally at ED50 and only have in a small amount or do not have in the toxic circulation composition scope.Can change dosage according to used formulation and used route of administration.For used in the methods of the invention compound, the treatment significant quantity can be by cell culture test according to a preliminary estimate.Can in animal model, prepare dosage, obtain the circulating plasma concentration range of compound, perhaps in suitable, obtain target sequence polypeptide product the circulating plasma concentration range (for example, the peptide concentration that realize to reduce), be included in and measure IC50 (being the concentration of test compounds when reaching half maximum inhibition to symptom) in the cell cultures.Such information can be used for more accurately determining the effective dosage of the mankind.Can be by for example level of high-efficient liquid phase color spectrometry in blood plasma.

Except above-mentioned discussion use separately or composite application, dsRNAs of the present invention can use with the agent combination of other known effective treatment influenzas.Under any circumstance, the doctor can be based on utilizing the observed result of canonical measure usefulness known in the art to adjust amount and the time that dsRNA uses.

Methods of treatment and route of delivery

The composition local delivery that can will comprise iRNA reagent (for example targeting is in the iRNA of influenza virus reagent) by number of ways to action site or systemic delivery to being tried in the body.Exemplary approach comprises and directly locally applies to the affected part that for example lung and nasal passage and vein, nasal cavity, oral cavity and eyes are sent.The optimal way that is used to use iRNA reagent of the present invention is by being applied directly to the mode of lung and nasal passage with liquid, aerosol or spray solution.

Usually, sending of iRNA reagent of the present invention is iRNA reagent to be finished be delivered to the process of being tried intravital infection site.Finish this optimal way of sending and can for example be delivered to respiratory tissue, or carry out systemic administration by for example parenteral administration by topical application to lung or nasal passage by suction, spraying or intranasal administration.

Be used to suck or the formulation of parenteral administration is known for a person skilled in the art.Described formulation can comprise aseptic aqueous solution, also can contain damping fluid, diluent and other suitable additive in this solution.Use for carrying out intravenously, the total concn of reply solute is controlled so that it possesses the isotonicity of preparation.

Active compound disclosed herein preferably is applied to lung or the nasal passage that is tried body by the mode of any suitable.But can finish using by the aerosol suspensoid of using the respiratory particle (it is sucked by trying body) that contains one or more active compounds to active compound.Active compound can by aerosolized be various ways, for example: inhalation of Foradil Aerolizer formoterol fumarate, predetermined dose (metered dose inhalant) or liquid/liquid suspension, but be not limited thereto.But the respiratory particle can be liquid or solid.As the U.S. patent No. 4,501, described in 729, described particle can with selected compound optional comprise can effectively suppress from the passage mucus secretion, to absorb again other therapeutic component of the amount of water such as guanamprazine, phenmethyl amiloride or phenyl amiloride etc.

Granular pharmaceutical composition can be chosen wantonly with carrier and combine to assist dispersion or transhipment.Can will mix mutually with one or more active compounds such as (for example glucose, lactose, sucrose, trehalose, N.F,USP MANNITOL) suitable carriers such as sugar with the ratio (for example weight ratio of 1:1) of any suitable.

Contain and be useful on the particle of implementing activeconstituents of the present invention and should comprise the particle that to breathe size, also, can suck the alveolar that also can enter segmental bronchus and lung by mouth or nose and larynx thereby the particulate size is enough little.Usually preferred particulates is of a size of about 1 to 10 micron (more particularly, size is less than 5 microns).The particle with the size that can not breathe that is contained in the aerosol is easy to be deposited on throat and is swallowed down, therefore preferably makes can not respiratory particulate quantity minimize in the aerosol.Use for nose, be present in the nasal cavity, preferably make particle size be in the scope of 10-500 micron for guaranteeing it.

The liquid medicine composition that is used to produce aerocolloidal active compound can be by preparing active compound with making up such as suitable vehicle such as aseptic no heat source water.Be used to implement hypertonic saline solution of the present invention and be preferably aseptic apyrogenic solution, contain the acceptable salt of 1～15 weight % physiology, and preferably contain the acceptable salt of 3～7 weight % physiology.

The aerosol that can prepare the liquid particle that contains active compound, but the jet-type atomizer or the ultrasonic sprayer that drive of applying pressure for example by the mode of any suitable.This content for example can be referring to the U.S. patent No. 4,501,729.Atomizer is the device of commercially available acquisition, and its form by pressurized gas (being generally air or oxygen) being quickened pass aperture or the form by ultrasonic agitation are converted into the pharmacy aerosol spray with the solution or the suspension of activeconstituents.

The suitable form formula that is used for atomizer is made up of liquid carrier and activeconstituents, and the content of activeconstituents is up to 40% w/w of form formula, but preferably is lower than 20% w/w.Carrier is generally the water-alcoholic solutions of water (most preferably being aseptic no heat source water) or dilution, is preferably to wait to ooze carrier, but can sodium-chlor makes it become the carrier that relative body fluid oozes for height by for example adding.Optional additive comprises sanitas such as methyl hydroxybenzoate (use when described form formula is not aseptic makings), antioxidant, correctives, volatile oil, buffer reagent and tensio-active agent.

The aerosol that contains the solid particulate of activeconstituents can adopt arbitrarily solid particulate therapeutical agent aerosol dispenser to make similarly.But be used for to being tried the aerosol that aerosol dispenser that body uses the solid particulate therapeutical agent produces the particle of respiratory with the speed that is suitable for the people and uses and generates the certain volume of the therapeutical agent that contains predetermined dose.An exemplary type of solid particulate aerosol dispenser is an insufflator.Be used for comprising pulverizing powder by the suitable form formula that insufflation is used, it can be sent by insufflator, maybe can be absorbed into nasal cavity by the mode of snuff.In insufflator, make powder (for example, effectively carry out the powder of the predetermined dose of treatment described herein) be included in usually by in the made capsule or capsule (cartridge) of gelatin or plastics, described capsule or capsule are pierced in position or open, suck so that powder is attracted by air and passes through device, and send, perhaps send by mechanical pump.The powder of packing in the medicine absorber can be made up of activeconstituents separately, perhaps by containing activeconstituents, forming such as the powdered mixture of suitable powder diluent such as lactose and optional tensio-active agent.Usually contain 0.1 to 100w/w activeconstituents in the form formula.

Second type exemplary aerosol dispenser comprises the sucker of predetermined dose.The sucker of predetermined dose is the aerosol dispenser of pressurization, contains the suspension or the solution dosage of activeconstituents usually in the liquefaction propeller.Use these whens device, can by valve come delivery formulations so that preparation send to set volume (being generally 10) to 200ul thus preparation contains the particle spraying of described activeconstituents.Suitable thruster comprises for example some chlorofluorocarbon compound of Refrigerant 12, trichlorofluoromethane, dichloro tetrafluoro ethane and composition thereof etc.Described preparation also can further contain one or more cosolvent (for example ethanol), tensio-active agent (for example oleic acid or sorbitan trioleate), antioxidant and suitable correctives.

IRNA reagent can join in the pharmaceutical composition that is suitable for using.For example, composition can comprise one or more iRNA reagent and pharmaceutically acceptable carrier.Here used " pharmaceutically acceptable carrier " refers to comprise any and all and pharmacy are used compatible solvent, dispersion medium, coating, antiseptic-germicide and anti-mycotic agent, isobaric absorption delay agent etc.The purposes of the reagent of known such medium of those skilled in the art and pharmaceutically active substances.Except with inconsistent any conventional media of active compound or reagent, can expect its purposes in composition.The active compound that replenishes also can join in the composition.

Using can be by being tried body or waiting other people to carry out such as the care-giver.The care-giver can be any main body that the people is provided nurse or looks after.For example, hospital, collecting post, Doctor's office, outpatient's ambulatorium; Such as doctor, nurse or other medical workers such as practitioner; Or be guardians such as spouse or father and mother.Can provide pharmacological agent by quantitative dosage, or adopt the divider of sending predetermined dose that pharmacological agent is provided.

Term " treatment significant quantity " refers to the existing following amount of composition: thus this amount obtains the physiological responses of expection in order to the composition of medicine that desired level is provided in the body in to be treated trying.

Term " physiology significant quantity " thus referring to be delivered to is tried to produce in the body the required alleviation or the amount of result of treatment.

Term " pharmaceutical acceptable carrier " refers to described carrier and can be absorbed in the lung and can not cause significant toxic side effect to lung.

Term " is used altogether " and is referred to described two or more reagent, especially two or more iRNA agent administration to being tried in the body.Described reagent can be included in the single pharmaceutical composition and can use simultaneously, and perhaps described reagent can be included in the isolating form formula and can be applied in proper order and be tried in the body.As long as two kinds of reagent can be detected simultaneously, just we can say that these two kinds of reagent use simultaneously in being tried the body body.

The kind that can be used as the drug excipient of carrier comprises: such as human serum albumin stablizers such as (HSA); Such as weighting agents such as carbohydrate, amino acid and polypeptide; PH adjusts agent or buffer reagent; Such as salts such as sodium-chlor; Or the like.These carriers can or be amorphous for crystal type, or are this mixture of two kinds.

Valuable especially weighting agent comprises the combination of carbohydrate, polypeptide, amino acid or the above-mentioned substance of consistency.Suitable carbohydrate comprises: monose such as semi-lactosi, D-seminose, sorbose; Disaccharides such as lactose, trehalose; Cyclodextrin such as 2-hydroxypropyl-beta-cyclodextrin; Polysaccharide such as raffinose, maltodextrin, dextran; Aldehyde alcohol such as N.F,USP MANNITOL, Xylitol; Or the like.Comprise lactose, trehalose, raffinose, maltodextrin and N.F,USP MANNITOL in the preferred group of carbohydrate.Suitable polypeptide comprises L-aspartyl-L-phenylalanine methylester.Amino acid comprises L-Ala and glycine, preferred glycine.

Suitable pH regulator agent or buffer reagent comprise that for example Trisodium Citrate, sodium ascorbate etc. utilize the organic salt of organic bronsted lowry acids and bases bronsted lowry preparation, optimization citric acid sodium.

Dosage

IRNA reagent can be used less than the unitary dose of about 75mg/ kg body weight, or with less than about 70,60,50,40,30,20,10,5,2,1,0.5,0.1,0.05,0.01,0.005,0.001 or the dosage of 0.0005mg/ kg body weight use, and with iRNA reagent (for example about 4.4 x 10 less than 200nmol ¹⁶Copy (copies))/dosage of kg body weight uses or with less than 1500,750 300,150,75,15,7.5,1.5,0.75,0.15,0.075,0.015,0.0075,0.0015,0.00075, the dosage of iRNA reagent/kg body weight of 0.00015nmol is used.Described unitary dose for example can be used by injection (for example in vein or intramuscular, the sheath or directly to organ), suction or external application.

IRNA reagent is about 0.00001mg～about 3mg/ organ to the dosage that organ (for example, to lung) is used directly, or preferably about 0.0001-0.001mg/ organ, about 0.03-3.0mg/ organ, about 0.1-3.0mg/ eye or about 0.3-3.0mg/ organ.

Described dosage can be the amount that disease or disturbance state are effectively treated or prevented.It can be used as prevention or provide as main or part prescription.

In one embodiment, unitary dose can be less than once a day, for example is less than per 2,4,8 or 30 days once.In other embodiment, unitary dose is also used not according to frequency (for example, regularly frequency).For example, unitary dose can single administration.Because can using the back at the iRNA reagent composition, the silence of iRNA reagent mediation continues several days time, thereby in many cases, can come applying said compositions with the frequency that is less than once a day, perhaps, in some cases, can in whole treatment plan, only use once.

In one embodiment, used the iRNA reagent of initial dose and one or many maintenance dose to trying body, described iRNA reagent for example is double-stranded iRNA reagent or siRNA reagent (for example, such as precursor such as the bigger iRNA reagent that can be processed into siRNA reagent or for encoding such as the DNA of the iRNA reagent of double-stranded iRNA reagent or siRNA reagent or its precursor etc.).Maintenance dose (one or more) is usually less than initial dose, for example is below half of initial dose.The mode of keeping can comprise with one or more dosage in the dosage range in 0.01～75mg/ kg body weight/sky to be handled trying body, and this dosage for example can be 70,60,50,40,30,20,10,5,2,1,0.5,0.1,0.05,0.01,0.005,0.001 or 0.0005mg/ kg body weight/sky.Maintenance dose is preferably per 5,10 not to be higher than, or 30 days frequencies are once used.Further, this treatment plan can continue for some time, and this time is depended on specified disease characteristic, severity of disease and patient's overall condition.In preferred embodiment, described dosage can be not higher than once a day (for example be not higher than per 24,36,48 or more hours once, as not being higher than per 5 or 8 days once) frequency send.After treatment, can monitor and the alleviation of disease symptoms is monitored the variation of its situation.When the patient does not have significant reaction to current dosage level, can increase the dosage of described compound; Perhaps when the alleviation of observing disease symptoms or morbid state have disappeared or observed the side effect of not wishing to occur, should reduce described dosage.

Significant quantity can be as required or suitable consideration under particular case, takes separately or use with two kinds or above dosage.Promote to repeat if desired or frequently inculcate, implant delivery apparatus, for example, (for example can use pump, semipermanent support (stent), intravenously, intraperitoneal, (intracisternal) or capsule interior (intracapsular) in the brain pond), or bank (reservoir).

After successfully treating, need keep treatment to the patient, to prevent the illness recurrence, compound wherein of the present invention is used (seeing US 6,107,094) with the maintenance dose of 0.001～100g/ kg body weight.

The concentration of iRNA reagent composition is to treatment or human illness or the enough effective amount of adjusting physiological situation of prevention.The concentration of the iRNA reagent of being used or amount depend on the determined parameter of reagent and application process (for example nose is used, oral administration or lung use).For example, thus nose uses that the concentration that trends towards making some compositions is lower avoids making nasal passage to be upset or to burn.Sometimes need oral dosage form is diluted 10～100 times so that suitable asal agent type to be provided.

Some factor may influence being tried the dosage that body is effectively treated, and described factor includes but not limited to: the severity of disease or illness, previous treatment, the general health situation of being tried body and/or age and other disease of being had etc.If the effective dose such as iRNA reagent such as siRNA reagent that is used for the treatment of can increase along with the process of special treatment or reduce to some extent, then also is preferred.The variation of dosage may be because due to the result of diagnostic assay, and is obviously interpretable by the result of diagnostic assay.For example, after using the iRNA reagent composition, can monitor being tried body.According to the information that monitoring obtains, can use the iRNA reagent composition of additional content.

Dosage depends on the seriousness and the reactivity of illness to be treated, through in a few days to the treatment of some months, or up to curing or reaching mitigate the disease.Can calculate optimal dose planning in the intravital accumulation of patient from measuring medicine.Those skilled in the art are easy to determine optimal dose, quantivative approach and repetition rate.Optimal dose can change according to the relative efficiency of each compound, and can estimate based on effective EC50s in the above-mentioned animal model in vitro and in vivo.

The present invention will be described hereinafter will to utilize embodiment, but described embodiment should not be construed as further restriction of the present invention.

Embodiment

Below represent nucleotide sequence with abbreviation concrete in standardized denomination and the table 3.

The abbreviation of the nucleotide monomer that table 3 uses in nucleotide sequence is represented.Should be appreciated that when these monomers are in oligonucleotide it is interconnective with 5 ＇-3 ＇-phosphodiester bond.

Abbreviation a	Nucleotide
		A，a	2 ＇-deoxidation-adenosine-5 '-phosphoric acid ester, adenosine-5 '-phosphoric acid ester
C，c	2 ＇-deoxidation-Cytidine-5 '-phosphoric acid ester, Cytidine-5 '-phosphoric acid ester
		G，g	2 ＇-deoxidation-guanosine-5 '-phosphoric acid ester, guanosine-5 '-phosphoric acid ester
T，t	2 ＇-deoxidation-thymidine-5 '-phosphoric acid ester, thymidine-5 '-phosphoric acid ester
		U，u	2 ＇-deoxidation-uridine-5 '-phosphoric acid ester, uridine-5 '-phosphoric acid ester
N，n	Any 2 ＇-deoxidation-Nucleotide/Nucleotide (G, A, C, or T, g, a, c or u)
		am	2 ＇-O-methyladenosine-5 '-phosphoric acid ester
cm	2 ＇-O-methylcytidine-5 '-phosphoric acid ester
		gm	2 ＇-O-methylguanosine-5 '-phosphoric acid ester
tm	2 ＇-O-methyl-thymidine-5 '-phosphoric acid ester
		um	2 ＇-O-methyluridine-5 '-phosphoric acid ester
A， C， G， T， U， a， c， g， t， u	Underscore: nucleosides-5 '-thiophosphatephosphorothioate
		x	Universal base

^aCapitalization is represented 2 ＇-deoxyribonucleotide (DNA), and lowercase is represented ribonucleotide (RNA)

The source of reagent

For the situation that the source of reagent does not provide specially, this reagent can obtain in the molecular biology reagent suppliers that quality/purity reach the molecular biology application standard from any one.

Embodiment 1: select sequence

Carry out the siRNA design to determine the proteic influenza A mRNAs of MP, NP, PA, PB1 and PB2 of siRNAs target.In the first round, the electronic selection of siRNA (insilico selection) has the sequence of the sequence of 44 target MP, 3 target NP and the sequence of 1 target PB1.Influenza A gene PA or PB2 there are not specific siRNAs, require 80% target coverage (target coverage) and 80% target efficient (target efficency) (consulting hereinafter) by the first step chosen process.

In order to build the environment of sequential analysis, from Ftp: //ftp.virginia.edu/pub/Last download fastA software package (Pearson, W.R.; Lipman, D.J., PNAS 1988, and 85:2444), the standard that is installed on is installed the Suse that is provided with

9.3 on the workstation under the operating system.In order to move perl script (perl scripts), guarantee to work at 9.3 times perl compilers of Suse Linux (perl interpreter) (version 5.8.6 (copyright 1987-2004, Larry Wall)) that standard is provided with.Brown Lab Web server (Brown Lab Web Server) from North Carolina State University website is downloaded BioEdit sequence alignment editor (Hall, T.A., Nucl.Acids.Symp.Ser.1999,41:95), and be installed on Microsoft Windows2000

On the computer under the operating system.

The workflow of electronic selection is as follows: the influenza A sequence that download to need, compare and produces statistic data to obtain base distribution with respect to the consensus sequence of calculating on each position.The candidate target area that utilizes the identification of perl script to satisfy the choice point standard (cut-off criteria) of definition.Utilize the specificity of the siRNA sequence of fastA Algorithm Analysis candidate target area to people RefSeq database.SiRNAs is marked according to the specificity of prediction with another perl script.At last, the siRNAs of specific criteria is satisfied in artificial selection.

The influenza A sequence that needs was downloaded from NCBI influenza virus database on June 24th, 2005, and download site is the website of National Center for Biotechnology Information.MP, NP, PB1, the number of each gene of PB2 and PA is shown in the table 4, and corresponding registration number provides in table 4.

The gene order number of used influenza gene M P, NP, PB1, PB2 and PA from various virus subtypes in the table 4:siRNA sequence electronic selection

Gene H1N1 H2N2 H3N2 H5N1 H7N3 H7N7 H9N2 sum

MP 10 2 13 166 28 16 128 363

NP 12 3 10 169 12 6 138 350

PB1 3 2 10 163 10 7 127 322

PB2 2 1 10 164 12 9 133 331

PA 2 2 10 171 11 8 124 328

Utilize the default parameters of each target respectively, with the ClustalW multiple ratio of BioEdit sequence alignment editor to function (Thompson, J.D. is etc., Nucleic Acids Res.1994; 22:4673), produce the overall comparison of all sequences.Obtain position Nucleotide numerical value summary output (positional nucleotidenumerical summary output), the information that provides on each position the base with respect to the consensus sequence of the calculating of each target to distribute.

Determine that length is that the choice point standard definition of candidate's target area of 19 Nucleotide is:

Standard 1, target covers: at least 80% the available all sequences of each influenza A gene need be embodied in the candidate region.

Standard 2, target efficient: at least 80% all sequences-wherein the candidate region is embodied, and need be identical in described candidate region.

Definition standard 1 does not have sequence information in the zone.Standard 2 is guaranteed a large amount of hypotypes of target.

With the Nucleotide numerical value summary file of perl script screening position, be the candidate target area of 19 bases and be created in the file that is used as the fastA input in subsequently the analytical procedure to determine the meeting cut-out standard and to have length.For the script input, import the sequence sum of each target, and the value of the conservative per-cent of input is 80.Select corresponding in candidate's target region all candidates of common sequence adopted siRNA arranged, and be stored in the file of fastA form.In order to consider the interaction between siRNAs potential dTdT overhang and the target sequence, all sequences are expanded with " AA " at 5 ' end, produce the list entries of 21 mer.For each candidate's target region, produce the file that another has region characteristic, described region characteristic is: the number of target coverage (sequence of existence), target efficient, mispairing sum, conserved sequence and the sequence number of existence.

For further selection, candidate siRNAs is sorted according to the potentiality (potentiality of missing the target, off-target potential) with host's (at this, without limits, people) gene interaction of prediction.Suppose that the siRNAs with the low potentiality of missing the target has more specificity in vivo.

In order to predict the siRNA specificity potentiality of missing the target, do following hypothesis:

1) position 2-9 of chain (from 5 ' to 3 ' counting) than remaining sequence (non-seed region) more help to miss the target potentiality (Haley, B., and Zamore, P.D., Nat Struct Mol Biol.2004,11:599).

2) all calculate the score of missing the target for hitting (hit) each time, based on the identity of siRNA sequence and the position of mispairing.

3) by suitable nucleotide modification being introduced sense strand (for example, all Nucleotide that contain pyrimidine base are the Nucleotide that 2 ＇-O-methyl is modified), sense strand is become RNA is disturbed non-activity; Therefore, only need to consider the potentiality of missing the target of antisense strand.

In order to determine the potential gene that misses the target,, carry out the homology search to add 3 ' terminal AA tail (in order to explain the TT overhang) corresponding to candidate's target region 21mer sequence at public obtainable people mRNA sequence.For this reason, with all 21mer list entries, at people RefSeq database (on 07 27th, 2005 from Ftp: //ftp.ncbi.nih.gov/refseq/Downloaded available edition), carry out fastA (version 3 .4) search.Carry out the fastA search with parameter-value-right-b 30-g 30 (parameters-values-pairs-b 30-g 30), calculate the homology of 21 mer full length sequences thus.Search Results is with potential the missing the target of tabular form show candidate siRNA sequence.

For the potential list that misses the target to gained carries out sorting, the fastA output file is analyzed, identify host gene with the highest value of missing the target.Extract the following characteristic of missing the target of each 21mer list entries, miss the target for each is potential and calculate the score of missing the target.

1.21mer the number of the identical Nucleotide of sequence (identity)

2. the number of mispairing in the seed region

For the hypothesis of considering 1 and 2, by the following calculating score of missing the target:

Identity-0.2 ^*The number of seed mispairing

According to the highest score of missing the target (rising) of all siRNAs, it is carried out sorting.The choice point that the score of missing the target 16.8 is selected as siRNA.Article 42, the specific siRNAs of influenza A stromatin (MP), the specific siRNAs of 3 influenza A nucleocapsid protein (NP) and the specific siRNA of 1 influenza A polysaccharase basic protein 1 (PB1) have this threshold value or are lower than the score of missing the target of this threshold value.

In view of the number of the candidate siRNAs that produces by above-mentioned system of selection lower, with choice point that is set to 70% standard 1 (target fraction of coverage) and the standard 2 (target efficient) of keeping 80%, checked once more to be the Nucleotide numerical value summary of position, repeated the score ordering of missing the target mentioned above subsequently.For amounting to 48 candidate siRNAs, additionally selected 2 specific siRNA of influenza A MP mRNA, its target efficient is 79.9%.These 48 candidate siRNAs sequences are shown among the table 1A.

Because system of selection mentioned above only produces the candidate agent of limited quantity, thus choice criteria relaxed a little, to produce other candidate agent.Specifically, standard 1 is above relaxed target coverage to 50%, and standard 2, target efficient remains on 80%, and repeats above-mentioned chosen process.This method has produced extra reagent A L-DP-8001 to AL-DP-8040, and it is listed among the table 1C.

In this process, can recognize, miss the target to such an extent that in potential reagent, cause maximum loss rate (attrition rate) step by step.In order to obtain more candidate agent, thereby with the standard 1 of 80% target coverage, the standard 2 of target efficient 80% repeats chosen process again one time, and ignores the score of missing the target.The extra preparation that this process obtains is listed among the table 1D.But list in other the candidate agent of table among the 1E and be by once more with the standard 2 of the standard 1 of 50% target coverage, 80% target efficient and ignore the score of missing the target and repeat this selection and obtain.

By allowing to introduce free base, determine other candidate iRNA reagent.When each chain in the non-seed region (corresponding to the position 2-9 of antisense strand) at iRNA reagent is introduced 3 free bases at most, determine at first that with the perl script target coverage and target efficient are 100% candidate sequence.Table 1F shows the reagent of determining in this way.In second takes turns, determine extra iRNA, described iRNA has 80% target coverage and target efficient when allowing to introduce 1 free base.These iRNA list among the table 1G.

Embodiment 2:siRNA is synthetic

Nucleotide synthetic that comprises natural base

Single stranded RNA s use Expedite 8909 synthesizers (Applied Biosystems, AppleraDeutschland GmbH, Darmstadt, Germany) with the scale solid phase synthesis of 1 μ mole, controllable bore diameter glass (CPG, 500

, Glen Research, Sterling VA) and as solid support.RNA and the RNA that contains 2 '-O-methyl nucleotide use respectively corresponding phosphoramidite and 2 '-O-methyl phosphoramidite by solid phase synthesis produce (Proligo Biochemie GmbH, Hamburg, Germany).These structural units (building blocks) are incorporated on the interior selected site of oligonucleotide chain-ordering scope, it utilizes standard nucleoside phosphoramidites chemistry, as at book " Current protocolsin nucleic acid chemistry ", Beaucage, S.L. etc. (Edrs.), John Wiley ﹠amp; Described in the Sons, Inc., New York, NY, USA.The thiophosphoric acid key is by being replaced by the iodine oxidizing agent solution Beaucage reagent in acetonitrile (1%) (Chruachem Ltd, Glasgow, UK) solution and introducing.Further auxiliary reagent available from Mallinckrodt Baker (Griesheim, Germany).

By the oligonucleotide crude product is carried out anionresin HPLC, carry out deprotection and purifying according to existing method.(Unterschlei β heim Germany), determines productive rate and concentration by each RNA solution in the UV absorption value at 260nm wavelength place for DU640B, Beckman Coulter GmbH to utilize spectrophotometer.Double-stranded RNA process and annealing buffer (20mM sodium phosphate, pH6.8; 100mM sodium-chlor) in etc. mole complementary strand solution mix mutually and form, 3-4 hour cool to room temperature passed through in and heating 3 minutes in 85-90 ℃ of water-bath then.Purified RNA solution is standby-20 ℃ of storages.

As the result of above-mentioned synthetic method, all do not have phosphate-based on 5 ＇-terminal nucleotide according to the above-mentioned oligonucleotide that is synthesized.

Nucleotide synthetic that comprises universal base

Synthesizing of the phosphoramidite of 5 '-O-(4,4 '-dimethoxytrityl)-2 '-O-(tert-butyl dimethylsilyl)-1 '-(5-nitroindoline base)-D-riboside and controlled hole glass carrier

Steps A: 1-O-methyl D-riboside (102).

Add the vitriol oil (1.88mL) to the solution of D-ribose (25g) in dry methyl alcohol (300mL), and at room temperature stirred 3 days.Then with reaction mixture with 1N sodium hydroxide solution neutralization and be condensed into thick residue.Thick residue is dissolved in the methyl alcohol (200mL), filters out solid.Filtrate is condensed into thick residue, is added to then on the silicagel column, obtain syrupy shape pure compound (23.0g, 82%) with methylene chloride-methanol (5:1) wash-out.

Step B:1-O-methyl-2,3,5-three-O-(2, the 4-dichloro benzyl)-D-riboside (103)

To 1-O-methyl D-riboside (13.43g, 81.83mmol), the solution of 18-hat-6 (1.34g) in dry THF (100mL) adds the potassium hydroxide powder (69g 1.23mol), and at room temperature stirs 40～60min.Drip 2, (51mL, stirred reaction mixture spends the night 4-dichloro benzyl chlorination thing 368.2mmol) and under uniform temp.Filter out solid, and filtrate is condensed into thick residue, be added to then on the silicagel column, obtain white solid pure compound (48g, 92%) with hexane-ethyl acetate (4:1) wash-out.

1H-NMR(CDCl ₃，400MHz)：δ 7.46-7.34(m，5H，ArH)，7.24-7.16(m，4H，ArH)，4.99(s，1H，H-1)，4.71(dd，2H，Jgem＝12.8Hz，OCH ₂Ar)，4.63-4.61(m，4H，2 OCH ₂Ar)，4.38-4.36(m，1H)，4.19-4.16(dd，1H)，3.98(d，1H，J＝4.4Hz)，3.75(dd，1H，J＝3.6，J＝10.2Hz，H-5a)，3.66(dd，1H，J＝3.6，J＝10.4Hz，H-5b)，3.37(s，3H，OCH ₃)。

Step B:1-bromo-2,3,5-three-O-(2, the 4-dichloro benzyl)-D-ribose (104).

To using ice bath refrigerative 1-O-methyl-2,3, (3.22g, 5.02mmol) cold soln in dry methylene chloride (50mL) adds HOAc-HBr (5.3mL, 30%) to 5-three-O-(2, the 4-dichloro benzyl)-D-riboside, and stirs 3h down at 0-25 ℃.Reaction mixture is condensed into thick residue, and obtains thick residue with toluene (3 x 30mL) coevaporation, dry and be used for next step reaction and need not purifying and evaluation under good vacuum condition, it is a syrupy shape.

Step D:1-(5-nitroindoline base)-2,3,5-three-O-(2, the 4-dichloro benzyl)-D-riboside (105).

(2.44g is 15.06mmol) at dry CH to the 5-nitroindoline ₃Solution among the CN (30mL) adds sodium hydride (602mg, 15.06mmol, 60%) and at room temperature stirs 3-4h in the argon gas.Be added in exsiccant CH ₃The saccharide donor (104) of the front gained among the CN (10mL), and under uniform temp, stir in the argon gas and spend the night.Filter out solid, and filtrate is condensed into thick residue, it is added in the silicagel column also obtains pure compound 105 (2.16g, 60%), as α and β mixture (1:1) with hexane-ethyl acetate (3:1) wash-out.

Step e, F:5 '-O-(4,4 '-dimethoxytrityl)-1 '-(5-nitroindoline base)-D-riboside (106) and (107)

To 1-(5-nitroindoline base)-2,3, (1.16g, 1.51mmol) cold soln in-78 ℃ dry methylene chloride (100mL) adds BCl to 5-three-O-(2, the 4-dichloro benzyl)-D-riboside 105 ₃At methylene dichloride (23mL, the 1.0M) solution in, and under uniform temp, stir 2h in the argon gas, and stir 2h at-40 ℃.Reaction mixture is with methyl alcohol-methylene dichloride (1:1,50mL) cooling and neutralize with ammonia-methanol solution.Filter out solid, and filtrate is condensed into thick residue, it is added in the silicagel column also obtains pure compound (300mg, 68%), as α and β mixture (1:1) with methylene chloride-methanol (10:1) wash-out.(840mg, 2.86mmol) solution in dry pyrimidine (3-4ml) and DMAP (90mg) adds DMTrCl (1.06g), and at room temperature stirs in the argon gas and spend the night to above-mentioned gained compound.Reaction mixture is condensed into thick residue, and it is added in the silicagel column, and obtains pure compound 106 (550mg) and compound 107 (190mg) with hexane-ethyl acetate (1:1) wash-out, compound 106 and 107 mixture (360mg).

Compound 106:1H-NMR (CDCl ₃, 2D g-COSY and 2D NOESY, 400MHz): δ 8.49 (d, 1H, J=1.6Hz), 8.35 (d, 1H), 8.03 (dd, 1H, J=2.0, J=9.0Hz), 7.70-7.69 (m, 2H), 7.47-7.14 (m, 8H, ArH), 6.86-6.81 (m, 5H, ArH), 6.71 (d, 1H, J=3.6Hz), 6.41 (d, J=5.2Hz, H '-1), (4.73 t, 1H, J=4.8Hz, H '-2), 4.46-4.42 (m, 3H, H '-3, H '-4, H '-5), 3.79 (s, 6H, 2OCH ₃), 3.51 (dd, 1H, J=3.2, J=10.4Hz, H '-5a), 3.26 (dd, 1H, J=3.2, J=10.6Hz, H '-5b.

Compound 107:1H-NMR (CDCl ₃, 2Dg-COSY and 2D NOESY, 400MHz): δ 8.55 (d, 1H, J=2.0Hz), 7.98 (dd, 1H, J=2.4, J=9.2Hz), 7.60 (d, 1H, J=9.2Hz), 7.53 (d, 1H, J=3.2Hz), 7.44-7.42 (m, 2H), and 7.34-7.24 (m, 7H, ArH), 6.84-6.81 (m, 4H, ArH), 6.68 (d, 1H, J=3.2Hz), 6.00 (d, 1H, J=5.2Hz, H '-1), 4.53 (t, 1H, J=7.6Hz), and 4.46-4.44 (m, 1H), 4.23-4.20 (m, 1H), 3.80-3.76 (m, 7H, 2OCH ₃, H '-5), 3.55 (dd, 1H, H '-5a), 3.43 (dd, 1H, H '-5b).

Step G:5 '-O-(4,4 '-dimethoxytrityl)-2 '-O-(tert-butyl dimethylsilyl)-1 '-(5-nitroindoline base)-D-riboside (108) and 5 '-O-(4,4 '-dimethoxytrityl)-3 '-O-(tert-butyl dimethylsilyl)-1 '-(5-nitroindoline base)-D-riboside (109)

To 5 '-O-(4,4 '-dimethoxytrityl)-1 '-(5-nitroindoline)-D-riboside (106) (550mg, 0.92mmol), AgNO ₃(188mg, 1.104mmol), and pyrimidine (0.74mL, 9.2mmol) solution in dry THF (9.2mL) adds TBDMSCl (188mg 1.196mmol), and at room temperature stirs in the argon gas and spends the night.Filter out solid, and filtrate is condensed into thick residue, it is added in the silicagel column, and obtain pure compound 108 (230mg with hexane-ethyl acetate (4:1) wash-out, 35%) mixture (110mg of compound 109 (150mg, 23%) and compound 108 and 109,, 17%), overall yield is 75%.

Compound 108:1H-NMR (CDCl ₃, 2D g-COSY, 2D NOESY, 400MHz): δ 8.56 (d, 1H, J=2.4Hz), 7.88 (dd, 1H, J=2.4, J=8.8Hz), 7.62 (d, 1H, J=9.2Hz), 7.54 (d, 1H, J=3.6Hz), 7.46-7.44 (m, 2H), 7.36-7.25 (m, 6H, ArH), 6.85-6.83 (d, 5H, ArH), 6.69 (d, 1H, J=3.6Hz), 5.94 (d, 1H, J=7.2Hz, H '-1), 4.69 (dd, 1H, H '-2), 4.31-4.29 (m, 2H, H '-3, H '-4), 3.80 (s, 6H, 2OCH ₃), 3.58 (dd, 1H, J=2.0, J=10.6Hz, H '-5a), 3.40 (dd, 1H, J=2.0, J=10.4Hz, H '-5b), 2.85 (d, 1H, J=0.8Hz, 3 '-OH), 0.78 (s, 9H, t-Bu) ,-.016 (s, 3H, SiCH ₃) ,-0.43 (s, 3H, SiCH ₃).

Compound 109:1H-NMR (CDCl ₃, 2D g-COSY, 2D NOESY, 400MHz): δ 8.61 (d, 1H, J=2.4Hz), 8.05 (dd, 1H, J=2.0, J=8.8Hz), 7.69-7.65 (m, 2H), 7.47-7.45 (m, 2H, ArH), 7.36-7.27 (m, 5H, ArH), 6.86-6.83 (m, 3H, ArH), 6.71 (d, 1H, J=3.2Hz), 5.99 (d, 1H, J=4.8Hz, H '-1), 4.51 (t, 1H, J=4.8Hz, J=5.6Hz, H '-3), 4.40-4.36 (m, 1H, H '-2), 4.17-4.15 (m, 2H, H '-4, H '-5), 3.82 (s, 3H, OCH ₃), 3.81 (s, 3H, OCH ₃), 3.63 (dd, 1H, J=2.4, J=11.0Hz, H '-5a), 3.31 (dd, 1H, J=2.8, J=11.0Hz, H '-5b), 2.95 (d, 1H, J=6.0Hz, 2 '-OH), 0.91 (s, 9H, t-Bu), 0.05 (s, 3H, SiCH ₃), 0.00 (s, 3H, SiCH ₃).

Step H:5 '-O-(4,4 '-dimethoxytrityl)-2 '-O-(tert-butyl dimethylsilyl)-1 '-(5-nitroindoline base)-D-riboside-3 '-O-cyanoethyl-N, N-di-isopropyl phosphoramidate (110)

With 2-cyanoethyl-N, N-di-isopropyl torak acid amides (153mg, 0.646mmol) join 5 '-O-(4,4 '-dimethoxytrityl)-3 '-O-(tert-butyl dimethylsilyl)-1-(5-nitroindoline base)-D-β-riboside 108 (230mg, 0.323mmol), diisopropylethylamine (306uL, 1.78mmol) and DMAP (10mg) in the solution of exsiccant methylene dichloride (3mL), and at room temperature stir 4-6h in the argon gas.Reaction mixture is condensed into thick residue, and it is added to in the saturated silicagel column of the hexane solution of 2% triethylamine, and obtains the pure compound of amorphous solid shape 110 (250mg, 85% with hexane-ethyl acetate (2:1) wash-out.

³¹P-NMR(CDCl3，400MHz)：δ 149.54(s)，146.57(s).Anal.Cald ofC ₅₀H ₆₅N ₄O ₉PSi：924.43.Found：947.43[M+Na]+。

2 '-hydroxyl or the 3 '-hydroxyl solid carrier of step I:5 '-O-(4,4 '-dimethoxytrityl)-1-(5-nitroindoline base)-D-riboside (111).

Succinyl oxide is added in 5 '-O-(4,4 '-dimethoxytrityl)-1-(5-nitroindoline base)-2 '-OTBDMS (108) of D-β-riboside (109) or the solution of mixture in dry methylene chloride of 3 '-O-TBDMS and DMAP.Reaction mixture is at room temperature stirred 6h in the argon gas.Add another part of succinyl oxide and DMAP, and stir 16h altogether.Mixture is condensed into thick residue, it is dissolved in the ethyl acetate (50ml), with citric acid (400mg/20ml), salt water washing, and dry (Na ₂SO ₄).Organic layer is condensed into thick succsinic acid nucleosides.It is directly used in subsequent reactions need not purifying.

With succsinic acid nucleosides, DMAP, DTNP and Ph ₃P at room temperature stirs 20min[Nucleosideand nucleotides, and 1996,15 (4), 879-888.].Add lcaa-CPG then, and under uniform temp, stir 45min.Filter out solid, and use CH ₃CN, methylene dichloride and ether washing.The drying solid carrier, end-blocking and washing obtain solid carrier under standard procedure.

It is synthetic that controlled glass carrier that comprises the nitroindoline base that obtains thus and phosphoramidate are used in the standard oligonucleotide of the above-mentioned oligonucleotide that is used for comprising natural base.

The vitro test of embodiment 3:siRNA

Suppress the ability that influenza virus is duplicated in external human clone or mouse body build-in test iRNA reagent.The transfection of iRNA reagent in cell, for example, is made by transfection or electroporation to act on certain hour (for example, 24 hours) on cell, and determine the level of infection by forming plaque or elisa assay.For several influenza genes recombinant expressed in mammalian host cell, test rna i reagent is for the inhibition of expression of target gene, to replenish these direct analyses.

Virus and clone

Influenza virus A/PR/8/34 (PR8), hypotype H1N1 is obtained by Charles River Laboratories (ATCC # VR-1469).A/WSN/33 (WSN), hypotype H1N1 is by Thomas Chambers, University of Kentucky, Lexington, KY.USA (sees Castrucci, M.R., Deng .J Virol.1992,66:4647), or Dr.Peter Palese, Mount Sinai School of Medicine New YorkCity, NY, USA (see WO 04/028471) locate to obtain.(Tompkins, S.M. wait Proc.Natl.Acad.Sci.2004,101:8682) to the virus original seed in 34 ℃ of breeding 48-72h (PR8) or 37 ℃ breeding 24h (WSN) in the allantoic cavity of the egg that has the embryo.

Mdck cell is by U.S. typical case DSMZ (American Type CultureCollection) (ATCC, Rockville MD, USA; ATCC # CCL-34) locate to obtain, and in the MEM that contains 8% heat-inactivated fetal bovine serum (FBS), 2mM L-glutaminate, 1mM Sodium.alpha.-ketopropionate, 1.5g/L sodium bicarbonate and non-essential amino acid in 37 ℃ at 5% CO ₂/ 95% air atmosphere is growth down.

Vero E6 cercopithecus aethiops renal epithelial cell is by ATCC (Rockville MD, USA, ATCC #CRL-1586) locates to obtain, and in the additional DMEM of 4.5g/l D-glucose, 2mM L-glutaminate, 110mg/l Sodium.alpha.-ketopropionate, 10% foetal calf serum (Hyclone, Cat # 30070.03) and 0.1% penicillin/streptomycin in 37 ℃ at 5% CO ₂/ 95% air atmosphere is growth down.

The Cos-7 African green monkey kidney cell is by German microorganism and cell culture preservation center (GermanCollection of Microorganisms and Cell Cultures) (DSMZ, Braunschweig, Germany, DSMZ # ACC 60) locates to obtain, and at Dulbecco ' s MEM, 10% foetal calf serum, 2mM L-glutaminate, 1.2 μ g/ml sodium bicarbonates, 100u penicillin/100 μ g/ml Streptomycin sulphate (Biochrom AG, Berlin, Germany) middle growth.

Embodiment 3.1: plaque forms and detects

Cell cultures, siRNA transfection and virus infection.

Transfection the day before yesterday, with mdck cell with 7.5 x 10 ⁴Cells/well 0.5ml growth medium kind is in the 24-orifice plate.In siRNA transfection that day, mdck cell 80% merges.Before the transfection, add the 0.25ml growth medium to cell.

Before adding cell, with 1.5ml (50 μ l/ hole) Optimem I (Invitrogen) and 90 μ l (3 μ l/ hole) Lipofectamin 2000 (Invitrogen), the amount of enough 24-orifice plates transfection is mixed also in 2ml Sarstedt pipe and was hatched 10-15 minute under the room temperature.The siRNA that is dissolved in the appropriate amount of annealing buffer is joined in Optimem/lipofectamine 2000 mixtures, obtain the final concentration of needs, mix, and at room temperature hatched again 15-25 minute.Then, according to the indication of test design to each hole Dropwise 50 μ lsiRNA/ reagent complex.Then slowly swing plate guaranteeing thorough mixing, and in 37 ℃ at 5%CO ₂Hatched under/95% air atmosphere 14 hours.

Then, transfection media is slowly siphoned away, once with 0.25-0.5ml PBS washed cell, and add the MEM solution of 100 μ l different concns PR8 to each hole, in 37 ℃ hatch 1-2 hour after, add 0.5ml and cover substratum (MEM, 20mM HEPES, 0.075% NaHCO ₃, 2mM glutamine, 0.6% agarose, 0.5 μ g/ml TPCK-trypsinase), and in incubator with flat board in 37 ℃ at 5%CO ₂Hatched under/95% air atmosphere 48 hours.As described below then flat board is fixed, all kinds of diseases and ailments poison plaque is carried out immunostaining.

Immunostaining and virus are quantitatively

The PBS flushing port is used in host-infection in 48 hours, cell fixation in 10% formalin of neutral buffered 45 minutes.Then the hole is used infiltration damping fluid (1 x PBS, 2% FBS, 0.5% Saponin/TSM, 0.1% sodiumazide) sealing 15 minutes under the room temperature, and add the solution that 125 μ l contain the 0.5 μ g/ml mouse-anti biotin labeled antibody MAB8258B of influenza A (Chemicon).At room temperature hatch 1hr then, remove not combination and antibody 2 times with the PBS flushing port, and the PBS solution of 1 μ g/ml horseradish peroxidase (HRP) the conjugated streptavidin (Vector Laboratories) of 125 μ l is added each hole, flat board is hatched 45min, and it is inferior to give a baby a bath on the third day after its birth with PBS.Every hole adds 200 μ l tmb substrate (VectorLaboratories #SK-4400)/holes.Then in the dark, hatched 5-10 minute under the room temperature, stop colorimetric reaction, bleed off water, dry flat board with distilled water.With the counter-rotating opticmicroscope that 4X amplifies painted influenza plaque is counted.Plaque forms active and only uses Lipofectamin cells transfected (simulation process) relatively, and remains infection rate with [(plaque in handling cell forms activity)/(plaque in the simulation process cell forms activity)] x 100=%

Embodiment 3.2:ELISA detects:

Transfection the day before yesterday, with MDCK or Vero cell with 10 ⁴Cells/well 0.1ml growth medium kind is in the 96-orifice plate.In siRNA transfection that day, cell 80% merges.Before the transfection, add 44 μ l growth mediums to cell.

With 1.08ml (9 μ l/ hole) Optimem I (Invitrogen) and 42 μ l (0.35 μ l/ hole) Lipofectamin 2000 (Invitrogen), the amount of enough 96-orifice plates transfection is mixed also in 2ml Sarstedt pipe and was hatched 10-15 minute under the room temperature.The siRNA that is dissolved in the appropriate amount of annealing buffer is joined in Optimem/lipofectamine 2000 mixtures, obtain the final concentration of needs, mix, and at room temperature hatched again 15-25 minute.Then, the indication according to test design drips 10 μ lsiRNA/ reagent complex to each hole.Slowly swing plate and was hatched under the 5%CO2/95% air atmosphere 14 hours in 37 ℃ guaranteeing thorough mixing then.

Then, with the PBS washed cell once, infect with the MEM that 50 μ l have the PR8 influenza virus in every hole, hatched 1-2 hour.Then, with PBS washing flat board once, add and have the tryptic 200 μ l MEM of 0.25/0.5 μ g/ml (MDCK/VERO).Infect two days later, with 15min in the dull and stereotyped stuck-at-0% buffered formalin.Wash cell with PBS, at room temperature seal 15min, add 50 μ l and contain 0.5 μ g/ml biotin labeled anti-influenza A monoclonal antibody MAB8258B (Chemicon)/hole with the sealing damping fluid.At room temperature hatch 1hr,, and add the sealing buffered soln that 1 μ g/ml AP-conjugated streptavidin (Vector Laboratories) is contained in 50 μ l/ holes with PBS flushing port 2 times.Hatch 45min and, add the pNPP substrate solution of every hole 100 μ l with after the PBS washing 3 times.Flat board is at room temperature developed in the dark, and read at 405nm.

The inhibition that embodiment 3.3:siRNA is recombinant expressed to the influenza target gene

By GENEART (Regensburg, Germany), the consensus sequence (consulting table 5) of synthetic MP (SEQ ID NO:1453), NP (SEQ ID NO:1454), PA (SEQ ID NO:1455), PB1 (SEQ ID NO:1456) and PB2 (SEQ ID NO:1457), and be cloned in the GENEART standard vector.(both is from NEBn via AsiSI and NotI with MP and PA, Frankfurt, Germany) the site subclone is to psiCheck-2 (Promega, Mannheim, Germany) in, NP, (PB1) and PB2 then via XhoI and NotI, be created in the construct that the influenza gene is arranged between terminator codon and the Renilla luciferase polyA signal thus.Carry out end sequencing by GATC Biotech (Konstanz, Germany), confirm correct clone.

Transfection

With 1.5 x 10 ⁴Individual cells/well is on white with clear bottom 96 orifice plates (Greiner Bio-One GmbH, Frickenhausen, Germany) of Cos-7 cell inoculation in 75 μ l growth mediums.Behind the inoculating cell, with Lipofectamine2000 (Invitrogen) transfection of 50ng plasmid/hole, as hereinafter described to siRNAs, plasmid is diluted to final volume 12.5 μ l/ holes with Opti-MEM immediately, and whole flat board is made mastermix.

Behind the plasmid transfection 4 hours, carry out the siRNA transfection, quadruplicate.For each hole, the Lipofectamine2000 (Invitrogen GmbH, Karlsruhe, Germany) of 0.5 μ l and 12 μ l Opti-MEM (Invitrogen) are mixed together, and incubation 15 minutes at room temperature.For siRNA concentration is reached in the 100 μ l transfection volumes be 50nM, 5 μ MsiRNA in every hole with 1 μ l mixes with 11.5 μ l Opti-MEM, and Lipofectamine2000-Opti-MEM make up and incubation 15 minutes once more at room temperature.Between incubation period, from cell, remove growth medium, and replace with the fresh culture in 75 μ l/ holes.The siRNA-Lipofectamine2000 mixture is added in the cell fully, and at 37 ℃, 5% CO ₂Condition under in the moistening incubator (Heraeus GmbH, Hanau, Germany) incubation cell 24 hours.

Come harvested cell by substratum with from the Dual-Glo luciferase substrate of Dual-Glo LuciferaseAssay System (Promega, Mannheim, Germany) by the mixture that 1:1 forms by removing growth medium and adding 150 μ l.According to the rules that the relevant Dual-Glo luciferase of manufacturers detects, carry out luciferase and detect, and in Victor-Light 1420 luminescent counters (Perkin Elmer, Rodgau-J ü gesheim, Germany), measure fluorescence.At each value that obtains with Photinus pyralis LUC, will be with the value stdn of Renilla luciferase acquisition.To be made as 100% (at neomycin resistance gene) with the value that no specific siRNA obtains, the value that the siRNAs that uses at the influenza gene obtains will be carried out stdn.

The effective siRNA that filters out further identifies by dose response curve.According to above-mentioned rules, the dose response curve under the following siRNA concentration of drawing: 100nM, 25nM, 6.3nM, 1.6nM, 400pM, 100pM, 24pM, 6pM, 1.5pM, 380fM.By parameterized curve fitting (parametrized curve fitting), utilize the XLfit program, determine IC ₅₀Value.

Table 5: the viral consensus sequence that is cloned into influenza gene M P (SEQ ID NO:1453), NP (SEQ ID NO:1454), PA (SEQ ID NO:1455), PB1 (SEQ ID NO:1456) and the PB2 (SEQ ID NO:1457) of Cos-7 cell

MP:obtain the M1 from gi|13383290|gb|AB049165| influenza A virus (A/ parakeet/Chiba/1/97 (H9N2)) film ion channel, stromatin; M2, ATGAGTCTTC TAACCGAGGT CGAAACGTAC GTTCTCTCTA TCATCCCGTC AGGCCCCCTC 60 AAAGCCGAGA TCGCGCAGAG ACTTGAAGAT GTCTTTGCAG AGAAGAACAC AGATCTCGAG 120 GCTCTCATGG AATGGCTAAA GACAAGACCA ATCCTGTCAC CTCTGACTAA GGGGATTTTA 180 GGGTTTGTGT TCACGCTCAC CGTGCCCAGT GAGCGAGGAC TGCAGCGTAG ACGCTTTGTC 240 CAGAATGCCC TAAATGGGAA TGGAGACCCA AACAACATGG ACAGGGCAGT TAAACTATAC 300 AAGAAGCTGA AGAGGGAAAT AACATTCCAT GGGGCTAAGG AAGTTGCACT CAGTTACTCT 360 GCTGGTGCAC TTGCCAGTTG CATGGGTCTC ATATACAACC GGATGGGAAC AGTGACCACA 420 GAAGTGGCTC TTGGCCTAGT GTGTGCCACT TGTGAGCAGA TTGCAGATTC ACAACATCGG 480 TCCCACAGGC AGATGGCGAC TACCACCAAC CCACTAATCA GACATGAGAA CAGAATGGTG 540 CTGGCCAGCA CTACAGCTAA GGCTATGGAG CAGATGGCTG GATCAAGTGA GCAGGCAGCG 600 GAAGCCATGG AAGTCGCAAG TCAGGCTAGG CAGATGGTGC AGGCAATGAG GACAATTGGG 660 ACTCATCCTA GCTCCAGTGC AGGTCTAAAA GATAATCTTC TTGAAAATTT GCAGGCCTAC 720 CAGAAACGAA TGGGGGTGCA GATGCAGCGA TTCAAGTGAT CCTCTCGTTG TTGCAGCAAG 780 TATCATTGGG ATCTTGCACT TGATATTGTG GATTCTTGAT CGTCTTTTCT TCAAATGCAT 840 TTATCGTCGC CTTAAATACG GTTTGAAAAG AGGGCCTTCT ACGGAAGGAG TACCTGAGTC 900 TATGAGGGAA GAGTATCGAC AGGAACAGCA GAGTGCTGTG GATGTTGACG ATGGTCATTT 960 TGTCAACATA GAGCTGGAGT AA 982 SEQ ID NO：1453

NP：H5N1 gi|14326108|AF370122| ( A///3/97 ( H5N1 ) ) , CTCACTGAGT GACATCAAAA TCATGGCGTC TCAAGGCACC AAACGATCTT ATGAACAGAT 60 GGAAACTGGT GGAGAACGCC AGAATGCTAC TGAGATCAGA GCATCTGTTG GAAGAATGGT 120 TGGTGGAGTT GGGAGGTTTT ATATACAGAT GTGCACTGAA CTCAAACTCA GCGACTATGA 180 AGGAAGGCTG ATTCAGAACA GCATAACAAT AGAGAGAATG GTTGTCTCTG CATTTGATGA 240 AAGGAGGAAC AAATACCTGG AAGAACATCC CAGTGCGGGG AAGGACCCAA AGAAAACTGG 300 AGGTCCAATC TACCGAAGAA GAGACGGGAA ATGGGTGAGA GAGCTGATTC TGTATGACAA 360 AGAGGAGATC AGGAGAATTT GGCGTCAAGC GAACAATGGA GAAGATGCAA CTGCTGGTCT 420 CACTCACCTG ATGATCTGGC ATTCCAATCT AAATGATGCC ACATACCAGA GAACAAGAGC 480 TCTCGTGCGT ACTGGGATGG ACCCTAGAAT GTGCTCTCTG ATGCAAGGAT CAACTCTCCC 540 GAGGAGATCT GGAGCTGCTG GTGCGGCAGT AAAGGGAGTC GGAACTATGG TGATGGAACT 600 AATTCGGATG ATAAAGCGAG GGATTAACGA TCGGAATTTC TGGAGAGGTG AAAATGGGCG 660 AAGAACAAGG ATTGCATATG AGAGAATGTG CAACATTCTC AAAGGGAAAT TCCAAACAGC 720

AGCACAAAGA GCAATGATGG ATCAGGTACG GGAAAGCAGA AATCCTGGGA ATGCTGAGAT 780 CGAAGATCTC ATATTTCTGG CACGGTCTGC ACTCATCCTG AGAGGATCAG TGGCCCACAA 840 GTCCTGCTTG CCTGCTTGTG TGTACGGGCT TGCCGTGGCC AGTGGATATG ACTTTGAGAG 900 AGAAGGGTAC TCTCTGGTCG GGATTGATCC TTTCCGTCTG CTGCAAAACA GCCAGGTCTT 960 TAGTCTAATT AGACCAAATG AGAATCCAGC ACATAAAAGT CAATTGGTGT GGATGGCATG 1020 CCATTCTGCA GCATTTGAAG ATCTGAGAGT CTCAAGCTTC ATCAGAGGGA CAAGAGTGGC 1080 CCCAAGGGGA CAACTATCTA CTAGAGGAGT ACAAATTGCT TCAAATGAGA ACATGGAAAC 1140 AATGGACTCC AGCACTCTTG AACTGAGAAG CAGATATTGG GCTATAAGGA CCAGGAGTGG 1200 AGGAAACACC AACCAGCAGA GAGCATCTGC AGGACAAATC AGTGTGCAGC CTACTTTCTC 1260 GGTACAGAGA AATCTTCCCT TCGAAAGAGC GACCATTATG GCGGCATTCA CAGGGAATAC 1320 AGAGGGCAGA ACATCTGACA TGAGGACTGA AATCATAAGG ATGATGGAAA GCTCCAGACC 1380 AGAAGATGTG TCTTTCCAGG GGCGGGGAGT CTTCGAGCTC TCGGACGAAA AGGCAACGAA 1440 CCCGATCGTG CCTTCCTTTG ACATGAGTAA TGAAGGATCT TATTTCTTCG GAGACAATGC 1500 AGAGGAGTAT GACAATTGAA G 1521 SEQ ID NO：1454

PA：H5N1gi|47156500|AY585473| ( A///35/2001 ( H5N1 ) ) ( PA ) mRNA, ATGGAAGACT TTGTGCGACA ATGCTTCAAT CCAATGATTG TCGAGCTTGC GGAAAAGGCA 60 ATGAAAGAAT ATGGGGAAGA TCCGAAAATC GAAACGAACA AATTTGCAGC AATATGCACA 120 CACTTAGAAG TCTGTTTCAT GTATTCAGAT TTTCACTTTA TTGATGAACG GGGCGAATCA 180 ATAATTGTAG AATCTGGCGA TCCGAATGCA TTATTGAAAC ACCGATTTGA AATAATTGAA 240 GGAAGAGACC GAACAATGGC CTGGACAGTG GTGAATAGTA TCTGCAACAC CACAGGAGTT 300 GAGAAACCTA AATTTCTCCC AGATTTGTAT GACTACAAAG AGAACCGATT CATTGAAATT 360 GGAGTGACAC GGAGGGAAGT TCATATATAC TATCTAGAGA AAGCCAACAA GATAAAATCC 420 GAGAAGACAC ACATTCACAT ATTCTCATTC ACTGGGGAGG AAATGGCCAC CAAAGCGGAC 480 TACACCCTTG ATGAAGAGAG CAGGGCAAGA ATCAAAACCA GGCTGTTCAC CATAAGGCAG 540 GAAATGGCCA GTAGGGGTCT ATGGGATTCC TTTCGTCAGT CCGAGAGAGG CGAAGAGACA 600 ATTGAAGAAA GATTTGAAAT CACAGGAACC ATGCGCAGGC TTGCCGACCA AAGTCTCCCA 660 CCGAACTTCT CCAGCCTTGA AAACTTTAGA GCCTATGTGG ATGGATTCGA ACCGAACGGC 720 TGCATTGAGG GCAAGCTTTC TCAAATGTCA AAAGAAGTGA ACGCCAGAAT TGAGCCATTT 780 CTGAAGACAA CACCACGCCC TCTCAGATTA CCTGATGGGC CTCCCTGCTC TCAGCGGTCG 840 AAGTTCTTGC TGATGGATGC CCTTAAATTA AGCATCGAAG ACCCGAGTCA TGAGGGGGAG 900 GGGATACCGC TATATGATGC AATCAAATGC ATGAAAACAT TTTTCGGCTG GAAAGAGCCC 960 AACATCGTAA AACCACATGA AAAAGGCATA AACCCCAATT ACCTCCTGGC TTGGAAGCAA 1020 GTGCTGGCAG AACTCCAAGA TATTGAAAAT GAGGAGAAAA TCCCAAAAAC AAAGAACATG 1080 AAGAAAACAA GCCAATTGAA GTGGGCACTC GGTGAGAACA TGGCACCAGA GAAAGTAGAC 1140 TTTGAGGATT GCAAAGATGT TAGCGATCTA AGACAGTATG ACAGTGATGA ACCAGAGCCT 1200 AGATCACTAG CAAGCTGGAT CCAGAGTGAA TTCAACAAGG CATGTGAATT GACAGATTCG 1260 AGTTGGATTG AACTTGATGA AATAGGGGAA GACGTTGCTC CAATTGAGCA CATTGCAAGT 1320 ATGAGAAGGA ACTATTTCAC AGCGGAAGTA TCCCATTGCA GGGCCACTGA ATACATAATG 1380 AAGGGGGTGT ACATAAACAC AGCTCTGTTG AATGCATCCT GTGCAGCCAT GGATGACTTT 1440 CAACTGATTC CAATGATAAG CAAATGCAGA ACCAAAGAAG GAAGACGGAA AACTAACCTG 1500 TATGGATTCA TTATAAAAGG AAGATCCCAT TTGAGGAATG ATACCGATGT GGTAAACTTT 1560 GTGAGTATGG AATTCTCTCT TACTGACCCG AGGCTGGAGC CACACAAGTG GGAAAAGTAC 1620 TGTGTTCTCG AGATAGGAGA CATGCTCCTA CGGACTGCAA TAGGCCAAGT TTCAAGGCCC 1680 ATGTTCCTGT ATGTGAGAAC CAATGGAACC TCCAAGATCA AAATGAAATG GGGAATGGAG 1740 ATGAGGCGAT GCCTTCTTCA ATCCCTTCAA CAGATTGAGA GCATGATTGA GGCCGAGTCT 1800

TCTGTCAAAG AGAAAGACAT GACCAAAGAA TTCTTTGAAA ACAAATCAGA AACATGGCCA 1860 ATTGGAGAGT CACCCAAAGG AGTGGAGGAA GGCTCCATCG GGAAGGTGTG CAGAACCTTA 1920 CTGGCGAAAT CTGTGTTCAA CAGTCTATAT GCATCTCCAC AACTCGAGGG GTTTTCAGCT 1980 GAATCAAGAA AATTGCTTCT CATTGTTCAG GCACTTAGGG ACAACCTGGA ACCTGGGACC 2040 TTCGATCTTG GAGGGCTATA TGAAGCAATT GAGGAGTGCC TGATTAATGA TCCCTGGGTT 2100 TTGCTTAATG CGTCTTGGTT CAACTCCTTC CTCACACATG CACTGAAATA GTT 2153 SEQ ID NO：1455

PB1：H5N1gi|58531084|AB166860| ( A///7/2004 ( H5N1 ) ) 1PB1, ATGGATGTCA ATCCGACTTT ACTTTTCTTG AAAGTACCAG TGCAAAATGC TATAAGTACC 60 ACATTCCCTT ATACTGGAGA CCCTCCATAC AGCCATGGAA CAGGGACAGG ATACACCATG 120 GACACAGTCA ACAGAACACA CCAATATTCA GAAAAGGGGA AGTGGACAAC AAACACAGAG 180 ACTGGAGCAC CCCAACTCAA CCCGATTGAT GGACCACTAC CTGAGGATAA TGAGCCCTGT 240 GGGTATGCAC AAACAGATTG TGTATTGGAA GCAATGGCTT TCCTTGAAGA ATCCCACCCA 300 GGGATCTTTG AAAACTCGTG TCTTGAAACG ATGGAAATTG TTCAACAAAC AAGAGTGGAT 360 AAACTGACCC AAGGTCGCCA GACCTATGAC TGGACATTGA ATAGAAACCA ACCGGCTGCA 420 ACTGCTTTGG CCAACACTAT AGAAATCTTC AGATCGAACG GTCTAACAGC CAATGAATCG 480 GGACGGCTAA TAGATTTCCT CAAGGATGTG ATGGAATCAA TGGATAAGGA AGAAATGGAG 540 ATAACAACAC ATTTCCAGAG AAAGAGAAGA GTGAGGGACA ACATGACCAA GAAAATGGTC 600 ACACAAAGAA CAATAGGGAA GAAAAAACAA AGGCTGAACA AAAAGAGCTA CCTGATAAGA 660 GCACTGACAC TGAACACAAT GACAAAAGAT GCAGAAAGAG GCAAATTGAA GAGGCGAGCA 720 ATTGCAACAC CCGGAATGCA AATCAGAGGA TTCGTGTACT TTGTTGAAAC ACTAGCGAGG 780 AGTATCTGTG AGAAACTTGA GCAATCTGGA CTCCCAGTCG GAGGGAATGA GAAGAAGGCT 840 AAATTGGCAA ACGTCGTGAG GAAGATGATG ACTAACTCAC AAGATACTGA ACTCTCCTTT 900 ACAATTACTG GAGACAATAC CAAATGGAAT GAGAATCAGA ATCCTAGGAT GTTTCTGGCA 960 ATGATAACGT ACATCACAAG GAACCAGCCA GAATGGTTTC GGAATGTCTT AAGCATTGCC 1020 CCTATAATGT TCTCAAACAA AATGGCGAGA TTAGGAAAAG GATACATGTT CGAAAGTAAG 1080 AGCATGAAGT TACGAACACA AATACCAGCA GAAATGCTTG CAAACATTGA TCTCAAATAC 1140 TTCAATGAAT TAACGAAAAA GAAAATTGAG AAAATAAGAC CTCTATTAAT AGATGGTACA 1200 GCCTCATTGA GCCCTGGAAT GATGATGGGC ATGTTCAACA TGCTGAGTAC AGTCCTAGGA 1260 GTCTCAATCC TGAATCTTGG ACAGAAAAGG TACACCAAAA CCACATATTG GTGGGACGGA 1320 CTCCAATCCT CTGATGATTT CGCTCTCATC GTAAATGCAC CGAATCATGA GGGAATACAA 1380 GCAGGAGTGG ATAGGTTTTA TAGGACTTGT AAACTAGTTG GAATCAATAT GAGCAAGAAG 1440 AAGTCTTACA TAAATCGGAC AGGGACATTT GAATTCACGA GCTTTTTCTA CCGCTATGGA 1500 TTTGTAGCCA ATTTCAGTAT GGAGCTGCCC AGTTTTGGAG TGTCTGGAAT TAATGAATCG 1560 GCCGACATGA GCATTGGTGT TACAGTGATA AAGAACAATA TGATAAACAA CGACCTTGGG 1620 CCAGCAACAG CTCAGATGGC TCTTCAGCTA TTCATCAAGG ACTACAGATA CACATACCGA 1680 TGCCACAGAG GGGATACGCA AATCCAAACG AGGAGATCAT TCGAGCTGAA GAAGCTGTGG 1740 GAGCAAACCC GTTCAAAGGC AGGACTGTTG GTTTCAGATG GAGGACCAAA TCTATACAAT 1800 ATCCGAAATC TCCATATTCC TGAGGTCTGC TTAAAATGGG AATTGATGGA TGAAGATTAC 1860 CAGGGCAGAC TGTGTAATCC TCTGAATCCG TTCGTCAGCC ATAAGGAAAT TGAATCTGTC 1920 AACAATGCTG TAGTAATGCC AGCTCATGGC CCGGCCAAAA GCGTGGAATA TGATGCCGTT 1980 GCAACTACAC ATTCATGGAT TCCTAAAAGG AATCGTTCCA TTCTCAATAC GAGTCAAAGG 2040 GGAATTCTTG AGGATGAACA GATGTACCAG AAGTGCTGCA ATCTATTCGA GAAATTCTTC 2100 CCCAGCAGTT CATATCGGAG GCCAGTTGGA ATTTCCAGCA TGGTGGAGGC CATGGTGTCT 2160 AGGGCCCGAA TTGACGCACG AATTGATTTC GAGTCTGGAA GGATTAAGAA AGAAGAGTTT 2220 GCTGAGATCA TGAAGATCTG TTCCACCATT GAAGAGCTCA GACGGCAAAA ATAG 2274 SEQ ID NO：1456

PB2: derive from the viral consensus sequence of H5N1gi|19697859|AY059525| influenza A virus (A/ duck/Hong Kong/2986.1/2000 (H5N1)) fragment 1 polymerase (PB2) gene, the part coded sequence. ATGGAGAGAA TAAAAGAATT AAGAGATCTA ATGTCGCAGT CTCGCACTCG CGAGATACTA 60 ACAAAAACCA CTGTGGACCA TATGGCCATA ATCAAGAAAT ACACATCAGG AAGACAAGAG 120 AAGAACCCTG CTCTCAGAAT GAAATGGATG ATGGCAATGA AATATCCAAT CACAGCAGAC 180

AAGAGAATAA TAGAGATGAT TCCTGAAAGG AATGAACAAG GGCAGACGCT TTGGAGCAAG 240 ACAAATGATG CTGGATCGGA CAGGGTGATG GTGTCTCCCC TAGCTGTAAC TTGGTGGAAT 300 AGGAATGGGC CGACGACAAG TGCAGTCTAT TATCCAAAGG TTTACAAAAC ATACTTTGAG 360 AAGGTTGAAA GGTTAAAACA TGGAACCTTC GGTCCCGTTC ATTTCCGAAA CCAAATTAAA 420 ATACGCCGCC GAGTTGATAT AAATCCTGGC CATGCAGATC TCAATGCTAA AGAAGCACAA 480 GATGTCATCA TGGAGGTCGT TTTCCCAAAT GAAGTGGGAG CTAGAATATT GACATCAGAG 540 TCGCAATTGA CAATAACGAA AGAAAAGAAA GAAGAGCTCC AGGATTGTAA GATTGCTCCT 600 TTAATGGTTG CATACATGTT GGAAAGGGAA CTGGTCCGCA AAACCAGATT CCTACCGGTA 660 GCAGGCGGAA CAAGCAGTGT GTACATTGAG GTATTGCATT TGACTCAAGG GACCTGCTGG 720 GAACAGATGT ACACTCCAGG CGGAGAAGTG AGAAATGACG ATGTTATCCA GAGTATGATC 780 ATCGCTGCCA GAAACATTGT TAGGAGAGCA ACGGTATCAG CGGATCCACT GGCATCACTG 840 CTGGAGATGT GTCACAGCAC ACAAATTGGT GGGATAAGGA TGGTGGACAT CCTTAGGCAA 900 AATCCAACTG AGGAACAAGC TGTGGATATA TGCAAAGCAG CAATGGGTTT GAGGATCAGT 960 TCATCCTTTA GCTTTGGAGG CTTCACTTTC AAAAGAACAA GTGGAACATC CGTCAAGAAG 1020 GAAGAGGAAG TGCTTACAGG CAACCTCCAA ACATTGAAAA TAAGAGTACA TGAGGGGTAT 1080 GAGGAATTCA CAATGGTTGG GCGGAGGGCA ACAGCTATCC TGAGGAAAGC AACTAGAAGG 1140 CTGATTCAGT TGATAGTAAG TGGAAGAGAC GAACAATCAA TCGCTGAGGC AATCATTGTA 1200 GCAATGGTGT TCTCACAGGA GGATTGCATG ATAAAGGCAG TCCGAGGCGA TCTGAATTTC 1260 GTAAACAGAG CAAACCAAAG ATTAAACCCC ATGCATCAAC TCCTGAGACA TTTTCAAAAG 1320 GATGCAAAAG TGCTATTTCA GAATTGGGGA ATTGAACCCA TTGATAATGT CATGGGGATG 1380 ATCGGAATAT TACCTGACAT GACTCCCAGC ACAGAAATGT CACTGAGAGG AGTAAGAGTT 1440 AGTAAAATGG GAGTGGATGA ATATTCCAGC ACTGAGAGAG TAGTTGTAAG TATTGACCGT 1500 TTCTTAAGGG TTCGAGATCA GCGGGGGAAC GTACTCTTAT CTCCCGAAGA GGTCAGCGAA 1560 ACACAGGGAA CAGAGAAATT GGCAATAACA TATTCATCAT CAATGATGTG GGAAATCAAC 1620 GGTCCTGAGT CAGTGCTTGT TAACACCTAT CAATGGATCA TCAGAAACTG GGAGACTGTG 1680 AAGATTCAAT GGTCTCAAGA CCCCACGATG CTGTACAATA AGATGGAGTT TGAACCGTTC 1740 CAATCCTTGG TACCTAAAGC TGCCAGAGGT CAATACAGTG GATTTGTGAG AACACTATTC 1800 CAACAAATGC GTGACGTACT GGGGACATTT GATACTGTCC AGATAATAAA GCTGCTACCA 1860 TTTGCAGCAG CCCCACCGGA GCAGAGCAGA ATGCAGTTTT CTTCTCTAAC TGTGAATGTG 1920 AGAGGCTCAG GAATGAGAAT ACTTGTAAGG GGCAATTCCC CTGTGTTCAA CTACAATAAG 1980 GCAACCAAAA GGCTTACCGT TCTTGGAAAG GACGCAGGTG CATTAACAGA GGATCCAGAT 2040 GAGGGAACAG CCGGAGTGGA ATCTGCAGTA CTGAGGGGAT TCCTAATTCT AGGCAAGGAG 2100 GACAAAAGAT ATGGACCAGC ATTGAGCATC AATGAACTGA GCAATCTTGC GAAAGGGGAG 2160 AAAGCTAATG TGCTGATAGG GCAAGGAGAC GTGGTGTTGG TAATGAAACG GAAACGGGAC 2220 TCTAGCATAC TTACTGACAG CCAGACAGCG ACCAAAAGAA TTCGGATGGC CATCAATTAG 2280 SEQ ID NO：1457

Table 1A, C, D and E have enumerated for the exemplary agents that the present invention selectes the target gene of the identifier of duplex, sense strand and antisense strand sequence, reagent and above-mentioned test gained result.Table 1B and H have enumerated for containing the selected exemplary agents of modification property nucleic acid group in order to degrade stable, the duplex identifier, the duplex identifier of corresponding unmodified sequence, the sequence of sense strand and antisense strand and the target gene of reagent, wherein all pyrimidine bases that contain Nucleotide comprise 2 ＇-O-methyl group on sense strand, and all pyrimidine bases that contain Nucleotide in 5 ＇-ca-3 ＇ or 5 ＇-ua-3 ＇ sequence background comprise 2 ＇-O-methyl group in antisense strand, except antisense strand does not wherein comprise those reagent of Nucleotide in 5 ＇-ca-3 ＇ or 5 ＇-ua-3 ＇ sequence background, wherein the uridine in all 5 ＇-ug-3 ＇ sequence background is Nucleotide (for example, the AL-DP-2295 that 2 ＇-O-methyl is modified in antisense strand, AL-DP-2301 and AL-DP-2302).For some particularly preferred RNAi reagent of the present invention, the concentration when table 2 listed for 50% maximum the inhibition, described maximum inhibition is to measure according to the dose response in the Cos-7 cell through transforming the expression of influenza gene.

Table 6

Influenza A virus stromatin (Matrix Protein, MP) used sequence in the analysis

AY180470 influenza A virus strain A/ quail/Nanchang/12-340/2000 (H1N1) stromatin (M) gene, part encoding sequence (partialcds).

AY633213 influenza A virus (A/ wild duck (mallard)/alberta province (Alberta)/211/98 (H1N1)) stromatin (M) gene, encoding sequence (complete cds) completely.

The stromatin mRNA of AY664487 influenza A virus (A/ wild duck/alberta is economized/119/98 (H1N1)) non-functional, partial sequence.

M55476 influenza A virus stromatin (M1) gene, encoding sequence and M2 albumen (M2) gene, encoding sequence completely completely.

M55479 influenza A virus stromatin (M1) gene, encoding sequence and M2 albumen (M2) gene, encoding sequence completely completely.

M55480 influenza A virus stromatin (M1) gene, encoding sequence and M2 albumen (M2) gene, encoding sequence completely completely.

M63528 influenza A virus (A/ turkey/Minnesota State/166/81 (H1N1)) membranin M1 and membranin M2 gene, encoding sequence completely.

U49119 influenza A virus matrix prote m1 and M2 (M) gene, encoding sequence completely.

The M of Z26859 influenza A virus stromatin and M2 gene

The M of Z26860 influenza A virus stromatin and M2 gene

AY422021 influenza A virus (A/ duck/Hokkaido/95/01 (H2N2)) stromatin 1 (M) gene, the part encoding sequence.

The RNA fragment 7 of M12699 bird influenza A/wild duck/NY/6750/78 coding M1 and M2, encoding sequence completely.

AF213915 influenza A virus (A/ chicken/Italy/5945/95 (H3N2)) fragment 7 stromatins (M) gene, the part encoding sequence.

AY180498 influenza A virus strain A/ chicken/Nanchang/3-120/2001 (H3N2) stromatin (M) gene, the part encoding sequence.

The stromatin mRNA of AY664458 influenza A virus (A/ turnstone (ruddy turnstone)/Delaware/142/99 (H3N2)) non-functional, partial sequence.

AY769614 influenza A virus (A/ turkey/Ohio/313053/04 (H3N2)) stromatin gene, the part encoding sequence.

AY779257 influenza A virus (A/ turkey/North Carolina State/12344/03 (H3N2)) stromatin 2 (M) gene, the part encoding sequence; With stromatin 1 (M) gene, encoding sequence completely.

AY779258 influenza A virus (A/ turkey/Minnesota State/764-2/03 (H3N2)) stromatin 2 (M) gene, the part encoding sequence; With stromatin 1 (M) gene, encoding sequence completely.

AY862623 influenza A virus (A/ chicken/Korea S/S6/03 (H3N2)) stromatin (M) gene, encoding sequence completely.

AY862624 influenza A virus (A/ duck/Korea S/S7/03 (H3N2)) stromatin (M) gene, encoding sequence completely.

AY862625 influenza A virus (A/ duck/Korea S/S8/03 (H3N2)) stromatin (M) gene, encoding sequence completely.

AY862626 influenza A virus (A/ duck/Korea S/S9/03 (H3N2)) stromatin (M) gene, encoding sequence completely.

AY862627 influenza A virus (A/ duck/Korea S/S10/03 (H3N2)) stromatin (M) gene, encoding sequence completely.

AY862628 influenza A virus (A/ dove/Korea S/S11/03 (H3N2)) stromatin (M) gene, encoding sequence completely.

Z26858 influenza A virus stromatin M and M2 gene

M1 and M2 gene, the encoding sequence completely of AB166865 influenza A virus (A/ chicken/mountain pass/7/2004 (H5N1)) stromatin and film ionic channel.

M2, M1 gene, the encoding sequence completely of AB188819 influenza A virus (A/ chicken/big (Ojta)/8/2004 (H5N1) of dividing) film ionic channel 2 and stromatin 1.

AF509043 influenza A virus (A/ chicken/Hong Kong/FY150/01 (H5N1)) M1 albumen (M1) gene, encoding sequence completely.

AF509044 influenza A virus (A/ pheasant/Hong Kong/FY155/01 (H5N1)) M1 albumen (M1) gene, encoding sequence completely.

AF509045 influenza A virus (A/ Gallus Domesticus (silky chicken)/Hong Kong/SF189/01 (H5N1)) M1 albumen (M1) gene, encoding sequence completely.

AF509046 influenza A virus (A/ quail/Hong Kong/SF203/01 (H5N1)) M1 albumen (M1) gene, encoding sequence completely.

AF509047 influenza A virus (A/ pigeon (pigeon)/Hong Kong/SF215/01 (H5N1)) M1 albumen (M1) gene, encoding sequence completely.

AF509048 influenza A virus (A/ chicken/Hong Kong/SF219/01 (H5N1)) M1 albumen (M1) gene, encoding sequence completely.

AF509049 influenza A virus (A/ chicken/Hong Kong/715.5/01 (H5N1)) M1 albumen (M1) gene, encoding sequence completely.

AF509050 influenza A virus (A/ chicken/Hong Kong/751.1/01 (H5N1)) M1 albumen (M1) gene, encoding sequence completely.

AF509051 influenza A virus (A/ chicken/Hong Kong/822.1/01 (H5N1)) M1 albumen (M1) gene, encoding sequence completely.

AF509052 influenza A virus (A/ chicken/Hong Kong/829.2/01 (H5N1)) M1 albumen (M1) gene, encoding sequence completely.

AF509053 influenza A virus (A/ chicken/Hong Kong/830.2/01 (H5N1)) M1 albumen (M1) gene, encoding sequence completely.

AF509054 influenza A virus (A/ chicken/Hong Kong/858.3/01 (H5N1)) M1 albumen (M1) gene, encoding sequence completely.

AF509055 influenza A virus (A/ chicken/Hong Kong/866.3/01 (H5N1)) M1 albumen (M1) gene, encoding sequence completely.

AF509056 influenza A virus (A/ chicken/Hong Kong/867.1/01 (H5N1)) M1 albumen (M1) gene, encoding sequence completely.

AF509057 influenza A virus (A/ chicken/Hong Kong/879.1/01 (H5N1)) M1 albumen (M1) gene, encoding sequence completely.

AF509058 influenza A virus (A/ chicken/Hong Kong/873.3/01 (H5N1)) M1 albumen (M1) gene, encoding sequence completely.

AF509059 influenza A virus (A/ chicken/Hong Kong/876.1/01 (H5N1)) M1 albumen (M1) gene, encoding sequence completely.

AF509060 influenza A virus (A/ chicken/Hong Kong/891.1/01 (H5N1)) M1 albumen (M1) gene, encoding sequence completely.

AF509061 influenza A virus (A/ chicken/Hong Kong/893.2/01 (H5N1)) M1 albumen (M1) gene, encoding sequence completely.

AF509062 influenza A virus (A/ goose/Hong Kong/76.1/01 (H5N1)) M1 albumen (M1) gene, encoding sequence completely.

AF509063 influenza A virus (A/ goose/Hong Kong/ww100/01 (H5N1)) M1 albumen (M1) gene, encoding sequence completely.

AF509064 influenza A virus (A/ duck/Hong Kong/573.4/01 (H5N1)) M1 albumen (M1) gene, encoding sequence completely.

AF509065 influenza A virus (A/ duck/Hong Kong/646.3/01 (H5N1)) M1 albumen (M1) gene, encoding sequence completely.

AY059506 influenza A virus (A/ goose/Hong Kong/ww26/2000 (H5N1)) fragment 7 stromatins (M) gene, the part encoding sequence.

AY059507 influenza A virus (A/ goose/Hong Kong/ww28/2000 (H5N1)) fragment 7 stromatins (M) gene, the part encoding sequence.

AY059508 influenza A virus (A/ duck/Hong Kong/ww381/2000 (H5N1)) fragment 7 stromatins (M) gene, the part encoding sequence.

AY059509 influenza A virus (A/ duck/Hong Kong/ww461/2000 (H5N1)) fragment 7 stromatins (M) gene, the part encoding sequence.

AY059510 influenza A virus (A/ goose/Hong Kong/ww491/2000 (H5N1)) fragment 7 stromatins (M) gene, the part encoding sequence.

AY059511 influenza A virus (A/ duck/Hong Kong/2986.1/2000 (H5N1)) fragment 7 stromatins (M) gene, the part encoding sequence.

AY059512 influenza A virus (A/ goose/Hong Kong/3014.8/2000 (H5N1)) fragment 7 stromatins (M) gene, the part encoding sequence.

AY075029 influenza A virus (A/ chicken/Hong Kong/317.5/2001 (H5N1)) stromatin 1 and stromatin 2 (M) gene, encoding sequence completely.

AY075035 influenza A virus (A/ duck/Hong Kong/380.5/2001 (H5N1)) stromatin 1 and stromatin 2 (M) gene, encoding sequence completely.

AY221530 influenza A virus (A/ chicken/Hong Kong/NT873.3/01-MB (H5N1)) stromatin (M) gene, encoding sequence completely.

AY221531 influenza A virus (A/ chicken/Hong Kong/NT873.3/01 (H5N1)) stromatin (M) gene, encoding sequence completely.

AY221532 influenza A virus (A/ chicken/Hong Kong/FY150/01-MB (H5N1)) stromatin (M) gene, encoding sequence completely.

AY221533 influenza A virus (A/ chicken/Hong Kong/FY150/01 (H5N1)) stromatin (M) gene, encoding sequence completely.

AY221534 influenza A virus (A/ pheasant/Hong Kong/FY155/01-MB (H5N1)) stromatin (M) gene, encoding sequence completely.

AY221535 influenza A virus (A/ pheasant/Hong Kong/FY155/01 (H5N1)) stromatin (M) gene, encoding sequence completely.

AY221536 influenza A virus (A/ chicken/Hong Kong/YU822.2/01-MB (H5N1)) stromatin (M) gene, encoding sequence completely.

AY221537 influenza A virus (A/ chicken/Hong Kong/YU822.2/01 (H5N1)) stromatin (M) gene, encoding sequence completely.

AY221538 influenza A virus (A/ chicken/Hong Kong/YU562/01 (H5N1)) stromatin (M) gene, encoding sequence completely.

AY518361 influenza A virus (A/ duck/China/E319-2/03 (H5N1)) film ionic channel M2 and matrix prote m1 (M) gene, encoding sequence completely.

AY575895 influenza A virus (A/Gs/ Hong Kong/739.2/02 (H5N1)) stromatin (M) gene, encoding sequence completely.

AY575896 influenza A virus (A/Eg/ Hong Kong/757.3/02 (H5N1)) stromatin (M) gene, the part encoding sequence.

AY575897 influenza A virus (A/G.H/ Hong Kong/793.1/02 (H5N1)) stromatin (M) gene, the part encoding sequence.

AY575898 influenza A virus (A/Dk/ Hong Kong/821/02 (H5N1)) stromatin (M) gene, the part encoding sequence.

AY575899 influenza A virus (A/Ck/ Hong Kong/31.4/02 (H5N1)) stromatin (M) gene, encoding sequence completely.

AY575900 influenza A virus (A/Ck/ Hong Kong/61.9/02 (H5N1)) stromatin (M) gene, encoding sequence completely.

AY575901 influenza A virus (A/Ck/ Hong Kong/YU777/02 (H5N1)) stromatin (M) gene, encoding sequence completely.

AY575902 influenza A virus (A/Ck/ Hong Kong/96.1/02 (H5N1)) stromatin (M) gene, encoding sequence completely.

AY575903 influenza A virus (A/Ck/ Hong Kong/409.1/02 (H5N1)) stromatin (M) gene, encoding sequence completely.

AY575904 influenza A virus (A/Ph/ Hong Kong/sv674.15/02 (H5N1)) stromatin (M) gene, encoding sequence completely.

AY585378 influenza A virus (A/ duck/Fujian/01/2002 (H5N1)) stromatin mRNA, encoding sequence completely.

AY585379 influenza A virus (A/ duck/Fujian/13/2002 (H5N1)) stromatin mRNA, encoding sequence completely.

AY585380 influenza A virus (A/ duck/Fujian/17/2001 (H5N1)) stromatin mRNA, encoding sequence completely.

AY585381 influenza A virus (A/ duck/Fujian/19/2000 (H5N1)) stromatin mRNA, encoding sequence completely.

AY585382 influenza A virus (A/ duck/Guangdong/01/2001 (H5N1)) stromatin mRNA, encoding sequence completely.

AY585383 influenza A virus (A/ duck/Guangdong/07/2000 (H5N1)) stromatin mRNA, encoding sequence completely.

AY585384 influenza A virus (A/ duck/Guangdong/12/2000 (H5N1)) stromatin mRNA, encoding sequence completely.

AY585385 influenza A virus (A/ duck/Guangdong/22/2002 (H5N1)) stromatin mRNA, encoding sequence completely.

AY585386 influenza A virus (A/ duck/Guangdong/40/2000 (H5N1)) stromatin mRNA, encoding sequence completely.

AY585387 influenza A virus (A/ duck/Guangxi/07/1999 (H5N1)) stromatin mRNA, encoding sequence completely.

AY585388 influenza A virus (A/ duck/Guangxi/22/2001 (H5N1)) stromatin mRNA, the part encoding sequence.

AY585389 influenza A virus (A/ duck/Guangxi/35/2001 (H5N1)) stromatin mRNA, encoding sequence completely.

AY585390 influenza A virus (A/ duck/Guangxi/53/2002 (H5N1)) stromatin mRNA, encoding sequence completely.

AY585391 influenza A virus (A/ duck/Shanghai/08/2001 (H5N1)) stromatin mRNA, encoding sequence completely.

AY585392 influenza A virus (A/ duck/Shanghai/13/2001 (H5N1)) stromatin mRNA, encoding sequence completely.

AY585393 influenza A virus (A/ duck/Shanghai/35/2002 (H5N1)) stromatin mRNA, encoding sequence completely.

AY585394 influenza A virus (A/ duck/Shanghai/37/2002 (H5N1)) stromatin mRNA, encoding sequence completely.

AY585395 influenza A virus (A/ duck/Shanghai/38/2001 (H5N1)) stromatin mRNA, encoding sequence completely.

AY585396 influenza A virus (A/ duck/Zhejiang/11/2000 (H5N1)) stromatin mRNA, encoding sequence completely.

AY585397 influenza A virus (A/ duck/Zhejiang/52/2000 (H5N1)) stromatin mRNA, encoding sequence completely.

AY585398 influenza A virus (A/ duck/Guangxi/50/2001 (H5N1)) stromatin mRNA, encoding sequence completely.

AY590578 influenza A virus (A/ chicken/Buddhist system (Nakorn-Patom)/Thailand/CU-K2/2004 (H5N1)) stromatin M2 and matrix prote m1 (M) gene, part and encoding sequence completely.

AY609315 influenza A virus (A/ chicken/Guangdong/174/04 (H5N1)) fragment 7, sequence completely.

AY651374 influenza A virus (A/Ck/ Indonesia/BL/2003 (H5N1)) film ionic channel 2 (M) gene, the part encoding sequence; With stromatin 1 (M) gene, encoding sequence completely.

AY651375 influenza A virus (A/Dk/ Indonesia/MS/2004 (H5N1)) film ionic channel 2 and stromatin 1 (M) gene, encoding sequence completely.

AY651376 influenza A virus (A/Ck/ Indonesia/PA/2003 (H5N1)) film ionic channel 2 and stromatin 1 (M) gene, encoding sequence completely.

AY651377 influenza A virus (A/Ck/ Indonesia/2A/2003 (H5N1)) film ionic channel 2 and stromatin 1 (M) gene, encoding sequence completely.

AY651378 influenza A virus (A/Ck/ Indonesia/4/2004 (H5N1)) film ionic channel 2 and stromatin 1 (M) gene, encoding sequence completely.

AY651379 influenza A virus (A/Ck/ Indonesia/5/2004 (H5N1)) film ionic channel 2 and stromatin 1 (M) gene, encoding sequence completely.

AY651380 influenza A virus (A/Ck/ Thailand/1/2004 (H5N1)) film ionic channel 2 and stromatin 1 (M) gene, encoding sequence completely.

AY651381 influenza A virus (A/Ck/ Thailand/73/2004 (H5N1)) film ionic channel 2 (M) gene, the part encoding sequence; With stromatin 1 (M) gene, encoding sequence completely.

AY651382 influenza A virus (A/Ck/ Thailand/9.1/2004 (H5N1)) film ionic channel 2 (M) gene, the part encoding sequence; With stromatin 1 (M) gene, encoding sequence completely.

AY651383 influenza A virus (A/Qa/ Thailand/57/2004 (H5N1)) film ionic channel 2 and stromatin 1 (M) gene, encoding sequence completely.

AY651384 influenza A virus (A/ bird (bird)/Thailand/3.1/2004 (H5N1)) film ionic channel 2 (M) gene, the part encoding sequence; With stromatin 1 (M) gene, encoding sequence completely.

AY651385 influenza A virus (A/Dk/ Thailand/71.1/2004 (H5N1)) film ionic channel 2 (M) gene, the part encoding sequence; And stromatin 1 (M) gene, encoding sequence completely.

AY651386 influenza A virus (A/Gs/ Thailand/79/2004 (H5N1)) film ionic channel 2 (M) gene, the part encoding sequence; And stromatin 1 (M) gene, encoding sequence completely.

AY651391 influenza A virus (A/Ck/ Vietnam/33/2004 (H5N1)) film ionic channel 2 and stromatin 1 (M) gene, encoding sequence completely.

AY651392 influenza A virus (A/Ck/ Vietnam/35/2004 (H5N1)) film ionic channel 2 and stromatin 1 (M) gene, encoding sequence completely.

AY651393 influenza A virus (A/Ck/ Vietnam/36/2004 (H5N1)) film ionic channel 2 (M) gene, the part encoding sequence; And stromatin 1 (M) gene, encoding sequence completely.

AY651394 influenza A virus (A/Ck/ Vietnam/37/2004 (H5N1)) film ionic channel 2 and stromatin 1 (M) gene, encoding sequence completely.

AY651395 influenza A virus (A/Ck/ Vietnam/38/2004 (H5N1)) film ionic channel 2 and stromatin 1 (M) gene, encoding sequence completely.

AY651396 influenza A virus (A/Ck/ Vietnam/39/2004 (H5N1)) film ionic channel 2 (M) gene, the part encoding sequence; And stromatin 1 (M) gene, encoding sequence completely.

AY651397 influenza A virus (A/Ck/ Vietnam/C57/2004 (H5N1)) film ionic channel 2 (M) gene, the part encoding sequence; And stromatin 1 (M) gene, encoding sequence completely.

AY651398 influenza A virus (A/Dk/ Vietnam/11/2004 (H5N1)) film ionic channel 2 and stromatin 1 (M) gene, encoding sequence completely.

AY651399 influenza A virus (A/Gf/ Hong Kong/38/2002 (H5N1)) film ionic channel 2 and stromatin 1 (M) gene, the part encoding sequence.

AY651400 influenza A virus (A/Ck/ Hong Kong/31.2/2002 (H5N1)) film ionic channel 2 and stromatin 1 (M) gene, encoding sequence completely.

AY651401 influenza A virus (A/Ck/ Hong Kong/37.4/2002 (H5N1)) film ionic channel 2 and stromatin 1 (M) gene, encoding sequence completely.

AY651402 influenza A virus (A/SCk/ Hong Kong/YU100/2002 (H5N1)) film ionic channel 2 and stromatin 1 (M) gene, encoding sequence completely.

AY651403 influenza A virus (A/Ck/ Hong Kong/YU22/2002 (H5N1)) film ionic channel 2 and stromatin 1 (M) gene, encoding sequence completely.

AY651404 influenza A virus (A/Ck/ Hong Kong/3176.3/2002 (H5N1)) film ionic channel 2 and stromatin 1 (M) gene, the part encoding sequence.

AY651405 influenza A virus (A/Ck/ Hong Kong/3169.1/2002 (H5N1)) stromatin 1 and film ionic channel 2 (M) gene, the part encoding sequence.

AY651406 influenza A virus (A/Ck/ Hong Kong/FY157/2003 (H5N1)) film ionic channel 2 and stromatin 1 (M) gene, encoding sequence completely.

AY651407 influenza A virus (A/Ck/ Hong Kong/YU324/2003 (H5N1)) film ionic channel 2 and stromatin 1 (M) gene, encoding sequence completely.

AY651408 influenza A virus (A/Ck/ Hong Kong/2133.1/2003 (H5N1)) film ionic channel 2 and stromatin 1 (M) gene, the part encoding sequence.

AY651409 influenza A virus (A/Ck/ Hong Kong/NT93/2003 (H5N1)) film ionic channel 2 (M) gene, the part encoding sequence; With stromatin 1 (M) gene, encoding sequence completely.

AY651410 influenza A virus (A/Ck/ Hong Kong/SSP141/2003 (H5N1)) film ionic channel 2 (M) gene, the part encoding sequence; With stromatin 1 (M) gene, encoding sequence completely.

AY651411 influenza A virus (A/Ck/ Hong Kong/WF157/2003 (H5N1)) film ionic channel 2 and stromatin 1 (M) gene, encoding sequence completely.

AY651412 influenza A virus (A/ falcon/Hong Kong/D0028/2004 (H5N1)) film ionic channel 2 (M) gene, the part encoding sequence; With stromatin 1 (M) gene, encoding sequence completely.

AY651413 influenza A virus (the red mouth of A/ gull/Hong Kong/12.1/2003 (H5N1)) film ionic channel 2 and stromatin 1 (M) gene, encoding sequence completely.

AY651414 influenza A virus (A/ heron/Hong Kong/861.1/2002 (H5N1)) film ionic channel 2 and stromatin 1 (M) gene, encoding sequence completely.

AY651415 influenza A virus (A/ rock dove/Hong Kong/862.7/2002 (H5N1)) film ionic channel 2 and stromatin 1 (M) gene, encoding sequence completely.

AY651416 influenza A virus (A/ sets sparrow (tree sparrow)/Hong Kong/864/2002 (H5N1)) stromatin 1 and film ionic channel 2 (M) gene, the part encoding sequence.

AY651417 influenza A virus (A/teal/ China/2978.1/2002 (H5N1)) film ionic channel 2 and stromatin 1 (M) gene, the part encoding sequence.

AY651418 influenza A virus (A/Dk/ Honduras (HN)/5806/2003 (H5N1)) film ionic channel 2 (M) gene, the part encoding sequence; With stromatin 1 (M) gene, encoding sequence completely.

AY651419 influenza A virus (A/Dk/ST/4003/2003 (H5N1)) film ionic channel 2 (M) gene, the part encoding sequence; With stromatin 1 (M) gene, encoding sequence completely.

AY651420 influenza A virus (A/Ck/ST/4231/2003 (H5N1)) film ionic channel 2 (M) gene, the part encoding sequence; With stromatin 1 (M) gene, encoding sequence completely.

AY651421 influenza A virus (A/Dk/YN/6255/2003 (H5N1)) film ionic channel 2 (M) gene, the part encoding sequence; With stromatin 1 (M) gene, encoding sequence completely.

AY651422 influenza A virus (A/Dk/YN/6445/2003 (H5N1)) film ionic channel 2 (M) gene, the part encoding sequence; With stromatin 1 (M) gene, encoding sequence completely.

AY651423 influenza A virus (A/Ck/Y/374/2004 (H5N1)) film ionic channel 2 (M) gene, the part encoding sequence; With stromatin 1 (M) gene, encoding sequence completely.

AY651424 influenza A virus (A/Dk/HN/101/2004 (H5N1)) film ionic channel 2 (M) gene, the part encoding sequence; With stromatin 1 (M) gene, encoding sequence completely.

AY651425 influenza A virus (A/Dk/HN/303/2004 (H5N1)) film ionic channel 2 (M) gene, the part encoding sequence; With stromatin 1 (M) gene, encoding sequence completely.

AY651426 influenza A virus (A/Ph/ST/44/2004 (H5N1)) film ionic channel 2 (M) gene, the part encoding sequence; With stromatin 1 (M) gene, encoding sequence completely.

AY651427 influenza A virus (A/Ck/YN/115/2004 (H5N1)) film ionic channel 2 (M) gene, the part encoding sequence; With stromatin 1 (M) gene, encoding sequence completely.

AY653194 influenza A virus (A/ chicken/Jilin/9/2004 (H5N1)) fragment 7, sequence completely.

AY676045 influenza A virus strain (A/ duck/Hong Kong/821/02 (H5N1)) membranin (M) gene, encoding sequence completely.

AY676046 influenza A virus strain (A/ egression (egret)/Hong Kong/757.2/03 (H5N1)) membranin (M) gene, encoding sequence completely.

AY676047 influenza A virus strain (A/ chicken/Korea S/ES/03 (H5N1)) membranin (M) gene, encoding sequence completely.

AY676048 influenza A virus strain (A/ duck/Korea S/ESD1/03 (H5N1)) membranin (M) gene, encoding sequence completely.

AY684709 influenza A virus (A/ chicken/Hubei/327/2004 (H5N1)) stromatin 2 (M2) and stromatin 1 (M1) gene s, encoding sequence completely.

AY737292 influenza A virus (A/ chicken/Guangdong/191/04 (H5N1)) fragment 7, sequence completely.

AY737298 influenza A virus (A/ chicken/Guangdong/178/04 (H5N1)) fragment 7, sequence completely.

AY737306 influenza A virus (A/ duck/Guangdong/173/04 (H5N1)) fragment 7, sequence completely.

AY770077 influenza A virus (A/ chicken/Hubei/489/2004 (H5N1)) stromatin 2 (M2) and stromatin 1 (M1) gene s, encoding sequence completely.

AY770998 influenza A virus (A/ chicken/big subtlety (Ayutthaya)/Thailand/CU-23/04 (H5N1)) stromatin gene, encoding sequence completely.

AY818145 influenza A virus (A/ chicken/Vietnam/C58/04 (H5N1)) matrix prote m1 gene, encoding sequence completely.

AY818146 influenza A virus (A/ quail/Vietnam/36/04 (H5N1)) matrix prote m1 gene, encoding sequence completely.

AY856865 influenza A virus (A/ duck/Shandong/093/2004 (H5N1)) fragment, sufficient sequence.

DQ055851 influenza A virus (A/ chicken/Yunnan/K001/2004 (H5N1)) matrix prote m1 gene, encoding sequence completely.

AB189048 influenza A virus (A/ chicken/capital of a country/3/2004 (H5N1)) film ionic channel M2, the M1 gene; M2, stromatin 1, encoding sequence completely.

AB189056 influenza A virus (A/ crow/capital of a country/53/2004 (H5N1)) film ionic channel M2, M1; M2, stromatin 1, encoding sequence completely.

AB189064 influenza A virus (A/ crow/Osaka/102/2004 (H5N1)) film ionic channel M2, the M1 gene; M2, stromatin 1, encoding sequence completely.

AF046082 influenza A virus (A/ chicken/Hong Kong/220/97 (H5N1)) stromatin 2 (M2) and stromatin 1 (M1) gene s, encoding sequence completely.

AF098560 influenza A virus (A/ chicken/Hong Kong/258/97 (H5N1)) M1 stromatin (M) and M2 stromatin (M) gene, the part encoding sequence.

AF098561 influenza A virus (A/ chicken/Hong Kong/y388/97 (H5N1)) M1 stromatin (M) and M2 stromatin (M) gene, the part encoding sequence.

AF098562 influenza A virus (A/ chicken/Hong Kong/728/97 (H5N1)) M1 stromatin (M) and M2 stromatin (M) gene, the part encoding sequence.

AF098563 influenza A virus (A/ chicken/Hong Kong/786/97 (H5N1)) M1 stromatin (M) and M2 stromatin (M) gene, the part encoding sequence.

AF098564 influenza A virus (A/ chicken/Hong Kong/915/97 (H5N1)) M1 stromatin (M) and M2 stromatin (M) gene, the part encoding sequence.

AF098566 influenza A virus (A/ duck/Hong Kong/p46/97 (H5N1)) M1 stromatin (M) and M2 stromatin (M) gene, the part encoding sequence.

AF098567 influenza A virus (A/ duck/Hong Kong/y283/97 (H5N1)) M1 stromatin (M) and M2 stromatin (M) gene, the part encoding sequence.

AF098568 influenza A virus (A/ goose/Hong Kong/w355/97 (H5N1)) M1 stromatin (M) and M2 stromatin (M) gene s, the part encoding sequence.

AF144306 influenza A virus (A/ goose/Guangdong/1/96 (H5N1)) matrix prote m1 and M2 (M) gene, or montage product, encoding sequence completely.

AF216711 influenza A virus (A/ environment (Environment)/Hong Kong/437-4/99 (H5N1)) stromatin 1 and stromatin 2 genes, encoding sequence completely.

AF216719 influenza A virus (A/ environment/Hong Kong/437-6/99 (H5N1)) stromatin 1 and stromatin gene, encoding sequence completely.

AF216727 influenza A virus (A/ environment/Hong Kong/437-8/99 (H5N1)) stromatin 1 and stromatin 2 gene s, encoding sequences completely.

AF216735 influenza A virus (A/ environment/Hong Kong/437-10/99 (H5N1)) stromatin 1 and stromatin 2 genes, encoding sequence completely.

AF359560 influenza A virus (A/ goose/Guangdong/3/97 (H5N1)) stromatin 1 and stromatin 2 (M) gene, encoding sequence completely.

AF398429 influenza A virus (A/ goose/Hong Kong/385.3/2000 (H5N1)) stromatin 1 (M) gene, the part encoding sequence.

AF398430 influenza A virus (A/ goose/Hong Kong/385.5/2000 (H5N1)) stromatin 1 (M) gene, the part encoding sequence.

AF468843 influenza A virus (A/ duck/Anyang (Anyang)/AVL-1/2001 (H5N1)) stromatin 1 and stromatin 2 genes, encoding sequence completely.

AF509040 influenza A virus (A/ chicken/Hong Kong/FY77/01 (H5N1)) M1 albumen (M1) gene, encoding sequence completely.

AF509041 influenza A virus (A/ chicken/Hong Kong/YU562/01 (H5N1)) M1 albumen (M1) gene, encoding sequence completely.

AF509042 influenza A virus (A/ chicken/Hong Kong/YU563/01 (H5N1)) M1 albumen (M1) gene, encoding sequence completely.

AF073180 influenza A virus (A/ chicken/New Jersey/15086-3/94 (H7N3NSA)) stromatin 1 (M1) and stromatin 2 (M2) gene, encoding sequence completely.

AF073197 influenza A virus (A/ turkey/Oregon/71 (H7N3NSB)) stromatin 1 (M1) and stromatin 2 (M2) gene, encoding sequence completely.

The stromatin mRNA of AY664433 influenza A virus (A/ turnstone/New Jersey/65/85 (H7N3)) non-functional, partial sequence.

AY677732 influenza A virus (A/ chicken/British Columbia/CN7-3/04 (H7N3)) stromatin 1 (M1) gene, encoding sequence completely.

AF073198 influenza A virus (A/ turkey/state of Colorado/13356/91 (H7N3NSA)) stromatin 1 (M1) and stromatin 2 (M2) gene, encoding sequence completely.

AF073200 influenza A virus (A/ quail/Arkansas State/16309-7/94 (H7N3NSA)) stromatin 1 (M1) and stromatin 2 (M2) gene, encoding sequence completely.

AF073201 influenza A virus (A/ turkey/Utah State/24721-10/95 (H7N3NSA)) stromatin 1 (M1) and stromatin 2 (M2) gene, encoding sequence completely.

AJ627492 influenza A virus (A/ turkey/Italy/214845/2002 (H7N3)) gene of membranin 1 and the gene of membranin 2, geneome RNA.

AJ627497 influenza A virus (A/ turkey/Italy/220158/2002 (H7N3)) gene of membranin 1 and the gene of albumen 2, geneome RNA.

AY241600 influenza A virus (A/ chicken/New York/12273-11/99 (H7N3)) stromatin 1 and stromatin 2 genes, encoding sequence completely.

AY241602 influenza A virus (A/ chicken/NY/14714-9/99 (H7N3)) stromatin 1 and stromatin 2 genes, encoding sequence completely.

AY241615 influenza A virus (A/ duck/NJ/117228-7/01 (H7N3)) stromatin 1 and stromatin 2 genes, encoding sequence completely.

AY241616 influenza A virus (A/ duck/PA/143585/01 (H7N3)) stromatin 1 and stromatin 2 genes, encoding sequence completely.

The AY300975 influenza A virus (the short neck duck (Blue-winged Teal) of the blue wing of A//TX/2/01 (H7N3) membranin (M) gene, encoding sequence completely.

AY303652 influenza A virus (A/ chicken/Chile/176822/02 (H7N3)) stromatin 1 and stromatin 2 genes, encoding sequence completely.

AY303653 influenza A virus (A/ chicken/Chile/4322/02 (H7N3)) stromatin 1 and stromatin 2 genes, encoding sequence completely.

AY303654 influenza A virus (A/ chicken/Chile/4957/02 (H7N3)) stromatin 1 gene, encoding sequence completely; With stromatin 2 genes, part encoding sequence.

AY303655 influenza A virus (A/ chicken/Chile/4968/02 (H7N3)) stromatin 1 and stromatin 2 genes, encoding sequence completely.

AY303656 influenza A virus (A/ chicken/Chile/4977/02 (H7N3)) stromatin 1 and stromatin 2 genes, encoding sequence completely.

AY303657 influenza A virus (A/ turkey/Chile/4418/02 (H7N3)) stromatin 1 gene, encoding sequence completely; With stromatin 2 genes, part encoding sequence.

AY586427 influenza A virus (A/ turkey/Italy/214845/02 (H7N3)) stromatin gene, the part encoding sequence.

AY586428 influenza A virus (A/ turkey/Italy/220158/2002 (H7N3)) stromatin gene, the part encoding sequence.

AY586429 influenza A virus (A/ wild duck/Italy/43/01 (H7N3)) stromatin gene, the part encoding sequence.

AY586430 influenza A virus (A/ wild duck/Italy/33/01 (H7N3)) stromatin gene, the part encoding sequence.

AY611525 influenza A virus (A/ chicken/British Columbia/04 (H7N3)) stromatin 2 (M) and stromatin 1 (M) gene, encoding sequence completely.

AY646079 influenza A virus (A/ chicken/British Columbia/GSC_human_B/04 (H7N3)) stromatin 2 and stromatin 1 (M) gene, encoding sequence completely.

AY648288 influenza A virus (A/GSC_ chicken _ B/ British Columbia/04 (H7N3)) stromatin 2 (M) and stromatin 1 (M) gene, encoding sequence completely.

AY650271 influenza A virus (A/GSC_ chicken/British Columbia/04 (H7N3)) stromatin 2 (M) and stromatin 1 (M) gene, encoding sequence completely.

The M1 gene of AJ619676 influenza A virus (A/ chicken/Germany/R28/03 (H7N7)) membranin 1, geneome RNA.

AY340086 influenza A virus (A/ Holland/124/03 (H7N7)) stromatin gene, the part encoding sequence.

AY340087 influenza A virus (A/ Holland/126/03 (H7N7)) stromatin gene, the part encoding sequence.

AY340088 influenza A virus (A/ Holland/127/03 (H7N7)) stromatin gene, the part encoding sequence.

AY340089 influenza A virus (A/ Holland/219/03 (H7N7)) stromatin gene, encoding sequence completely.

AY340090 influenza A virus (A/ Holland/33/03 (H7N7)) stromatin gene, encoding sequence completely.

AY340091 influenza A virus (A/ chicken/Holland/1/03 (H7N7)) stromatin gene, encoding sequence completely.

AY664468 influenza A virus (A/ turnstone/Delaware/134/99 (H7N7)) non-functional stromatin mRNA, partial sequence.

L37795 influenza A virus/chicken/Brescia/1902 (H7N7) stromatin (M1) gene and transmembrane protein (M2) gene, encoding sequence completely.

L37796 influenza A virus/FPV/ the nymph of the dragonfly (H7N7) stromatin (M1) gene and transmembrane protein (M2) gene, encoding sequence completely.

L37797 influenza A virus/FPV/Weybridge (H7N7) stromatin (M1) gene and transmembrane protein (M2) gene, encoding sequence completely.

M23917 influenza A/chicken/FPV/Weybridge (H7N7) M1 stromatin gene, encoding sequence completely.

M23921 influenza A virus A/ chicken/FPV/Weybridge (H7N7) M2 stromatin gene, encoding sequence completely.

M38299 influenza A/FPV/Weybridge (H7N7) matrix (M) albumen (fragment 7) gene, encoding sequence completely.

M63523 influenza A virus (A/ chicken/Victoria/1/85 (H7N7)) membranin M1 and membranin M2 gene, encoding sequence completely.

M63526 influenza A virus (the strain A/FPV/ nymph of the dragonfly/27 (H7N7)) membranin M1 and membranin M2 gene, encoding sequence completely.

AB049165 influenza A virus (A/ parakeet/Chiba/1/97 (H9N2)) film ionic channel, matrix prote m1, M2 gene, encoding sequence completely.

AB049166 influenza A virus (A/ parakeet/Narita Airport/92A/98 (H9N2)) film ionic channel, the M1 of stromatin, M2 gene, encoding sequence completely.

AF222671 influenza A virus (A/ Gallus Domesticus/Hong Kong/SF44/99 (H9N2)) fragment 7M1 (M1) gene, the part encoding sequence.

AF508684 influenza A virus (A/ ostrich/South Africa/9508103/95 (H9N2)) fragment 7 matrix prote m1s (M) gene, encoding sequence completely.

AF508685 influenza A virus (A/ chicken/Pakistan/4/99 (H9N2)) fragment 7 matrix prote m1s (M) gene, the part encoding sequence.

AF508686 influenza A virus (A/ chicken/Pakistan/5/99 (H9N2)) fragment 7 matrix prote m1s (M) gene, the part encoding sequence.

AF508687 influenza A virus (A/ chicken/Germany/R45/98 (H9N2)) fragment 7 matrix prote m1s (M) gene, encoding sequence completely.

AF508688 influenza A virus (A/ duck/Germany/113/95 (H9N2)) fragment 7 matrix prote m1s (M) gene, encoding sequence completely.

AF508689 influenza A virus (A/ chicken/Iran/11T/99 (H9N2)) fragment 7 matrix prote m1s (M) gene, the part encoding sequence.

AF508690 influenza A virus (A/ chicken/Saudi Arabia/532/99 (H9N2)) fragment 7 matrix prote m1s (M) gene, the part encoding sequence.

AF508691 influenza A virus (A/ pheasant/Ireland/PV18/97 (H9N2)) fragment 7 matrix prote m1s (M) gene, encoding sequence completely.

AF508692 influenza A virus (A/ chicken/Korea S/99029/99 (H9N2)) fragment 7 matrix prote m1s (M) gene, encoding sequence completely.

AF508693 influenza A virus (A/ chicken/Beijing/8/98 (H9N2)) fragment 7 matrix prote m1s (M) gene, encoding sequence completely.

AF508694 influenza A virus (A/ chicken/Guangdong/10/00 (H9N2)) fragment 7 matrix prote m1s (M) gene, encoding sequence completely.

AF508695 influenza A virus (A/ chicken/Guangdong/11/97 (H9N2)) fragment 7 matrix prote m1s (M) gene, encoding sequence completely.

AF508696 influenza A virus (A/ chicken/Heilungkiang/10/97 (H9N2)) fragment 7 matrix prote m1s (M) gene, encoding sequence completely.

AF508697 influenza A virus (A/ chicken/Henan/62/00 (H9N2)) fragment 7 matrix prote m1s (M) gene, encoding sequence completely.

AF508698 influenza A virus (A/ chicken/Ningxia/5/99 (H9N2)) fragment 7 matrix prote m1s (M) gene, encoding sequence completely.

AF508699 influenza A virus (A/ chicken/Sichuan/5/97 (H9N2)) fragment 7 matrix prote m1s (M) gene, the part encoding sequence.

AF508700 influenza A virus (A/ chicken/Shandong/6/96 (H9N2)) fragment 7 matrix prote m1s (M) gene, encoding sequence completely.

AF508701 influenza A virus (A/ chicken/Shijiazhuang/2/99 (H9N2)) fragment 7 matrix prote m1s (M) gene, the part encoding sequence.

AF508702 influenza A virus (A/ chicken/Shenzhen/9/97 (H9N2)) fragment 7 matrix prote m1s (M) gene, encoding sequence completely.

AF508703 influenza A virus (A/ duck/Nanjing/1/97 (H9N2)) fragment 7 matrix prote m1s (M) gene, encoding sequence completely.

AF508704 influenza A virus (A/ quail/Shanghai/8/96 (H9N2)) fragment 7 matrix prote m1s (M) gene, encoding sequence completely.

AF523482 influenza A virus (A/ duck/Shantou/1043/00 (H9N2)) stromatin (M) gene, encoding sequence completely.

AF523483 influenza A virus (A/ duck/Shantou/2134/00 (H9N2)) stromatin (M) gene, encoding sequence completely.

AF523484 influenza A virus (A/Wild duck/Shantou/4808/01 (H9N2)) stromatin (M) gene, encoding sequence completely.

AF523485 influenza A virus (A/ duck/Shantou/1042/00 (H9N2)) stromatin (M) gene, encoding sequence completely.

AF523486 influenza A virus (A/ duck/Shantou/2143/00 (H9N2)) stromatin (M) gene, encoding sequence completely.

AF523487 influenza A virus (A/ duck/Shantou/2144/00 (H9N2)) stromatin (M) gene, encoding sequence completely.

AF523488 influenza A virus (A/ duck/Shantou/1881/00 (H9N2)) stromatin (M) gene, encoding sequence completely.

AF523489 influenza A virus (A/ duck/Shantou/1796/00 (H9N2)) stromatin (M) gene, encoding sequence completely.

AF523490 influenza A virus (A/ duck/Shantou/2102/00 (H9N2)) stromatin (M) gene, encoding sequence completely.

AF523491 influenza A virus (A/ duck/Shantou/830/00 (H9N2)) stromatin (M) gene, encoding sequence completely.

AF523492 influenza A virus (A/ duck/Shantou/2088/01 (H9N2)) stromatin (M) gene, encoding sequence completely.

AF523493 influenza A virus (A/ duck/Shantou/1605/01 (H9N2)) stromatin (M) gene, encoding sequence completely.

AF523494 influenza A virus (A/ duck/Hong Kong/610/79 (H9N2)) stromatin (M) gene, encoding sequence completely.

AF523495 influenza A virus (A/ duck/Hong Kong/552/79 (H9N2)) stromatin (M) gene, encoding sequence completely.

AF523496 influenza A virus (A/ duck/Hong Kong/289/78 (H9N2)) stromatin (M) gene, encoding sequence completely.

AF523497 influenza A virus (A/ duck/Hong Kong/86/76 (H9N2)) stromatin (M) gene, encoding sequence completely.

AF523498 influenza A virus (A/ duck/Hong Kong/366/78 (H9N2)) stromatin (M) gene, the part encoding sequence.

AF536719 influenza A virus (A/ chicken/Beijing/1/95 (H9N2)) non-functional stromatin gene, partial sequence.

AF536720 influenza A virus (A/ chicken/Beijing/2/97 (H9N2)) non-functional stromatin gene, partial sequence.

AF536721 influenza A virus (A/ chicken/Beijing/3/99 (H9N2)) non-functional stromatin gene, partial sequence.

AF536722 influenza A virus (A/ chicken/Guangdong/97 (H9N2)) non-functional stromatin gene, partial sequence.

AF536723 influenza A virus (A/ chicken/Hebei/1/96 (H9N2)) non-functional stromatin gene, partial sequence.

AF536724 influenza A virus (A/ chicken/Hebei/2/98 (H9N2)) non-functional stromatin gene, partial sequence.

AF536725 influenza A virus (A/ chicken/Hebei/3/98 (H9N2)) non-functional stromatin gene, partial sequence.

AF536726 influenza A virus (A/ chicken/Henan/98 (H9N2)) non-functional stromatin gene e, partial sequence.

AF536727 influenza A virus (A/ chicken/Liaoning/99 (H9N2)) non-functional stromatin gene, partial sequence.

AF536728 influenza A virus (A/ chicken/Shandong/98 (H9N2)) non-functional stromatin gene, partial sequence.

The M1 gene of AJ291398 influenza A virus (A/ chicken/Pakistan/2/99 (H9N2)) stromatin 1 (exons 1) and the M2 gene of stromatin 2 (exons 1 and 2), geneome RNA

The stromatin m gene of AJ427865 influenza A virus (A/ quail/Hong Kong/FY298/00 (H9N2)) part, geneome RNA

AY180461 influenza A virus strain A/ pigeon/Nanchang/2-0461/2000 (H9N2) stromatin (M) gene, the part encoding sequence.

AY180462 influenza A virus strain A/ duck/Nanchang/11-290/2000 (H9N2) stromatin (M) gene, the part encoding sequence.

AY180463 influenza A virus strain A/ duck/Nanchang/11-197/2000 (H9N2) stromatin (M) gene, the part encoding sequence.

AY180464 influenza A virus strain A/ duck/Nanchang/11-392/2000 (H9N2) stromatin (M) gene, the part encoding sequence.

AY180477 influenza A virus strain A/ chicken/Nanchang/4-361/2001 (H9N2) stromatin (M) gene, the part encoding sequence.

AY180485 influenza A virus strain A/ pigeon/Nanchang/11-145/2000 (H9N2) stromatin (M) gene, the part encoding sequence.

AY180486 influenza A virus strain A/ duck/Nanchang/1-0070/2000 (H9N2) stromatin (M) gene, the part encoding sequence.

AY180489 influenza A virus strain A/ duck/Nanchang/10-389/2000 (H9N2) stromatin (M) gene, the part encoding sequence.

AY180490 influenza A virus strain A/ chicken/Nanchang/1-0016/2000 (H9N2) stromatin (M) gene, the part encoding sequence.

AY180492 influenza A virus strain A/ pigeon/Nanchang/7-058/2000 (H9N2) stromatin (M) gene, the part encoding sequence.

AY180495 influenza A virus strain A/ quail/Nanchang/2-0460/2000 (H9N2) stromatin (M) gene, the part encoding sequence.

AY180502 influenza A virus strain A/ chicken/Nanchang/4-010/2000 (H9N2) stromatin (M) gene, the part encoding sequence.

AY180504 influenza A virus strain A/ quail/Nanchang/4-040/2000 (H9N2) stromatin (M) gene, the part encoding sequence.

AY180506 influenza A virus strain A/ chicken/Nanchang/4-301/2001 (H9N2) stromatin (M) gene, the part encoding sequence.

AY180516 influenza A virus strain A/ duck/Nanchang/7-092/2000 (H9N2) stromatin (M) gene, the part encoding sequence.

AY180519 influenza A virus strain A/Wild duck/Nanchang/2-0480/2000 (H9N2) stromatin (M) gene, the part encoding sequence.

AY253755 influenza A virus (A/ chicken/Shanghai/F/98 (H9N2)) matrix prote m1 and film ionic channel M2 gene, encoding sequence completely.

AY496852 influenza A virus (A/ chicken/Mudanjiang/0823/2000 (H9N2)) stromatin (M1) mRNA, encoding sequence completely.

AY633165 influenza A virus (A/ wild duck/alberta is economized/17/91 (H9N2)) stromatin (M) gene, encoding sequence completely.

AY633277 influenza A virus (A/ wild duck/alberta is economized/321/88 (H9N2)) stromatin (M) gene, encoding sequence completely.

AY633293 influenza A virus (A/ wild duck/alberta is economized/11/91 (H9N2)) stromatin (M) gene, encoding sequence completely.

AY664464 influenza A virus (A/ bank bird (shorebird)/Delaware/276/99 (H9N2)) nonfunctional stromatin mRNA, partial sequence.

AY664679 influenza A virus (A/ chicken/Hong Kong/CSW153/03 (H9N2)) membranin M2 (M) gene, the part encoding sequence; With albumen M1 (M) gene, encoding sequence completely.

AY664680 influenza A virus (A/ chicken/Hong Kong/AP45/03 (H9N2)) membranin M2 (M) gene, the part encoding sequence; With membranin M1 (M) gene, encoding sequence completely.

AY664681 influenza A virus (A/ chicken/Hong Kong/BD90/03 (H9N2)) membranin M2 (M) gene, the part encoding sequence; With membranin M1 (M) gene, encoding sequence completely.

AY664682 influenza A virus (A/ chicken/Hong Kong/CSW291/03 (H9N2)) membranin M2 (M) gene, the part encoding sequence; With membranin M1 (M) gene, encoding sequence completely.

AY664683 influenza A virus (A/ chicken/Hong Kong/CSW304/03 (H9N2)) membranin M2 (M) and membranin M1 (M) gene, the part encoding sequence.

AY664684 influenza A virus (A/ chicken/Hong Kong/FY23/03 (H9N2)) membranin M2 (M) gene, the part encoding sequence; With membranin M1 (M) gene, encoding sequence completely.

AY664685 influenza A virus (A/ vulture galeeny (guineafowl)/Hong Kong/NT101/03 (H9N2)) membranin M2 (M) gene, the part encoding sequence; With membranin M1 (M) gene, encoding sequence completely.

AY664686 influenza A virus (A/ chicken/Hong Kong/NT142/03 (H9N2)) membranin M2 (M) gene, the part encoding sequence; With membranin M1 (M) gene, encoding sequence completely.

AY664687 influenza A virus (A/ chicken/Hong Kong/SF1/03 (H9N2)) membranin M2 (M) gene, the part encoding sequence; With membranin M1 (M) gene, encoding sequence completely.

AY664688 influenza A virus (A/ chicken/Hong Kong/SSP101/03 (H9N2)) membranin M2 (M) gene, the part encoding sequence; With membranin M1 (M) gene, encoding sequence completely.

AY664689 influenza A virus (A/ chicken/Hong Kong/TP38/03 (H9N2)) membranin M2 (M) gene, the part encoding sequence; With membranin M1 (M) gene, encoding sequence completely.

AY664690 influenza A virus (A/ chicken/Hong Kong/WF126/03 (H9N2)) membranin M2 (M) gene, the part encoding sequence; With membranin M1 (M) gene, encoding sequence completely.

AY664691 influenza A virus (A/ pigeon/Hong Kong/WF53/03 (H9N2)) membranin M2 (M) gene, the part encoding sequence; With membranin M1 (M) gene, encoding sequence completely.

AY664692 influenza A virus (A/ pheasant/Hong Kong/WF54/03 (H9N2)) membranin M2 (M) gene, the part encoding sequence; With membranin M1 (M) gene, encoding sequence completely.

AY664693 influenza A virus (A/ vulture galeeny/Hong Kong/NT184/03 (H9N2)) membranin M2 (M) gene, the part encoding sequence; With membranin M1 (M) gene, encoding sequence completely.

AY664694 influenza A virus (A/ chicken/Hong Kong/WF120/03 (H9N2)) membranin M2 (M) and membranin M1 (M) gene, the part encoding sequence.

AY664695 influenza A virus (A/ chicken/Hong Kong/NT366/03 (H9N2)) membranin M2 (M) gene, the part encoding sequence; With membranin M1 (M) gene, encoding sequence completely.

AY664696 influenza A virus (A/ chicken/Hong Kong/SSP418/03 (H9N2)) membranin M2 (M) gene, the part encoding sequence; With membranin M1 (M) gene, encoding sequence completely.

AY664697 influenza A virus (A/ chicken/Hong Kong/YU427/03 (H9N2)) membranin M2 (M) gene, the part encoding sequence; With membranin M1 (M) gene, encoding sequence completely.

AY800234 influenza A virus (A/ chicken/Korea S/S1/2003 (H9N2)) stromatin (M) gene, encoding sequence completely.

AY862614 influenza A virus (A/ Gallus Domesticus/Korea S/S3/03 (H9N2)) stromatin (M) gene, encoding sequence completely.

AY862615 influenza A virus (A/ chicken/Korea S/S4/03 (H9N2)) stromatin (M) gene, encoding sequence completely.

AY862616 influenza A virus (A/ chicken/Korea S/S5/03 (H9N2)) stromatin (M) gene, encoding sequence completely.

AY862617 influenza A virus (A/ chicken/Korea S/S12/03 (H9N2)) stromatin (M) gene, encoding sequence completely.

AY862618 influenza A virus (A/ duck/Korea S/S13/03 (H9N2)) stromatin (M) gene, encoding sequence completely.

AY862619 influenza A virus (A/ dove (dove)/Korea S/S14/03 (H9N2)) stromatin (M) gene, the part encoding sequence.

AY862620 influenza A virus (A/ chicken/Korea S/S15/03 (H9N2)) stromatin (M) gene, encoding sequence completely.

AY862621 influenza A virus (A/ chicken/Korea S/S16/03 (H9N2)) stromatin (M) gene, encoding sequence completely.

AY862622 influenza A virus (A/ chicken/Korea S/S18/03 (H9N2)) stromatin (M) gene, encoding sequence completely.

AF156458 influenza A virus (A/ chicken/Hong Kong/G9/97 (H9N2)) fragment 7 matrix prote m1s (M1) and stromatin M2 (M2) gene, encoding sequence completely.

AF156459 influenza A virus (A/ chicken/Hong Kong/G23/97 (H9N2)) fragment 7 matrix prote m1s (M1) and stromatin M2 (M2) gene, encoding sequence completely.

AF156460 influenza A virus (A/ pigeon/Hong Kong/Y233/97 (H9N2)) fragment 7 matrix prote m1s (M1) and stromatin M2 (M2) gene, encoding sequence completely.

AF156461 influenza A virus (A/ duck/Hong Kong/Y280/97 (H9N2)) fragment 7 matrix prote m1s (M1) and stromatin M2 (M2) gene, encoding sequence completely.

AF156462 influenza A virus (A/ duck/Hong Kong/Y439/97 (H9N2)) fragment 7 matrix prote m1s (M1) and stromatin M2 (M2) gene, encoding sequence completely.

AF156463 influenza A virus (A/ quail/Hong Kong/G1/97 (H9N2)) fragment 7 matrix prote m1s (M1) gene, encoding sequence completely; With stromatin M2 (M2) gene, part encoding sequence.

AF156464 influenza A virus (A/ chicken/Hong Kong/739/94 (H9N2)) fragment 7 matrix prote m1s (M1) gene, encoding sequence completely; With stromatin M2 (M2) gene, part encoding sequence.

AF156465 influenza A virus (A/ quail/Hong Kong/AF157/92 (H9N2)) fragment 7 matrix prote m1s (M1) gene, encoding sequence completely; With stromatin M2 (M2) gene, part encoding sequence.

AF156466 influenza A virus (A/ chicken/Beijing/1/94 (H9N2)) fragment 7 matrix prote m1s (M1) gene, encoding sequence completely; With stromatin M2 (M2) gene, part encoding sequence.

AF156467 influenza A virus (A/ chicken/Korea S/38349-p96323/96 (H9N2)) fragment 7 matrix prote m1s (M1) gene, encoding sequence completely; With stromatin M2 (M2) gene, part encoding sequence.

AF156468 influenza A virus (A/ chicken/Korea S/25232-96006/96 (H9N2)) fragment 7 matrix prote m1s (M1) gene, encoding sequence completely; With stromatin M2 (M2) gene, part encoding sequence.

AF156469 influenza A virus (A/ bank bird/Delaware/9/96 (H9N2)) fragment 7 matrix prote m1s (M1) gene, encoding sequence completely; With stromatin M2 (M2) gene, part encoding sequence.

AF156470 influenza A virus (A/ quail/Arkansas State/29209-1/93 (H9N2)) fragment 7 matrix prote m1s (M1) gene, encoding sequence completely; With stromatin M2 (M2) gene, part encoding sequence.

AF156471 influenza A virus (A/ turkey/California/189/66 (H9N2)) fragment 7 matrix prote m1s (M1) gene, encoding sequence completely; With stromatin M2 (M2) gene, part encoding sequence.

AF203788 influenza A virus (A/ chicken/Korea S/MS96/96 (H9N2)) stromatin 1mRNA, encoding sequence completely; With stromatin 2mRNA, the part encoding sequence.

AF222662 influenza A virus (A/ quail/Hong Kong/A17/99 (H9N2)) fragment 7M1 (M1) gene, the part encoding sequence.

AF222663 influenza A virus (A/ pigeon/Hong Kong/FY6/99 (H9N2)) fragment 7M1 (M1) gene, the part encoding sequence.

AF222664 influenza A virus (A/ chicken/Hong Kong/NT16/99 (H9N2)) fragment 7M1 (M1) gene, the part encoding sequence.

AF222665 influenza A virus (A/ quail/Hong Kong/SSP10/99 (H9N2)) fragment 7M1 (M1) gene, the part encoding sequence.

AF222666 influenza A virus (A/ pheasant/Hong Kong/SSP11/99 (H9N2)) fragment 7M1 (M1) gene, the part encoding sequence.

AF222667 influenza A virus (A/ chicken/Hong Kong/FY20/99 (H9N2)) fragment 7M1 (M1) gene, the part encoding sequence.

AF222668 influenza A virus (A/ chicken/Hong Kong/KC12/99 (H9N2)) fragment 7M1 (M1) gene, the part encoding sequence.

AF222669 influenza A virus (A/ quail/Hong Kong/NT28/99 (H9N2)) fragment 7M1 (M1) gene, the part encoding sequence.

AF222670 influenza A virus (A/ chicken/Hong Kong/SF2/99 (H9N2)) fragment 7M1 (M1) gene, the part encoding sequence.

Influenza A nucleocapsid protein (Nucleocapsid Protein, NP) used sequence in the analysis

AF156415 influenza A virus (A/ turkey/California/189/66 (H9N2)) fragment 5 nucleoprotein (NP) gene, the part encoding sequence.

AF523423 influenza A virus (A/ duck/Hong Kong/86/76 (H9N2)) nucleocapsid protein (NP) gene, encoding sequence completely.

AF523424 influenza A virus (A/ duck/Hong Kong/366/78 (H9N2)) nucleocapsid protein (NP) gene, the part encoding sequence.

AF523421 influenza A virus (A/ duck/Hong Kong/289/78 (H9N2)) nucleocapsid protein (NP) gene, encoding sequence completely.

AF523422 influenza A virus (A/ duck/Hong Kong/552/79 (H9N2)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY633279 influenza A virus (A/ wild duck/alberta is economized/321/88 (H9N2)) nucleoprotein (NP) gene, encoding sequence completely.

AY633295 influenza A virus (A/ wild duck/alberta is economized/11/91 (H9N2)) nucleoprotein (NP) gene, the part encoding sequence.

AY633167 influenza A virus (A/ wild duck/alberta is economized/17/91 (H9N2)) nucleoprotein (NP) gene, encoding sequence completely.

AF156410 influenza A virus (A/ quail/Hong Kong/AF157/92 (H9N2)) fragment 5 nucleoprotein (NP) gene, encoding sequence completely.

AF156414 influenza A virus (A/ quail/Arkansas State/29209-1/93 (H9N2)) fragment 5 nucleoprotein (NP) gene, the part encoding sequence.

AF156408 influenza A virus (A/ chicken/Hong Kong/739/94 (H9N2)) fragment 5 nucleoprotein (NP) gene, encoding sequence completely.

AF156409 influenza A virus (A/ chicken/Beijing/1/94 (H9N2)) fragment 5 nucleoprotein (NP) gene, encoding sequence completely.

AF536699 influenza A virus (A/ chicken/Beijing/1/95 (H9N2)) nucleoprotein (NP) gene, the part encoding sequence.

AF508596 influenza A virus (A/ ostrich/South Africa/9508103/95 (H9N2)) fragment 5 nucleoprotein (NP) gene, the part encoding sequence.

AF508600 influenza A virus (A/ duck/Germany/113/95 (H9N2)) fragment 5 nucleoprotein (NP) gene, the part encoding sequence.

AB020778 influenza A virus nucleoprotein gene, encoding sequence completely.

AF508613 influenza A virus (A/ chicken/Shandong/6/96 (H9N2)) fragment 5 nucleoprotein (NP) gene, the part encoding sequence.

AF508617 influenza A virus (A/ quail/Shanghai/8/96 (H9N2)) fragment 5 nucleoprotein (NP) gene, the part encoding sequence.

AF536703 influenza A virus (A/ chicken/Hebei/1/96 (H9N2)) nucleoprotein (NP) gene, the part encoding sequence.

AF156411 influenza A virus (A/ chicken/Korea S/38349-96323/96 (H9N2)) fragment 5 nucleoprotein (NP) gene, encoding sequence completely.

AF156412 influenza A virus (A/ chicken/Korea S/25232-96006/96 (H9N2)) fragment 5 nucleoprotein (NP) gene, the part encoding sequence.

M63779 influenza A/FPV/ the nymph of the dragonfly (Dobson)/Holland/27 (H7N7) nucleoprotein mRNA, encoding sequence completely.

M63784 j influenza A/teal (Teal)/Iceland/29/80 (H7N7) nucleoprotein mRNA, encoding sequence completely.

AJ620352 influenza A virus (A/ chicken/Germany/R28/03 (H7N7)) NP gene, geneome RNA.

AY342425 influenza A virus (A/ Holland/219/03 (H7N7)) nucleocapsid gene, encoding sequence completely.

AY342426 influenza A virus (A/ Holland/033/03 (H7N7)) nucleocapsid gene, encoding sequence completely.

AY342427 influenza A virus (A/ chicken/Holland/1/03 (H7N7)) nucleocapsid gene, encoding sequence completely.

AF156413 influenza A virus (A/ bank bird (Shorebird)/Delaware/9/96 (H9N2)) fragment 5 nucleoprotein (NP) gene, the part encoding sequence.

AF203787 influenza A virus (A/ chicken/Korea S/MS96/96 (H9N2)) nucleoprotein mRNA, encoding sequence completely.

AF156402 influenza A virus (A/ chicken/Hong Kong/G9/97 (H9N2)) fragment 5 nucleoprotein (NP) gene, encoding sequence completely.

AF156403 influenza A virus (A/ chicken/Hong Kong/G23/97 (H9N2)) fragment 5 nucleoprotein (NP) gene, encoding sequence completely.

AF156404 influenza A virus (A/ pigeon/Hong Kong/Y233/97 (H9N2)) fragment 5 nucleoprotein (NP) gene, the part encoding sequence.

AF156405 influenza A virus (A/ duck/Hong Kong/Y280/97 (H9N2)) fragment 5 nucleoprotein (NP) gene, the part encoding sequence.

AF156406 influenza A virus (A/ duck/Hong Kong/Y439/97 (H9N2)) fragment 5 nucleoprotein (NP) gene, encoding sequence completely.

AF156407 influenza A virus (A/ quail/Hong Kong/G1/97 (H9N2)) fragment 5 nucleoprotein (NP) gene, the part encoding sequence.

AF508612 influenza A virus (A/ chicken/Sichuan/5/97 (H9N2)) fragment 5 nucleoprotein (NP) gene, the part encoding sequence.

AF536702 influenza A virus (A/ chicken/Guangdong/97 (H9N2)) nucleoprotein (NP) gene, the part encoding sequence.

AF536700 influenza A virus (A/ chicken/Beijing/2/97 (H9N2)) nucleoprotein (NP) gene, the part encoding sequence.

AF508615 influenza A virus (A/ chicken/Shenzhen/9/97 (H9N2)) fragment 5 nucleoprotein (NP) gene, the part encoding sequence.

AF508616 influenza A virus (A/ duck/Nanjing/1/97 (H9N2)) fragment 5 nucleoprotein (NP) gene, the part encoding sequence.

AB049161 influenza A virus (A/ parakeet (parakeet)/Chiba (Chiba)/1/97 (H9N2)) NP gene, encoding sequence completely.

AF508603 influenza A virus (A/ pheasant/Ireland/PV18/97 (H9N2)) fragment 5 nucleoprotein (NP) gene, the part encoding sequence.

AF508607 influenza A virus (A/ chicken/Guangdong/11/97 (H9N2)) fragment 5 nucleoprotein (NP) gene, the part encoding sequence.

AF508609 influenza A virus (A/ chicken/Heilungkiang/10/97 (H9N2)) fragment 5 nucleoprotein (NP) gene, the part encoding sequence.

AF508608 influenza A virus (A/ chicken/Hebei/4/98 (H9N2)) fragment 5 nucleoprotein (NP) gene, the part encoding sequence.

AF508605 influenza A virus (A/ chicken/Beijing/8/98 (H9N2)) fragment 5 nucleoprotein (NP) gene, the part encoding sequence.

AF508599 influenza A virus (A/ chicken/Germany/R45/98 (H9N2)) fragment 5 nucleoprotein (NP) gene, the part encoding sequence.

AF536708 influenza A virus (A/ chicken/Shandong/98 (H9N2)) nucleoprotein (NP) gene, the part encoding sequence.

AY253753 influenza A virus (A/ chicken/Shanghai/F/98 (H9N2)) nucleoprotein (NP) gene, encoding sequence completely.

AF536704 influenza A virus (A/ chicken/Hebei/2/98 (H9N2)) nucleoprotein (NP) gene, the part encoding sequence.

AF536705 influenza A virus (A/ chicken/Hebei/3/98 (H9N2)) nucleoprotein (NP) gene, the part encoding sequence.

AF536706 influenza A virus (A/ chicken/Henan/98 (H9N2)) nucleoprotein (NP) gene, the part encoding sequence.

AB049162 influenza A virus (A/ parakeet/Narita Airport (Narita)/92A/98 (H9N2)) NP gene, encoding sequence completely.

AF186270 influenza A virus (A/ quail/Hong Kong/NT28/99 (H9N2)) fragment 5 nucleoprotein (NP) gene, the part encoding sequence.

AF186271 influenza A virus (A/Silkie chicken/Hong Kong/SF43/99 (H9N2)) fragment 5 nucleoprotein (NP) gene, the part encoding sequence.

AF186272 influenza A virus (A/ chicken/Hong Kong/SF2/99 (H9N2)) fragment 5 nucleoprotein (NP) gene, the part encoding sequence.

AF222614 influenza A virus (A/ quail/Hong Kong/A17/99 (H9N2)) fragment 5 nucleoprotein (NP) gene, the part encoding sequence.

AF222615 influenza A virus (A/ pigeon/Hong Kong/FY6/99 (H9N2)) fragment 5 nucleoprotein (NP) gene, the part encoding sequence.

AF222616 influenza A virus (A/ chicken/Hong Kong/NT16/99 (H9N2)) fragment 5 nucleoprotein (NP) gene, the part encoding sequence.

AF222617 influenza A virus (A/ quail/Hong Kong/SSP10/99 (H9N2)) fragment 5 nucleoprotein (NP) gene, the part encoding sequence.

AF222618 influenza A virus (A/ pheasant/Hong Kong/SSP11/99 (H9N2)) fragment 5 nucleoprotein (NP) gene, the part encoding sequence.

AF536707 influenza A virus (A/ chicken/Liaoning/99 (H9N2)) nucleoprotein (NP) gene, the part encoding sequence.

AJ291394 influenza A virus (A/ chicken/Pakistan/2/99 (H9N2)) NP gene, geneome RNA.

AF536701 influenza A virus (A/ chicken/Beijing/3/99 (H9N2)) nucleoprotein (NP) gene, the part encoding sequence.

AF508611 influenza A virus (A/ chicken/Ningxia/5/99 (H9N2)) fragment 5 nucleoprotein (NP) gene, the part encoding sequence.

AF508614 influenza A virus (A/ chicken/Shijiazhuang/2/99 (H9N2)) fragment 5 nucleoprotein (NP) gene, the part encoding sequence.

AF508604 influenza A virus (A/ chicken/Korea S/99029/99 (H9N2)) fragment 5 nucleoprotein (NP) gene, the part encoding sequence.

AF222619 influenza A virus (A/ chicken/Hong Kong/FY20/99 (H9N2)) fragment 5 nucleoprotein (NP) gene, the part encoding sequence.

AF222620 influenza A virus (A/ chicken/Hong Kong/KC12/99 (H9N2)) fragment 5 nucleoprotein (NP) gene, the part encoding sequence.

AF222621 influenza A virus (A/ Gallus Domesticus/Hong Kong/SF44/99 (H9N2)) fragment 5 nucleoprotein (NP) gene, the part encoding sequence.

AF508601 influenza A virus (A/ chicken/Iran/11T/99 (H9N2)) fragment 5 nucleoprotein (NP) gene, the part encoding sequence.

AF508602 influenza A virus (A/ chicken/Saudi Arabia/532/99 (H9N2)) fragment 5 nucleoprotein (NP) gene, the part encoding sequence.

AF508597 influenza A virus (A/ chicken/Pakistan/4/99 (H9N2)) fragment 5 nucleoprotein (NP) gene, the part encoding sequence.

AF508598 influenza A virus (A/ chicken/Pakistan/5/99 (H9N2)) fragment 5 nucleoprotein (NP) gene, the part encoding sequence.

AF508606 influenza A virus (A/ chicken/Guangdong/10/00 (H9N2)) fragment 5 nucleoprotein (NP) gene, the part encoding sequence.

AF508610 influenza A virus (A/ chicken/Henan/62/00 (H9N2)) fragment 5 nucleoprotein (NP) gene, the part encoding sequence.

AF523410 influenza A virus (A/ duck/Shantou/1043/00 (H9N2)) nucleocapsid protein (NP) gene, the part encoding sequence.

AF523411 influenza A virus (A/ duck/Shantou/2134/00 (H9N2)) nucleocapsid protein (NP) gene, the part encoding sequence.

AF523413 influenza A virus (A/ duck/Shantou/1042/00 (H9N2)) nucleocapsid protein (NP) gene, the part encoding sequence.

AF523415 influenza A virus (A/ duck/Shantou/2102/00 (H9N2)) nucleocapsid protein (NP) gene, the part encoding sequence.

AF523416 influenza A virus (A/ duck/Shantou/830/00 (H9N2)) nucleocapsid protein (NP) gene, the part encoding sequence.

AF523417 influenza A virus (A/ duck/Shantou/2144/00 (H9N2)) nucleocapsid protein (NP) gene, the part encoding sequence.

AF523419 influenza A virus (A/ duck/Shantou/2143/00 (H9N2)) nucleocapsid protein (NP) gene, the part encoding sequence.

AF523420 influenza A virus (A/ duck/Shantou/1881/00 (H9N2)) nucleocapsid protein (NP) gene, the part encoding sequence.

AJ427864 influenza A virus (A/ quail/Hong Kong/FY298/00 (H9N2)) part nucleoprotein np gene, geneome RNA

AY180525 influenza A virus (A/ pigeon/Nanchang/2-0461/2000 (H9N2)) nucleoprotein (NP) gene, the part encoding sequence.

AY180534 influenza A virus strain A/ duck/Nanchang/7-092/2000 (H9N2) nucleoprotein (NP) gene, the part encoding sequence.

AY180537 influenza A virus strain A/ duck/Nanchang/11-392/2000 (H9N2) nucleoprotein (NP) gene, the part encoding sequence.

AY180538 influenza A virus (A/ pigeon/Nanchang/11-145/2000 (H9N2)) nucleoprotein (NP) gene, the part encoding sequence.

AY180542 influenza A virus strain A/ duck/Nanchang/11-197/2000 (H9N2) nucleoprotein (NP) gene, the part encoding sequence.

AY180544 influenza A virus strain A/ duck/Nanchang/11-290/2000 (H9N2) nucleoprotein (NP) gene, the part encoding sequence.

AY180560 influenza A virus (A/ pigeon/Nanchang/7-058/2000 (H9N2)) nucleoprotein (NP) gene, the part encoding sequence.

AY180562 influenza A virus strain A/ chicken/Nanchang/4-010/2000 (H9N2) nucleoprotein (NP) gene, the part encoding sequence.

AY180563 influenza A virus strain A/ quail/Nanchang/4-040/2000 (H9N2) nucleoprotein (NP) gene, the part encoding sequence.

AY180564 influenza A virus (A/ wild duck/Nanchang/2-0480/2000 (H9N2)) nucleoprotein (NP) gene, the part encoding sequence.

AY180575 influenza A virus (A/ quail/Nanchang/2-0460/2000 (H9N2)) nucleoprotein (NP) gene, the part encoding sequence.

AY496851 influenza A virus (A/ chicken/Mudanjiang/0823/2000 (H9N2)) nucleoprotein (np) mRNA, encoding sequence completely.

AY180581 influenza A virus (A/ chicken/Nanchang/1-0016/2000 (H9N2)) nucleoprotein (NP) gene, the part encoding sequence.

AY180583 influenza A virus strain A/ duck/Nanchang/10-389/2000 (H9N2) nucleoprotein (NP) gene, the part encoding sequence.

AY180584 influenza A virus strain A/ duck/Nanchang/1-0070/2000 (H9N2) nucleoprotein (NP) gene, the part encoding sequence.

AY768567 influenza A virus (A/ chicken/Korea S/SNU0028/00 (H9N2)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY768568 influenza A virus (A/ chicken/Korea S/SNU0037/00 (H9N2)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY768569 influenza A virus (A/ chicken/Korea S/SNU0057/00 (H9N2)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY768570 influenza A virus (A/ chicken/Korea S/SNU0073/00 (H9N2)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY768571 influenza A virus (A/ chicken/Korea S/SNU0091/00 (H9N2)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY768572 influenza A virus (A/ chicken/Korea S/SNU0140/00 (H9N2)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY768573 influenza A virus (A/ chicken/Korea S/SNU0146/00 (H9N2)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY768574 influenza A virus (A/ chicken/Korea S/SNU1035C/00 (H9N2)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY268949 influenza A virus (A/ chicken/Wangcheng (Wangcheng)/4/2001 (H9N2)) nucleoprotein mRNA, encoding sequence completely.

AY180578 influenza A virus strain A/ chicken/Nanchang/4-301/2001 (H9N2) nucleoprotein (NP) gene, the part encoding sequence.

AY180551 influenza A virus (A/ chicken/Nanchang/4-361/2001 (H9N2)) nucleoprotein (NP) gene, the part encoding sequence.

AF523418 influenza A virus (A/ duck/Shantou/2088/01 (H9N2)) nucleocapsid protein (NP) gene, the part encoding sequence.

AF523414 influenza A virus (A/ duck/Shantou/1605/01 (H9N2)) nucleocapsid protein (NP) gene, the part encoding sequence.

AF523412 influenza A virus (A/Wild duck/Shantou/4808/01 (H9N2)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY800236 influenza A virus (A/ chicken/Korea S/S1/2003 (H9N2)) nucleoprotein (NP) gene, the part encoding sequence.

AY862646 influenza A virus (A/silky chicken/Korea S/S3/03 (H9N2)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY862647 influenza A virus (A/ chicken/Korea S/S4/03 (H9N2)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY862648 influenza A virus (A/ chicken/Korea S/S5/03 (H9N2)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY862649 influenza A virus (A/ chicken/Korea S/S12/03 (H9N2)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY862650 influenza A virus (A/ duck/Korea S/S13/03 (H9N2)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY862651 influenza A virus (A/ dove/Korea S/S14/03 (H9N2)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY862652 influenza A virus (A/ chicken/Korea S/S15/03 (H9N2)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY862653 influenza A virus (A/ chicken/Korea S/S16/03 (H9N2)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY862654 influenza A virus (A/ chicken/Korea S/S18/03 (H9N2)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY664717 influenza A virus (A/ chicken/Hong Kong/CSW153/03 (H9N2)) nucleoprotein (NP) gene, encoding sequence completely.

AY664718 influenza A virus (A/ chicken/Hong Kong/AP45/03 (H9N2)) nucleoprotein (NP) gene, encoding sequence completely.

AY664719 influenza A virus (A/ chicken/Hong Kong/BD90/03 (H9N2)) nucleoprotein (NP) gene, encoding sequence completely.

AY664720 influenza A virus (A/ chicken/Hong Kong/CSW291/03 (H9N2)) nucleoprotein (NP) gene, encoding sequence completely.

AY664721 influenza A virus (A/ chicken/Hong Kong/CSW304/03 (H9N2)) nucleoprotein (NP) gene, encoding sequence completely.

AY664722 influenza A virus (A/ chicken/Hong Kong/FY23/03 (H9N2)) nucleoprotein (NP) gene, encoding sequence completely.

AY664723 influenza A virus (A/ vulture galeeny/Hong Kong/NT101/03 (H9N2)) nucleoprotein (NP) gene, encoding sequence completely.

AY664724 influenza A virus (A/ chicken/Hong Kong/NT142/03 (H9N2)) nucleoprotein (NP) gene, encoding sequence completely.

AY664725 influenza A virus (A/ chicken/Hong Kong/SF1/03 (H9N2)) nucleoprotein (NP) gene, encoding sequence completely.

AY664726 influenza A virus (A/ chicken/Hong Kong/SSP101/03 (H9N2)) nucleoprotein (NP) gene, encoding sequence completely.

AY664727 influenza A virus (A/ chicken/Hong Kong/TP38/03 (H9N2)) nucleoprotein (NP) gene, encoding sequence completely.

AY664728 influenza A virus (A/ chicken/Hong Kong/WF126/03 (H9N2)) nucleoprotein (NP) gene, encoding sequence completely.

AY664729 influenza A virus (A/ pigeon/Hong Kong/WF53/03 (H9N2)) nucleoprotein (NP) gene, encoding sequence completely.

AY664730 influenza A virus (A/ pheasant/Hong Kong/WF54/03 (H9N2)) nucleoprotein (NP) gene, encoding sequence completely.

AY664731 influenza A virus (A/ vulture galeeny/Hong Kong/NT184/03 (H9N2)) nucleoprotein (NP) gene, encoding sequence completely.

AY664732 influenza A virus (A/ chicken/Hong Kong/WF120/03 (H9N2)) nucleoprotein (NP) gene, encoding sequence completely.

AY664733 influenza A virus (A/ chicken/Hong Kong/NT366/03 (H9N2)) nucleoprotein (NP) gene, the part encoding sequence.

AY664734 influenza A virus (A/ chicken/Hong Kong/SSP418/03 (H9N2)) nucleoprotein (NP) gene, the part encoding sequence.

AY664735 influenza A virus (A/ chicken/Hong Kong/YU427/03 (H9N2)) nucleoprotein (NP) gene, the part encoding sequence.

AY788915 influenza A virus (A/ chicken/China/HSS2004 (H9N2)) nucleoprotein (NP) gene, encoding sequence completely.

AY586423 influenza A virus (A/ wild duck/Italy/33/01 (H7N3)) nucleoprotein gene, the part encoding sequence.

AY586424 influenza A virus (A/ wild duck/Italy/43/01 (H7N3)) nucleoprotein gene, the part encoding sequence.

AY586425 influenza A virus (A/ turkey/Italy/220158/2002 (H7N3)) nucleoprotein gene, the part encoding sequence.

AY586426 influenza A virus (A/ turkey/Italy/214845/02 (H7N3)) nucleoprotein gene, the part encoding sequence.

AJ627486 influenza A virus (A/ turkey/Italy/214845/2002 (H7N3)) NP gene, geneome RNA.

AJ627495 influenza A virus (A/ turkey/Italy/220158/2002 (H7N3)) NP gene, geneome RNA.

AY303658 influenza A virus (A/ chicken/Chile/176822/02 (H7N3)) nucleoprotein gene, encoding sequence completely.

AY303659 influenza A virus (A/ chicken/Chile/4957/02 (H7N3)) nucleoprotein gene, encoding sequence completely.

AY611527 influenza A virus (A/ chicken/British Columbia/04 (H7N3)) nucleoprotein (NP) gene, encoding sequence completely.

AY646081 influenza A virus (A/ chicken/British Columbia/GSC_ people _ B/04 (H7N3)) nucleoprotein (NP) gene, encoding sequence completely.

AY648290 influenza A virus (A/GSC_ chicken _ B/ British Columbia/04 (H7N3)) nucleoprotein (NP) gene, encoding sequence completely.

AY650273 influenza A virus (A/GSC_ chicken/British Columbia/04 (H7N3)) nucleoprotein (NP) gene, encoding sequence completely.

AF144303 influenza A virus (A/ goose/Guangdong/1/96 (H5N1)) nucleocapsid protein (NP) gene, encoding sequence completely.

AF046084 influenza A virus (A/ chicken/Hong Kong/220/97 (H5N1)) nucleoprotein gene, encoding sequence completely.

AF057293 influenza A virus (A/ chicken/Hong Kong/258/97 (H5N1)) nucleoprotein mRNA, encoding sequence completely.

AF098617 influenza A virus (A/ chicken/Hong Kong/y388/97 (H5N1)) nucleoprotein (NP) gene, encoding sequence completely.

AF098618 influenza A virus (A/ chicken/Hong Kong/728/97 (H5N1)) nucleoprotein (NP) gene, encoding sequence completely.

AF098619 influenza A virus (A/ chicken/Hong Kong/786/97 (H5N1)) nucleoprotein (NP) gene, encoding sequence completely.

AF098620 influenza A virus (A/ chicken/Hong Kong/915/97 (H5N1)) nucleoprotein (NP) gene, encoding sequence completely.

AF098621 influenza A virus (A/ duck/Hong Kong/p46/97 (H5N1)) nucleoprotein (NP) gene, encoding sequence completely.

AF098622 influenza A virus (A/ duck/Hong Kong/y283/97 (H5N1)) nucleoprotein (NP) gene, encoding sequence completely.

AF098623 influenza A virus (A/ goose/Hong Kong/w355/97 (H5N1)) nucleoprotein (NP) gene, encoding sequence completely.

AF370122 influenza A virus (A/ goose/Guangdong/3/97 (H5N1)) nucleoprotein gene, encoding sequence completely.

AF216712 influenza A virus (A/ environment/Hong Kong/437-4/99 (H5N1)) nucleoprotein gene, encoding sequence completely.

AF216720 influenza A virus (A/ environment/Hong Kong/437-6/99 (H5N1)) nucleoprotein gene, encoding sequence completely.

AF216728 influenza A virus (A/ environment/Hong Kong/437-8/99 (H5N1)) nucleoprotein gene, encoding sequence completely.

AF216736 influenza A virus (A/ environment/Hong Kong/437-10/99 (H5N1)) nucleoprotein gene, encoding sequence completely.

AY585429 influenza A virus (A/ duck/Guangxi/07/1999 (H5N1)) nucleoprotein (NP) mRNA, encoding sequence completely.

AY585439 influenza A virus (A/ duck/Zhejiang/11/2000 (H5N1)) nucleoprotein (NP) mRNA, encoding sequence completely.

AY585440 influenza A virus (A/ duck/Zhejiang/52/2000 (H5N1)) nucleoprotein (NP) mRNA, encoding sequence completely.

AY585428 influenza A virus (A/ duck/Guangdong/40/2000 (H5N1)) nucleoprotein (NP) mRNA, encoding sequence completely.

AY585423 influenza A virus (A/ duck/Fujian/19/2000 (H5N1)) nucleoprotein (NP) mRNA, encoding sequence completely.

AY585425 influenza A virus (A/ duck/Guangdong/07/2000 (H5N1)) nucleoprotein (NP) mRNA, encoding sequence completely.

AY585426 influenza A virus (A/ duck/Guangdong/12/2000 (H5N1)) nucleoprotein (NP) mRNA, encoding sequence completely.

AY059492 influenza A virus (A/ goose/Hong Kong/ww26/2000 (H5N1)) fragment 5 nucleocapsid proteins (NP) gene, the part encoding sequence.

AY059493 influenza A virus (A/ goose/Hong Kong/ww28/2000 (H5N1)) fragment 5 nucleocapsid proteins (NP) gene, the part encoding sequence.

AY059494 influenza A virus (A/ duck/Hong Kong/ww381/2000 (H5N1)) fragment 5 nucleocapsid proteins (NP) gene, the part encoding sequence.

AY059495 influenza A virus (A/ duck/Hong Kong/ww461/2000 (H5N1)) fragment 5 nucleocapsid proteins (NP) gene, the part encoding sequence.

AY059496 influenza A virus (A/ goose/Hong Kong/ww491/2000 (H5N1)) fragment 5 nucleocapsid proteins (NP) gene, the part encoding sequence.

AY059497 influenza A virus (A/ duck/Hong Kong/2986.1/2000 (H5N1)) fragment 5 nucleocapsid proteins (NP) gene, the part encoding sequence.

AY059498 influenza A virus (A/ goose/Hong Kong/3014.8/2000 (H5N1)) fragment 5 nucleocapsid proteins (NP) gene, the part encoding sequence.

AF398419 influenza A virus (A/ goose/Hong Kong/385.3/2000 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AF398420 influenza A virus (A/ goose/Hong Kong/385.5/2000 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AF468842 influenza A virus (A/ duck/Anyang/AVL-1/2001 (H5N1)) nucleoprotein (NP) gene, encoding sequence completely.

AF509117 influenza A virus (A/ chicken/Hong Kong/FY77/01 (H5N1)) nucleocapsid protein (NP) gene, encoding sequence completely.

AF509118 influenza A virus (A/ chicken/Hong Kong/YU562/01 (H5N1)) nucleocapsid protein (NP) gene, encoding sequence completely.

AF509119 influenza A virus (A/ chicken/Hong Kong/YU563/01 (H5N1)) nucleocapsid protein (NP) gene, encoding sequence completely.

AY585438 influenza A virus (A/ duck/Shanghai/38/2001 (H5N1)) nucleoprotein (NP) mRNA, encoding sequence completely.

AY221548 influenza A virus (A/ chicken/Hong Kong/NT873.3/01-MB (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY221549 influenza A virus (A/ chicken/Hong Kong/NT873.3/01 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY221550 influenza A virus (A/ chicken/Hong Kong/FY150/01-MB (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY221551 influenza A virus (A/ chicken/Hong Kong/FY150/01 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY221552 influenza A virus (A/ pheasant/Hong Kong/FY155/01-MB (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY221553 influenza A virus (A/ pheasant/Hong Kong/FY155/01 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY221554 influenza A virus (A/ chicken/Hong Kong/YU822.2/01-MB (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY221555 influenza A virus (A/ chicken/Hong Kong/YU822.2/01 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY221556 influenza A virus (A/ chicken/Hong Kong/YU562/01 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AF509120 influenza A virus (A/ chicken/Hong Kong/FY150/01 (H5N1)) nucleocapsid protein (NP) gene, encoding sequence completely.

AF509121 influenza A virus (A/ pheasant/Hong Kong/FY155/01 (H5N1)) nucleocapsid protein (NP) gene, encoding sequence completely.

AF509122 influenza A virus (A/ Gallus Domesticus/Hong Kong/SF189/01 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AF509123 influenza A virus (A/ quail/Hong Kong/SF203/01 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AF509124 influenza A virus (A/ pigeon/Hong Kong/SF215/01 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AF509125 influenza A virus (A/ chicken/Hong Kong/SF219/01 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AF509126 influenza A virus (A/ chicken/Hong Kong/715.5/01 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AF509127 influenza A virus (A/ chicken/Hong Kong/751.1/01 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AF509128 influenza A virus (A/ chicken/Hong Kong/822.1/01 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AF509129 influenza A virus (A/ chicken/Hong Kong/829.2/01 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AF509130 influenza A virus (A/ chicken/Hong Kong/830.2/01 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AF509131 influenza A virus (A/ chicken/Hong Kong/858.3/01 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AF509132 influenza A virus (A/ chicken/Hong Kong/866.3/01 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AF509133 influenza A virus (A/ chicken/Hong Kong/867.1/01 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AF509134 influenza A virus (A/ chicken/Hong Kong/879.1/01 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AF509135 influenza A virus (A/ chicken/Hong Kong/873.3/01 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AF509136 influenza A virus (A/ chicken/Hong Kong/876.1/01 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AF509137 influenza A virus (A/ chicken/Hong Kong/891.1/01 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AF509138 influenza A virus (A/ chicken/Hong Kong/893.2/01 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AF509139 influenza A virus (A/ goose/Hong Kong/76.1/01 (H5N1)) nucleocapsid protein (NP) gene, encoding sequence completely.

AF509140 influenza A virus (A/ goose/Hong Kong/ww100/01 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AF509141 influenza A virus (A/ duck/Hong Kong/573.4/01 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AF509142 influenza A virus (A/ duck/Hong Kong/646.3/01 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY585424 influenza A virus (A/ duck/Guangdong/01/2001 (H5N1)) nucleoprotein (NP) mRNA, encoding sequence completely.

AY585422 influenza A virus (A/ duck/Fujian/17/2001 (H5N1)) nucleoprotein (NP) mRNA, encoding sequence completely.

AY585430 influenza A virus (A/ duck/Guangxi/22/2001 (H5N1)) nucleoprotein (NP) mRNA, encoding sequence completely.

AY585431 influenza A virus (A/ duck/Guangxi/35/2001 (H5N1)) nucleoprotein (NP) mRNA, encoding sequence completely.

AY585432 influenza A virus (A/ duck/Guangxi/50/2001 (H5N1)) nucleoprotein (NP) mRNA, encoding sequence completely.

AY585434 influenza A virus (A/ duck/Shanghai/08/2001 (H5N1)) nucleoprotein (NP) mRNA, encoding sequence completely.

AY585435 influenza A virus (A/ duck/Shanghai/13/2001 (H5N1)) nucleoprotein (NP) mRNA, encoding sequence completely.

AY585436 influenza A virus (A/ duck/Shanghai/35/2002 (H5N1)) nucleoprotein (NP) mRNA, encoding sequence completely.

AY585437 influenza A virus (A/ duck/Shanghai/37/2002 (H5N1)) nucleoprotein (NP) mRNA, encoding sequence completely.

AY585433 influenza A virus (A/ duck/Guangxi/53/2002 (H5N1)) nucleoprotein (NP) mRNA, encoding sequence completely.

AY585427 influenza A virus (A/ duck/Guangdong/22/2002 (H5N1)) nucleoprotein (NP) mRNA, encoding sequence completely.

AY585420 influenza A virus (A/ duck/Fujian/01/2002 (H5N1)) nucleoprotein (NP) mRNA, encoding sequence completely.

AY585421 influenza A virus (A/ duck/Fujian/13/2002 (H5N1)) nucleoprotein (NP) mRNA, encoding sequence completely.

AY575907 influenza A virus (A/Gs/ Hong Kong/739.2/02 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY575908 influenza A virus (A/Eg/ Hong Kong/757.3/02 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY575909 influenza A virus (A/G.H/ Hong Kong/793.1/02 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY575910 influenza A virus (A/Dk/ Hong Kong/821/02 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY575911 influenza A virus (A/Ck/ Hong Kong/31.4/02 (H5N1)) nucleocapsid protein (NP) gene, encoding sequence completely.

AY575912 influenza A virus (A/Ck/ Hong Kong/61.9/02 (H5N1)) nucleocapsid protein (NP) gene, encoding sequence completely.

AY575913 influenza A virus (A/Ck/ Hong Kong/YU777/02 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY575914 influenza A virus (A/Ck/ Hong Kong/96.1/02 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY575915 influenza A virus (A/Ck/ Hong Kong/409.1/02 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY575916 influenza A virus (A/Ph/ Hong Kong/sv674.15/02 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

DQ023146 influenza A virus (A/ chicken/sd/1/02 (H5N1)) nucleoprotein (NP) mRNA, encoding sequence completely.

AY676037 influenza A virus (A/ duck/Hong Kong/821/02 (H5N1)) nucleoprotein (NP) gene, encoding sequence completely.

AY651510 influenza A virus (A/Gf/ Hong Kong/38/2002 (H5N1)) nucleocapsid protein (NP) gene, encoding sequence completely.

AY651511 influenza A virus (A/Ck/ Hong Kong/31.2/2002 (H5N1)) nucleocapsid protein (NP) gene, encoding sequence completely.

AY651512 influenza A virus (A/Ck/ Hong Kong/37.4/2002 (H5N1)) nucleocapsid protein (NP) gene, encoding sequence completely.

AY651513 influenza A virus (A/SCk/ Hong Kong/YU100/2002 (H5N1)) nucleocapsid protein (NP) gene, encoding sequence completely.

AY651514 influenza A virus (A/Ck/ Hong Kong/YU22/2002 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY651521 influenza A virus (A/Ck/ Hong Kong/3176.3/2002 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY651522 influenza A virus (A/Ck/ Hong Kong/3169.1/2002 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY651524 influenza A virus (A/ rock dove/Hong Kong/862.7/2002 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY651525 influenza A virus (A/ sets sparrow/Hong Kong/864/2002 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY651526 influenza A virus (A/ heron/Hong Kong/861.1/2002 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY651527 influenza A virus (A/ teal/China/2978.1/2002 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY651523 influenza A virus (the red mouth of A/ gull/Hong Kong/12.1/2003 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY651487 influenza A virus (A/Ck/ Indonesia/PA/2003 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY651515 influenza A virus (A/Ck/ Hong Kong/2133.1/2003 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY651516 influenza A virus (A/Ck/ Hong Kong/NT93/2003 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY651517 influenza A virus (A/Ck/ Hong Kong/SSP141/2003 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY651518 influenza A virus (A/Ck/ Hong Kong/WF157/2003 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY651519 influenza A virus (A/Ck/ Hong Kong/FY157/2003 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY651520 influenza A virus (A/Ck/ Hong Kong/YU324/2003 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY651490 influenza A virus (A/Ck/ Indonesia/2A/2003 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY676038 influenza A virus (A/ egression (egret)/Hong Kong/757.2/03 (H5N1)) nucleoprotein (NP) gene, encoding sequence completely.

AY676039 influenza A virus (A/ chicken/Korea S/ES/03 (H5N1)) nucleoprotein (NP) gene, encoding sequence completely.

AY676040 influenza A virus (A/ duck/Korea S/ESD1/03 (H5N1)) nucleoprotein (NP) gene, encoding sequence completely.

AY651529 influenza A virus (A/Dk/HN/5806/2003 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY651532 influenza A virus (A/Ck/ST/4231/2003 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY651534 influenza A virus (A/Dk/ST/4003/2003 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY651535 influenza A virus (A/Dk/YN/6255/2003 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY651536 influenza A virus (A/Dk/YN/6445/2003 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY651485 influenza A virus (A/Ck/ Indonesia/BL/2003 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY518364 influenza A virus (A/ duck/China/E319-2/03 (H5N1)) nucleocapsid protein (NP) gene, encoding sequence completely.

AY574189 influenza A virus (A/ chicken/Vietnam/HD1/2004 (H5N1)) nucleoprotein gene, the part encoding sequence.

AY574192 influenza A virus (A/ chicken/Vietnam/HD2/2004 (H5N1)) nucleoprotein gene, the part encoding sequence.

AJ867076 influenza A virus (A/Hatay/2004/ (H5N1)) NP gene, geneome RNA

AY651486 influenza A virus (A/Dk/ Indonesia/MS/2004 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY590579 influenza A virus (A/ chicken/Buddhist system/Thailand/CU-K2/2004 (H5N1)) nucleocapsid protein (NP) gene, encoding sequence completely.

AY609313 influenza A virus (A/ chicken/Guangdong/174/04 (H5N1)) fragment 5, sequence completely.

AY576929 influenza A virus (A/ chicken/Vietnam/CM/2004 (H5N1)) fragment 5 nucleoprotein genes, the part encoding sequence.

AY576931 influenza A virus (A/ kind of duck (muscovy duck)/Vietnam/MdGL/2004 (H5N1)) fragment 5 nucleoprotein genes, the part encoding sequence.

AB166863 influenza A virus (A/ chicken/mountain pass (Yamaguchi)/7/2004 (H5N1)) NP gene, encoding sequence completely.

AB188817 influenza A virus (A/ chicken/big/8/2004 (H5N1) that divide) NP gene, encoding sequence completely.

AY651537 influenza A virus (A/Ck/YN/374/2004 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY651538 influenza A virus (A/Ck/YN/115/2004 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY653196 influenza A virus (A/ chicken/Jilin/9/2004 (H5N1)) fragment 5, sequence completely.

AY651533 influenza A virus (A/Ph/ST/44/2004 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY651530 influenza A virus (A/Dk/HN/303/2004 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY651531 influenza A virus (A/Dk/HN/101/2004 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY684707 influenza A virus (A/ chicken/Hubei/327/2004 (H5N1)) nucleoprotein (NP) gene, encoding sequence completely.

AY737290 influenza A virus (A/ chicken/Guangdong/191/04 (H5N1)) fragment 5, sequence completely.

AY737297 influenza A virus (A/ chicken/Guangdong/178/04 (H5N1)) fragment 5, sequence completely.

AY737305 influenza A virus (A/ duck/Guangdong/173/04 (H5N1)) fragment 5, sequence completely.

AY770081 influenza A virus (A/ chicken/Hubei/489/2004 (H5N1)) nucleoprotein (NP) gene, encoding sequence completely.

AY770996 influenza A virus (A/ chicken/big subtlety/Thailand/CU-23/04 (H5N1)) nucleoprotein gene, the part encoding sequence.

AY818139 influenza A virus (A/ chicken/Vietnam/C58/04 (H5N1)) NP gene, encoding sequence completely.

AY818140 influenza A virus (A/ quail/Vietnam/36/04 (H5N1)) NP gene, encoding sequence completely.

AY856864 influenza A virus (A/ duck/Shandong/093/2004 (H5N1)) fragment 5, sequence completely.

AB189046 influenza A virus (A/ chicken/capital of a country/3/2004 (H5N1)) NP gene, encoding sequence completely.

AB189054 influenza A virus (A/ crow/capital of a country/53/2004 (H5N1)) NP gene, encoding sequence completely.

AB189062 influenza A virus (A/ crow/Osaka/102/2004 (H5N1)) NP gene, encoding sequence completely.

AY651491 influenza A virus (A/Ck/ Thailand/1/2004 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY651492 influenza A virus (A/Ck/ Thailand/73/2004 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY651493 influenza A virus (A/Ck/ Thailand/9.1/2004 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY651494 influenza A virus (A/Qa/ Thailand/57/2004 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY651495 influenza A virus (A/bird/ Thailand/3.1/2004 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY651496 influenza A virus (A/Dk/ Thailand/71.1/2004 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY651497 influenza A virus (A/Gs/ Thailand/79/2004 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY651502 influenza A virus (A/Ck/ Vietnam/33/2004 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY651503 influenza A virus (A/Ck/ Vietnam/35/2004 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY651504 influenza A virus (A/Ck/ Vietnam/36/2004 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY651505 influenza A virus (A/Ck/ Vietnam/37/2004 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY651506 influenza A virus (A/Ck/ Vietnam/38/2004 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY651507 influenza A virus (A/Ck/ Vietnam/39/2004 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY651508 influenza A virus (A/Ck/ Vietnam/C57/2004 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY651509 influenza A virus (A/Dk/ Vietnam/11/2004 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY651488 influenza A virus (A/Ck/ Indonesia/4/2004 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY651489 influenza A virus (A/Ck/ Indonesia/5/2004 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY651528 influenza A virus (A/ falcon/Hong Kong/D0028/2004 (H5N1)) nucleocapsid protein (NP) gene, the part encoding sequence.

M22573 influenza A/duck/Hong Kong/7/75 (H3N2), nucleoprotein (fragment (seg) 5), RNA.

AY180555 influenza A virus (A/ chicken/Nanchang/3-120/2001 (H3N2)) nucleoprotein (NP) gene, the part encoding sequence.

AY779261 influenza A virus (A/ turkey/North Carolina state/12344/03 (H3N2)) nucleoprotein (NP) gene, the part encoding sequence.

AY779262 influenza A virus (A/ turkey/Minnesota State/764-2/03 (H3N2)) nucleoprotein (NP) gene, the part encoding sequence.

AY862655 influenza A virus (A/ chicken/Korea S/S6/03 (H3N2)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY862656 influenza A virus (A/ duck/Korea S/S7/03 (H3N2)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY862657 influenza A virus (A/ duck/Korea S/S8/03 (H3N2)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY862658 influenza A virus (A/ duck/Korea S/S9/03 (H3N2)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY862659 influenza A virus (A/ duck/Korea S/S10/03 (H3N2)) nucleocapsid protein (NP) gene, the part encoding sequence.

AY862660 influenza A virus (A/ dove/Korea S/S11/03 (H3N2)) nucleocapsid protein (NP) gene, the part encoding sequence.

D00050 influenza A virus nucleoprotein gene, encoding sequence completely.

M14921 influenza A/wild duck/NY/6750/78 (H2N2) nucleoprotein (fragment 5) RNA, encoding sequence completely.

AY422026 influenza A virus (A/ duck/Hokkaido/95/01 (H2N2)) nucleoprotein (NP) gene, the part encoding sequence.

U49093 influenza A virus nucleoprotein (NP) mRNA, the part encoding sequence.

M22574 influenza A/duck/Bavaria/2/77 (H1N1), nucleoprotein (fragment 5), RNA.

M76603 influenza A/turkey/England/647/77 (H1N1) mRNA, encoding sequence completely.

M63783 influenza A/duck/Australia/749/80 (H1N1) nucleoprotein mRNA, encoding sequence completely.

M63778 influenza A/turkey/the Minnesota State/1661/81 (H1N1) nucleoprotein mRNA, encoding sequence completely.

Z26855 influenza A virus NP gene

M76609 influenza A/turkey/the North Carolina state/1790/88 (H1N1) mRNA, encoding sequence completely.

Z26857 influenza A virus NP gene

AF213905 influenza A virus (A/ wild duck/Italy/24/95 (H1N1)) fragment 5 nucleoprotein (NP) gene, the part encoding sequence.

AF213906 influenza A virus (A/ chicken/Italy/24/95 (H1N1)) fragment 5 nucleoprotein (NP) gene, the part encoding sequence.

AY633215 influenza A virus (A/ wild duck/alberta is economized/211/98 (H1N1)) nucleoprotein (NP) gene, encoding sequence completely.

AY180543 influenza A virus (A/ quail/Nanchang/12-340/2000 (H1N1)) nucleoprotein (NP) gene, the part encoding sequence.

Used sequence during influenza A polysaccharase basic protein 1 (PB1) is analyzed

The P1 of RNA polymerase subunit (PB1) gene that AY633218 influenza A virus (A/ wild duck/alberta is economized/211/98 (H1N1)) RNA instructs, the part encoding sequence.

U48284 influenza A virus polysaccharase (PB1) mRNA, the part encoding sequence.

AY180855 influenza A virus strain A/ quail/Nanchang/12-340/2000 (H1N1) the polysaccharase PB1 of subunit (PB1) gene, the part encoding sequence.

AY422038 influenza A virus (A/ duck/Hokkaido/95/01 (H2N2)) polysaccharase subunit (PB1) gene, the part encoding sequence.

M25926 influenza A/wild duck/New York/6750/78 (H2N2) PB1 gene, encoding sequence completely.

AY180871 influenza A virus strain A/ chicken/Nanchang/3-120/2001 (H3N2) the polysaccharase PB1 of subunit (PB1) gene, the part encoding sequence.

AY779265 influenza A virus (A/ turkey/North Carolina state/12344/03 (H3N2)) polysaccharase basic protein 1 (PB1) gene, the part encoding sequence.

AY779266 influenza A virus (A/ turkey/Minnesota State/764-2/03 (H3N2)) polysaccharase basic protein 1 (PB1) gene, the part encoding sequence.

AY862703 influenza A virus (A/ chicken/Korea S/S6/03 (H3N2)) PB1 (PB1) gene, the part encoding sequence.

AY862704 influenza A virus (A/ duck/Korea S/S7/03 (H3N2)) PB1 (PB1) gene, the part encoding sequence.

AY862705 influenza A virus (A/ duck/Korea S/S8/03 (H3N2)) PB1 (PB1) gene, the part encoding sequence.

AY862706 influenza A virus (A/ duck/Korea S/S9/03 (H3N2)) PB1 (PB1) gene, the part encoding sequence.

AY862707 influenza A virus (A/ duck/Korea S/S10/03 (H3N2)) PB1 (PB1) gene, the part encoding sequence.

AY862708 influenza A virus (A/ dove/Korea S/S11/03 (H3N2)) PB1 (PB1) gene, the part encoding sequence.

AF213911 influenza A virus (A/ chicken/Italy/5945/95 (H3N2)) fragment 8PB1 polymerase protein gene, the part encoding sequence.

AB166860 influenza A virus (A/ chicken/mountain pass/7/2004 (H5N1)) polysaccharase basic protein 1 (PB1) gene, encoding sequence completely.

AB188814 influenza A virus (A/ chicken/big/8/2004 (H5N1) that divide) polysaccharase basic protein 1 (PB1) gene, encoding sequence completely.

AF398423 influenza A virus (A/ goose/Hong Kong/385.3/2000 (H5N1)) polysaccharase (PB1) gene, the part encoding sequence.

AF398424 influenza A virus (A/ goose/Hong Kong/385.5/2000 (H5N1)) polysaccharase (PB1) gene, the part encoding sequence.

AF468839 influenza A virus (A/ duck/Anyang/AVL-1/2001 (H5N1)) polysaccharase basic protein 1 (PB1) gene, encoding sequence completely.

AF509169 influenza A virus (A/ chicken/Hong Kong/FY77/01 (H5N1)) polysaccharase (PB1) gene, the part encoding sequence.

AF509170 influenza A virus (A/ chicken/Hong Kong/YU562/01 (H5N1)) polysaccharase (PB1) gene, the part encoding sequence.

AF509171 influenza A virus (A/ chicken/Hong Kong/YU563/01 (H5N1)) polysaccharase (PB1) gene, the part encoding sequence.

AF509172 influenza A virus (A/ chicken/Hong Kong/FY150/01 (H5N1)) polysaccharase (PB1) gene, the part encoding sequence.

AF509173 influenza A virus (A/ pheasant/Hong Kong/FY155/01 (H5N1)) polysaccharase (PB1) gene, the part encoding sequence.

AF509174 influenza A virus (A/ Gallus Domesticus/Hong Kong/SF189/01 (H5N1)) polysaccharase (PB1) gene, the part encoding sequence.

AF509175 influenza A virus (A/ quail/Hong Kong/SF203/01 (H5N1)) polysaccharase (PB1) gene, the part encoding sequence.

AF509176 influenza A virus (A/ pigeon/Hong Kong/SF215/01 (H5N1)) polysaccharase (PB1) gene, the part encoding sequence.

AF509177 influenza A virus (A/ chicken/Hong Kong/SF219/01 (H5N1)) polysaccharase (PB1) gene, the part encoding sequence.

AF509178 influenza A virus (A/ chicken/Hong Kong/715.5/01 (H5N1)) polysaccharase (PB1) gene, the part encoding sequence.

AF509179 influenza A virus (A/ chicken/Hong Kong/751.1/01 (H5N1)) polysaccharase (PB1) gene, the part encoding sequence.

AF509180 influenza A virus (A/ chicken/Hong Kong/822.1/01 (H5N1)) polysaccharase (PB1) gene, the part encoding sequence.

AF509181 influenza A virus (A/ chicken/Hong Kong/829.2/01 (H5N1)) polysaccharase (PB1) gene, the part encoding sequence.

AF509182 influenza A virus (A/ chicken/Hong Kong/830.2/01 (H5N1)) polysaccharase (PB1) gene, the part encoding sequence.

AF509183 influenza A virus (A/ chicken/Hong Kong/858.3/01 (H5N1)) polysaccharase (PB1) gene, the part encoding sequence.

AF509184 influenza A virus (A/ chicken/Hong Kong/866.3/01 (H5N1)) polysaccharase (PB1) gene, the part encoding sequence.

AF509185 influenza A virus (A/ chicken/Hong Kong/867.1/01 (H5N1)) polysaccharase (PB1) gene, the part encoding sequence.

AF509186 influenza A virus (A/ chicken/Hong Kong/879.1/01 (H5N1)) polysaccharase (PB1) gene, the part encoding sequence.

AF509187 influenza A virus (A/ chicken/Hong Kong/873.3/01 (H5N1)) polysaccharase (PB1) gene, the part encoding sequence.

AF509188 influenza A virus (A/ chicken/Hong Kong/876.1/01 (H5N1)) polysaccharase (PB1) gene, the part encoding sequence.

AF509189 influenza A virus (A/ chicken/Hong Kong/891.1/01 (H5N1)) polysaccharase (PB1) gene, the part encoding sequence.

AF509190 influenza A virus (A/ chicken/Hong Kong/893.2/01 (H5N1)) polysaccharase (PB1) gene, the part encoding sequence.

AF509191 influenza A virus (A/ goose/Hong Kong/76.1/01 (H5N1)) polysaccharase (PB1) gene, the part encoding sequence.

AF509192 influenza A virus (A/ goose/Hong Kong/ww100/01 (H5N1)) polysaccharase (PB1) gene, the part encoding sequence.

AF509193 influenza A virus (A/ duck/Hong Kong/573.4/01 (H5N1)) polysaccharase (PB1) gene, the part encoding sequence.

AF509194 influenza A virus (A/ duck/Hong Kong/646.3/01 (H5N1)) polysaccharase (PB1) gene, the part encoding sequence.

AY035888 influenza A virus (A/ goose/Guangdong/3/97 (H5N1)) polysaccharase basic protein 1 (PB1) gene, encoding sequence completely.

AY059513 influenza A virus (A/ goose/Hong Kong/ww26/2000 (H5N1)) fragment 2 polysaccharases (PB1) gene, the part encoding sequence.

AY059514 influenza A virus (A/ goose/Hong Kong/ww28/2000 (H5N1)) fragment 2 polysaccharases (PB1) gene, the part encoding sequence.

AY059515 influenza A virus (A/ duck/Hong Kong/ww381/2000 (H5N1)) fragment 2 polysaccharases (PB1) gene, the part encoding sequence.

AY059516 influenza A virus (A/ duck/Hong Kong/ww461/2000 (H5N1)) fragment 2 polysaccharases (PB1) gene, the part encoding sequence.

AY059517 influenza A virus (A/ goose/Hong Kong/ww491/2000 (H5N1)) fragment 2 polysaccharases (PB1) gene, the part encoding sequence.

AY059518 influenza A virus (A/ duck/Hong Kong/2986.1/2000 (H5N1)) fragment 2 polysaccharases (PB1) gene, the part encoding sequence.

AY059519 influenza A virus (A/ goose/Hong Kong/3014.8/2000 (H5N1)) fragment 2 polysaccharases (PB1) gene, the part encoding sequence.

AY221575 influenza A virus (A/ chicken/Hong Kong/NT873.3/01-MB (H5N1)) polysaccharase basic protein 1 (PB1) gene, the part encoding sequence.

AY221576 influenza A virus (A/ chicken/Hong Kong/NT873.3/01 (H5N1)) polysaccharase basic protein 1 (PB1) gene, the part encoding sequence.

AY221577 influenza A virus (A/ chicken/Hong Kong/FY150/01-MB (H5N1)) polysaccharase basic protein 1 (PB1) gene, the part encoding sequence.

AY221578 influenza A virus (A/ chicken/Hong Kong/FY150/01 (H5N1)) polysaccharase basic protein 1 (PB1) gene, the part encoding sequence.

AY221579 influenza A virus (A/ pheasant/Hong Kong/FY155/01-MB (H5N1)) polysaccharase basic protein 1 (PB1) gene, the part encoding sequence.

AY221580 influenza A virus (A/ pheasant/Hong Kong/FY155/01 (H5N1)) polysaccharase basic protein 1 (PB1) gene, the part encoding sequence.

AY221581 influenza A virus (A/ chicken/Hong Kong/YU822.2/01-MB (H5N1)) polysaccharase basic protein 1 (PB1) gene, the part encoding sequence.

AY221582 influenza A virus (A/ chicken/Hong Kong/YU822.2/01 (H5N1)) polysaccharase basic protein 1 (PB1) gene, the part encoding sequence.

AY221583 influenza A virus (A/ chicken/Hong Kong/YU562/01 (H5N1)) polysaccharase basic protein 1 (PB1) gene, the part encoding sequence.

AY518366 influenza A virus (A/ duck/China/E319-2/03 (the H5N1)) PB1 of polysaccharase subunit (PB1) gene, encoding sequence completely.

AY576394 influenza A virus (A/Gs/ Hong Kong/739.2/02 (H5N1)) polysaccharase (PB1) gene, the part encoding sequence.

AY576395 influenza A virus (A/Eg/ Hong Kong/757.3/02 (H5N1)) polysaccharase (PB1) gene, the part encoding sequence.

AY576396 influenza A virus (A/G.H/ Hong Kong/793.1/02 (H5N1)) polysaccharase (PB1) gene, the part encoding sequence.

AY576397 influenza A virus (A/Dk/ Hong Kong/821/02 (H5N1)) polysaccharase (PB1) gene, encoding sequence completely.

AY576398 influenza A virus (A/Ck/ Hong Kong/31.4/02 (H5N1)) polysaccharase (PB1) gene, the part encoding sequence.

AY576399 influenza A virus (A/Ck/ Hong Kong/61.9/02 (H5N1)) polysaccharase (PB1) gene, the part encoding sequence.

AY576400 influenza A virus (A/Ck/ Hong Kong/YU777/02 (H5N1)) polysaccharase (PB1) gene, the part encoding sequence.

AY576401 influenza A virus (A/Ck/ Hong Kong/96.1/02 (H5N1)) polysaccharase (PB1) gene, the part encoding sequence.

AY576402 influenza A virus (A/Ck/ Hong Kong/409.1/02 (H5N1)) polysaccharase (PB1) gene, the part encoding sequence.

AY576403 influenza A virus (A/Ph/ Hong Kong/674.15/02 (H5N1)) polysaccharase (PB1) gene, the part encoding sequence.

AY585483 influenza A virus (A/ duck/Fujian/01/2002 (H5N1)) polysaccharase basic protein 1 (PB1) mRNA, encoding sequence completely.

AY585484 influenza A virus (A/ duck/Fujian/13/2002 (H5N1)) polysaccharase basic protein 1 (PB1) mRNA, encoding sequence completely.

AY585485 influenza A virus (A/ duck/Fujian/17/2001 (H5N1)) polysaccharase basic protein 1 (PB1) mRNA, encoding sequence completely.

AY585486 influenza A virus (A/ duck/Fujian/19/2000 (H5N1)) polysaccharase basic protein 1 (PB1) mRNA, encoding sequence completely.

AY585487 influenza A virus (A/ duck/Guangdong/01/2001 (H5N1)) polysaccharase basic protein 1 (PB1) mRNA, encoding sequence completely.

AY585488 influenza A virus (A/ duck/Guangdong/07/2000 (H5N1)) polysaccharase basic protein 1 (PB1) mRNA, encoding sequence completely.

AY585489 influenza A virus (A/ duck/Guangdong/12/2000 (H5N1)) polysaccharase basic protein 1 (PB1) mRNA, encoding sequence completely.

AY585490 influenza A virus (A/ duck/Guangdong/22/2002 (H5N1)) polysaccharase basic protein 1 (PB1) mRNA, encoding sequence completely.

AY585491 influenza A virus (A/ duck/Guangdong/40/2000 (H5N1)) polysaccharase basic protein 1 (PB1) mRNA, encoding sequence completely.

AY585492 influenza A virus (A/ duck/Guangxi/07/1999 (H5N1)) polysaccharase basic protein 1 (PB1) mRNA, encoding sequence completely.

AY585493 influenza A virus (A/ duck/Guangxi/22/2001 (H5N1)) polysaccharase basic protein 1 (PB1) mRNA, encoding sequence completely.

AY585494 influenza A virus (A/ duck/Guangxi/35/2001 (H5N1)) polysaccharase basic protein 1 (PB1) mRNA, encoding sequence completely.

AY585495 influenza A virus (A/ duck/Guangxi/50/2001 (H5N1)) polysaccharase basic protein 1 (PB1) mRNA, encoding sequence completely.

AY585496 influenza A virus (A/ duck/Guangxi/53/2002 (H5N1)) polysaccharase basic protein 1 (PB1) mRNA, encoding sequence completely.

AY585497 influenza A virus (A/ duck/Shanghai/08/2001 (H5N1)) polysaccharase basic protein 1 (PB1) mRNA, encoding sequence completely.

AY585498 influenza A virus (A/ duck/Shanghai/13/2001 (H5N1)) polysaccharase basic protein 1 (PB1) mRNA, encoding sequence completely.

AY585499 influenza A virus (A/ duck/Shanghai/35/2002 (H5N1)) polysaccharase basic protein 1 (PB1) mRNA, encoding sequence completely.

AY585500 influenza A virus (A/ duck/Shanghai/37/2002 (H5N1)) polysaccharase basic protein 1 (PB1) mRNA, encoding sequence completely.

AY585501 influenza A virus (A/ duck/Shanghai/38/2001 (H5N1)) polysaccharase basic protein 1 (PB1) mRNA, encoding sequence completely.

AY585502 influenza A virus (A/ duck/Zhejiang/11/2000 (H5N1)) polysaccharase basic protein 1 (PB1) mRNA, encoding sequence completely.

AY585503 influenza A virus (A/ duck/Zhejiang/52/2000 (H5N1)) polysaccharase basic protein 1 (PB1) mRNA, encoding sequence completely.

AY590582 influenza A virus (A/ chicken/Buddhist system/Thailand/CU-K2/2004 (H5N1)) polysaccharase basic protein 1 (PBP1) gene, encoding sequence completely.

AY609310 influenza A virus (A/ chicken/Guangdong/174/04 (H5N1)) fragment 2, sequence completely.

AY651651 influenza A virus (A/Ck/ Indonesia/BL/2003 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651652 influenza A virus (A/Dk/ Indonesia/MS/2004 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651653 influenza A virus (A/Ck/ Indonesia/PA/2003 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651654 influenza A virus (A/Ck/ Indonesia/4/2004 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651655 influenza A virus (A/Ck/ Indonesia/2A/2003 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651656 influenza A virus (A/Ck/ Indonesia/5/2004 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651657 influenza A virus (A/Ck/ Thailand/1/2004 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651658 influenza A virus (A/Ck/ Thailand/73/2004 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651659 influenza A virus (A/Ck/ Thailand/9.1/2004 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651660 influenza A virus (A/Qa/ Thailand/57/2004 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651661 influenza A virus (A/bird/ Thailand/3.1/2004 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651662 influenza A virus (A/Dk/ Thailand/71.1/2004 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651663 influenza A virus (A/Gs/ Thailand/79/2004 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651668 influenza A virus (A/Ck/ Vietnam/33/2004 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651669 influenza A virus (A/Ck/ Vietnam/35/2004 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651670 influenza A virus (A/Ck/ Vietnam/36/2004 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651671 influenza A virus (A/Ck/ Vietnam/37/2004 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651672 influenza A virus (A/Ck/ Vietnam/38/2004 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651673 influenza A virus (A/Ck/ Vietnam/39/2004 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651674 influenza A virus (A/Ck/ Vietnam/C57/2004 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651675 influenza A virus (A/Dk/ Vietnam/11/2004 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651676 influenza A virus (A/Gf/ Hong Kong/38/2002 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651677 influenza A virus (A/Ck/ Hong Kong/31.2/2002 (H5N1)) polysaccharase alkalescence subunit 1 gene, the part encoding sequence.

AY651678 influenza A virus (A/Ck/ Hong Kong/37.4/2002 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651679 influenza A virus (A/SCk/ Hong Kong/YU100/2002 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651680 influenza A virus (A/Ck/ Hong Kong/YU22/2002 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651681 influenza A virus (A/Ck/ Hong Kong/3176.3/2002 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651682 influenza A virus (A/Ck/ Hong Kong/3169.1/2002 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651683 influenza A virus (A/Ck/ Hong Kong/FY157/2003 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651684 influenza A virus (A/Ck/ Hong Kong/YU324/2003 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651685 influenza A virus (A/Ck/ Hong Kong/2133.1/2003 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651686 influenza A virus (A/Ck/ Hong Kong/NT93/2003 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651687 influenza A virus (A/Ck/ Hong Kong/SSP141/2003 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651688 influenza A virus (A/Ck/ Hong Kong/WF157/2003 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651689 influenza A virus (the red mouth of A/ gull/Hong Kong/12.1/2003 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651690 influenza A virus (A/ rock dove/Hong Kong/862.7/2002 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651691 influenza A virus (A/ heron/Hong Kong/861.1/2002 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651692 influenza A virus (A/ sets sparrow/Hong Kong/864/2002 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651693 influenza A virus (A/ teal/China/2978.1/2002 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651694 influenza A virus (A/ falcon/Hong Kong/D0028/2004 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651695 influenza A virus (A/Dk/HN/5806/2003 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651696 influenza A virus (A/Dk/ST/4003/2003 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651697 influenza A virus (A/Ck/ST/4231/2003 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651698 influenza A virus (A/Dk/YN/6255/2003 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651699 influenza A virus (A/Dk/YN/6445/2003 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651700 influenza A virus (A/Ph/ST/44/2004 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651701 influenza A virus (A/Dk/HN/303/2004 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651702 influenza A virus (A/Dk/HN/101/2004 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651703 influenza A virus (A/Ck/YN/374/2004 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY651704 influenza A virus (A/Ck/YN/115/2004 (H5N1)) polysaccharase alkalescence subunit 1 (PB1) gene, the part encoding sequence.

AY653199 influenza A virus (A/ chicken/Jilin/9/2004 (H5N1)) fragment 2, sequence completely.

AY676025 influenza A virus strain (A/ duck/Hong Kong/821/02 (H5N1)) polysaccharase basic protein 1 (PB1) gene, encoding sequence completely.

AY676026 influenza A virus strain (A/ egression/Hong Kong/757.2/03 (H5N1)) polysaccharase basic protein 1 (PB1) gene, encoding sequence completely.

AY676027 influenza A virus strain (A/ chicken/Korea S/ES/03 (H5N1)) polysaccharase basic protein 1 (PB1) gene, encoding sequence completely.

AY676028 influenza A virus strain (A/ duck/Korea S/ESD1/03 (H5N1)) polysaccharase basic protein 1 (PB1) gene, coding row completely.

AY684704 influenza A virus (A/ chicken/Hubei/327/2004 (H5N1)) polysaccharase basic protein 1 (PB1) gene, encoding sequence completely.

AY737287 influenza A virus (A/ chicken/Guangdong/191/04 (H5N1)) fragment 2, sequence completely.

AY737294 influenza A virus (A/ chicken/Guangdong/178/04 (H5N1)) fragment 2, sequence completely.

AY737302 influenza A virus (A/ duck/Guangdong/173/04 (H5N1)) fragment 2, sequence completely.

AY770083 influenza A virus (A/ chicken/Hubei/489/2004 (H5N1)) non-functional polysaccharase basic protein 1 (PB1) gene, sequence completely.

AY770994 influenza A virus (A/ chicken/big subtlety/Thailand/CU-23/04 (H5N1)) polysaccharase basic protein 1 gene, the part encoding sequence.

AY818130 influenza A virus (A/ chicken/Vietnam/C58/04 (H5N1)) polymerase protein PB1 gene, encoding sequence completely.

AY818131 influenza A virus (A/ quail/Vietnam/36/04 (H5N1)) polymerase protein PB1 gene, encoding sequence completely.

AY856862 influenza A virus (A/ duck/Shandong/093/2004 (H5N1)) fragment 2, sequence completely.

AB188822 influenza A virus (A/ chicken/capital of a country/3/2004 (H5N1)) polysaccharase basic protein 1 (PB1) gene, encoding sequence completely.

AB189051 influenza A virus (A/ crow/capital of a country/53/2004 (H5N1)) polysaccharase basic protein 1 (PB1) gene, encoding sequence completely.

AB189060 influenza A virus (A/ crow/Osaka/102/2004 (H5N1)) polysaccharase basic protein 1 (PB1) PB1 gene, encoding sequence completely.

AF046085 influenza A virus (A/ chicken/Hong Kong/220/97 (H5N1)) polysaccharase basic protein 1 (PB1) gene, encoding sequence completely.

AF098590 influenza A virus (A/ chicken/Hong Kong/258/97 (H5N1)) PB1 albumen (PB1) gene, the part encoding sequence.

AF098591 influenza A virus (A/ chicken/Hong Kong/y388/97 (H5N1)) PB1 albumen (PB1) gene, the part encoding sequence.

AF098592 influenza A virus (A/ chicken/Hong Kong/728/97 (H5N1)) PBI albumen (PB1) gene, the part encoding sequence.

AF098593 influenza A virus (A/ chicken/Hong Kong/786/97 (H5N1)) PB1 albumen (PB1) gene, the part encoding sequence.

AF098594 influenza A virus (A/ chicken/Hong Kong/915/97 (H5N1)) PB1 albumen (PB1) gene, the part encoding sequence.

AF098595 influenza A virus (A/ duck/Hong Kong/p46/97 (H5N1)) PB1 albumen (PB1) gene, the part encoding sequence.

AF098596 influenza A virus (A/ duck/Hong Kong/y283/97 (H5N1)) PB1 albumen (PB1) gene, the part encoding sequence.

AF098598 influenza A virus (A/ goose/Hong Kong/w355/97 (H5N1)) PB1 albumen (PB1) gene, the part encoding sequence.

AF144301 influenza A virus (A/ goose/Guangdong/1/96 (H5N1)) polysaccharase (PB1) gene, encoding sequence completely.

AF216716 influenza A virus (A/ environment/Hong Kong/437-4/99 (H5N1)) polysaccharase basic protein 1 gene, encoding sequence completely.

AF216724 influenza A virus (A/ environment/Hong Kong/437-6/99 (H5N1)) polysaccharase basic protein 1 gene, encoding sequence completely.

AF216732 influenza A virus (A/ environment/Hong Kong/437-8/99 (H5N1)) polysaccharase basic protein 1 gene, encoding sequence completely.

AF216740 influenza A virus (A/ environment/Hong Kong/437-10/99 (H5N1)) polysaccharase basic protein 1 gene, encoding sequence completely.

AY303663 influenza A virus (A/ chicken/Chile/176822/02 (H7N3)) polysaccharase basic protein 1 gene, encoding sequence completely.

AY303664 influenza A virus (A/ chicken/Chile/4957/02 (H7N3)) polysaccharase basic protein 1 gene, the part encoding sequence.

AY586435 influenza A virus (A/ turkey/Italy/214845/02 (H7N3)) PB1 gene, the part encoding sequence.

AY586436 influenza A virus (A/ turkey/Italy/220158/2002 (H7N3)) PB1 gene, the part encoding sequence.

AY586437 influenza A virus (A/ wild duck/Italy/33/01 (H7N3)) PB1 gene, the part encoding sequence.

AY586438 influenza A virus (A/ wild duck/Italy/43/01 (H7N3)) PBI gene, the part encoding sequence.

AY616765 influenza A virus (A/ chicken/British Columbia/04 (H7N3)) PB1 polysaccharase subunit (PB1) gene, encoding sequence completely.

AY646084 influenza A virus (A/ chicken/British Columbia/GSC_ people _ B/04 (H7N3)) polysaccharase basic protein 1 (PB1) gene, encoding sequence completely.

AY648293 influenza A virus (A/GSC_ chicken _ B/ British Columbia/04 (H7N3)) PB1 polysaccharase subunit (PB1) gene, encoding sequence completely.

AY653039 influenza A virus (A/GSC_ chicken/British Columbia/04 (H7N3)) PB1 polysaccharase subunit (PB1) gene, encoding sequence completely.

AJ620348 influenza A virus (A/ chicken/Germany/R28/03 (H7N7)) RNA polymerase PB1 gene, geneome RNA.

AY340080 influenza A virus (A/ Holland/124/03 (H7N7)) polysaccharase (PB1) gene, the part encoding sequence.

AY340081 influenza A virus (A/ Holland/126/03 (H7N7)) polysaccharase (PB1) gene, the part encoding sequence.

AY340082 influenza A virus (A/ Holland/127/03 (H7N7)) polysaccharase (PB1) gene, the part encoding sequence.

AY340083 influenza A virus (A/ Holland/219/03 (H7N7)) polysaccharase (PB1) gene, encoding sequence completely.

AY340084 influenza A virus (A/ Holland/033/03 (H7N7)) polysaccharase (PB1) gene, encoding sequence completely.

AY340085 influenza A virus (A/ chicken/Holland/1/03 (H7N7)) polysaccharase (PB1) gene, encoding sequence completely.

AB049155 influenza A virus (A/ parakeet/Chiba/1/97 (H9N2)) PB1 gene for polysaccharase basic protein 1, encoding sequence completely.

AB049156 influenza A virus (A/ parakeet/Narita Airport/92A/98 (H9N2)) polysaccharase basic protein 1 (PB1) gene, encoding sequence completely.

AF508618 influenza A virus (A/ ostrich/South Africa/9508103/95 (H9N2)) fragment 2 polysaccharase PB1 (PB1) genes, the part encoding sequence.

AF508619 influenza A virus (A/ chicken/Pakistan/4/99 (H9N2)) fragment 2 polysaccharase PB1 (PB1) genes, the part encoding sequence.

AF508620 influenza A virus (A/ chicken/Pakistan/5/99 (H9N2)) fragment 2 polysaccharase PB1 (PB1) genes, the part encoding sequence.

AF508621 influenza A virus (A/ chicken/Germany/R45/98 (H9N2)) fragment 2 polysaccharase PB1 (PB1) genes, the part encoding sequence.

AF508622 influenza A virus (A/ duck/Germany/113/95 (H9N2)) fragment 2 polysaccharase PB1 (PB1) genes, the part encoding sequence.

AF508623 influenza A virus (A/ chicken/Iran/11T/99 (H9N2)) fragment 2 polysaccharase PB1 (PB1) genes, the part encoding sequence.

AF508624 influenza A virus (A/ chicken/Saudi Arabia/532/99 (H9N2)) fragment 2 polysaccharase PB1 (PB1) genes, the part encoding sequence.

AF508625 influenza A virus (A/ pheasant/Ireland/PV18/97 (H9N2)) fragment 2 polysaccharase PBI (PB1) genes, the part encoding sequence.

AF508626 influenza A virus (A/ chicken/Korea S/99029/99 (H9N2)) fragment 2 polysaccharase PB1 (PB1) genes, the part encoding sequence.

AF508627 influenza A virus (A/ chicken/Beijing/8/98 (H9N2)) fragment 2 polysaccharase PB1 (PB1) gene, encoding sequences completely.

AF508628 influenza A virus (A/ chicken/Guangdong/10/00 (H9N2)) fragment 2 polysaccharase PB1 (PB1) genes, the part encoding sequence.

AF508629 influenza A virus (A/ chicken/Guangdong/11/97 (H9N2)) fragment 2 polysaccharase PB1 (PB1) gene, encoding sequences completely.

AF508630 influenza A virus (A/ chicken/Hebei/4/98 (H9N2)) fragment 2 polysaccharase PB1 (PB1) gene, encoding sequences completely.

AF508631 influenza A virus (A/ chicken/Heilungkiang/10/97 (H9N2)) fragment 2 polysaccharase PB1 (PB1) genes, the part encoding sequence.

AF508632 influenza A virus (A/ chicken/Henan/62/00 (H9N2)) fragment 2 polysaccharase PB1 (PB1) gene, encoding sequences completely.

AF508633 influenza A virus (A/ chicken/Ningxia/5/99 (H9N2)) fragment 2 polysaccharase PB1 (PB1) gene, encoding sequences completely.

AF508634 influenza A virus (A/ chicken/Sichuan/5/97 (H9N2)) fragment 2 polysaccharase PB1 (PB1) genes, the part encoding sequence.

AF508635 influenza A virus (A/ chicken/Shandong/6/96 (H9N2)) fragment 2PB1 (PB1) gene, the part encoding sequence.

AF508636 influenza A virus (A/ chicken/Shijiazhuang/2/99 (H9N2)) fragment 2 polysaccharase PB1 (PB1) gene, encoding sequences completely.

AF508637 influenza A virus (A/ chicken/Shenzhen/9/97 (H9N2)) fragment 2 polysaccharase PB1 (PB1) gene, encoding sequences completely.

AF508638 influenza A virus (A/ duck/Nanjing/1/97 (H9N2)) fragment 2 polysaccharase PB1 (PB1) gene, encoding sequences completely.

AF508639 influenza A virus (A/ quail/Shanghai/8/96 (H9N2)) fragment 2 polysaccharase PB1 (PB1) gene, encoding sequences completely.

AF523427 influenza A virus (A/ duck/Shantou/830/00 (H9N2)) polysaccharase (PB1) gene, the part encoding sequence.

AF523428 influenza A virus (A/ duck/Shantou/2102/00 (H9N2)) polysaccharase (PB1) gene, the part encoding sequence.

AF523429 influenza A virus (A/ duck/Shantou/1043/00 (H9N2)) polysaccharase (PB1) gene, the part encoding sequence.

AF523430 influenza A virus (A/ duck/Shantou/2134/00 (H9N2)) polysaccharase (PB1) gene, the part encoding sequence.

AF523431 influenza A virus (A/ wild duck/Shantou/4808/01 (H9N2)) polysaccharase (PB1) gene, the part encoding sequence.

AF523432 influenza A virus (A/ duck/Shantou/2144/00 (H9N2)) polysaccharase (PB1) gene, the part encoding sequence.

AF523433 influenza A virus (A/ duck/Shantou/2143/00 (H9N2)) polysaccharase (PB1) gene, the part encoding sequence.

AF523434 influenza A virus (A/ duck/Shantou/1796/00 (H9N2)) polysaccharase (PB1) gene, the part encoding sequence.

AF523435 influenza A virus (A/ duck/Shantou/2088/01 (H9N2)) polysaccharase (PB1) gene, the part encoding sequence.

AF523436 influenza A virus (A/ duck/Shantou/1881/00 (H9N2)) polysaccharase (PB1) gene, the part encoding sequence.

AF523437 influenza A virus (A/ duck/Hong Kong/366/78 (H9N2)) polysaccharase (PB1) gene, the part encoding sequence.

AF523438 influenza A virus (A/ duck/Hong Kong/552/79 (H9N2)) polysaccharase (PB1) gene, the part encoding sequence.

AF523439 influenza A virus (A/ duck/Hong Kong/86/76 (H9N2)) polysaccharase (PB1) gene, the part encoding sequence.

AF523440 influenza A virus (A/ duck/Hong Kong/289/78 (H9N2)) polysaccharase (PB1) gene, the part encoding sequence.

AF523441 influenza A virus (A/ duck/Hong Kong/610/79 (H9N2)) polysaccharase (PB1) gene, the part encoding sequence.

AF523442 influenza A virus (A/ duck/Shantou/1605/01 (H9N2)) polysaccharase (PB1) gene, the part encoding sequence.

AF523443 influenza A virus (A/ duck/Shantou/1042/00 (H9N2)) polysaccharase (PB1) gene, the part encoding sequence.

AF536659 influenza A virus (A/ chicken/Beijing/1/95 (H9N2)) PB1 gene, the part encoding sequence.

AF536660 influenza A virus (A/ chicken/Beijing/2/97 (H9N2)) PB1 gene, the part encoding sequence.

AF536661 influenza A virus (A/ chicken/Beijing/3/99 (H9N2)) PB1 gene, the part encoding sequence.

AF536662 influenza A virus (A/ chicken/Guangdong/97 (H9N2)) PB1 gene, the part encoding sequence.

AF536663 influenza A virus (A/ chicken/Hebei/1/96 (H9N2)) PB1 gene, the part encoding sequence.

AF536664 influenza A virus (A/ chicken/Hebei/2/98 (H9N2)) PB1 gene, the part encoding sequence.

AF536665 influenza A virus (A/ chicken/Hebei/3/98 (H9N2)) PBI gene, the part encoding sequence.

AF536666 influenza A virus (A/ chicken/Henan/98 (H9N2)) PB1 gene, the part encoding sequence.

AF536667 influenza A virus (A/ chicken/Liaoning/99 (H9N2)) PBI gene, the part encoding sequence.

AF536668 influenza A virus (A/ chicken/Shandong/98 (H9N2)) PB1 gene, the part encoding sequence.

AJ291396 influenza A virus (A/ chicken/Pakistan/2/99 (H9N2)) polysaccharase PB1 (PB1) gene, geneome RNA.

AJ427862 influenza A virus (A/ quail/Hong Kong/FY298/00 (H9N2)) part PB1 polymerase protein pb1 gene, geneome RNA

AY180840 influenza A virus strain A/ pigeon/Nanchang/7-058/2000 (H9N2) the polysaccharase PB1 of subunit (PB1) gene, the part encoding sequence.

AY180843 influenza A virus strain A/ quail/Nanchang/2-0460/2000 (H9N2) the polysaccharase PB1 of subunit (PB1) gene, the part encoding sequence.

AY180844 influenza A virus strain A/ pigeon/Nanchang/2-0461/2000 (H9N2) the polysaccharase PB1 of subunit (PB1) gene, the part encoding sequence.

AY180851 influenza A virus strain A/ pigeon/Nanchang/11-145/2000 (H9N2) the polysaccharase PB1 of subunit (PB1) gene, the part encoding sequence.

AY180852 influenza A virus strain A/ duck/Nanchang/11-197/2000 (H9N2) the polysaccharase PB1 of subunit (PB1) gene, the part encoding sequence.

AY180854 influenza A virus strain A/ duck/Nanchang/11-290/2000 (H9N2) the polysaccharase PB1 of subunit (PB1) gene, the part encoding sequence.

AY180856 influenza A virus strain A/ duck/Nanchang/1-0070/2000 (H9N2) the polysaccharase PB1 of subunit (PB1) gene, the part encoding sequence.

AY180866 influenza A virus strain A/ duck/Nanchang/7-092/2000 (H9N2) the polysaccharase PB1 of subunit (PB1) gene, the part encoding sequence.

AY180867 influenza A virus strain A/ chicken/Nanchang/1-0016/2000 (H9N2) the polysaccharase PB1 of subunit (PB1) gene, the part encoding sequence.

AY180873 influenza A virus strain A/ chicken/Nanchang/4-010/2000 (H9N2) the polysaccharase PB1 of subunit (PB1) gene, the part encoding sequence.

AY180874 influenza A virus strain A/ chicken/Nanchang/4-301/2001 (H9N2) the polysaccharase PB1 of subunit (PB1) gene, the part encoding sequence.

AY180875 influenza A virus strain A/ chicken/Nanchang/4-361/2001 (H9N2) the polysaccharase PB1 of subunit (PB1) gene, the part encoding sequence.

AY180892 influenza A virus strain A/ quail/Nanchang/4-040/2000 (H9N2) the polysaccharase PB1 of subunit (PB1) gene, the part encoding sequence.

AY180897 influenza A virus strain A/Wild duck/Nanchang/2-0480/2000 (H9N2) the polysaccharase PB1 of subunit (PB1) gene, the part encoding sequence.

AY180900 influenza A virus strain A/ duck/Nanchang/10-389/2000 (H9N2) the polysaccharase PB1 of subunit (PB1) gene, the part encoding sequence.

AY180901 influenza A virus strain A/ duck/Nanchang/11-392/2000 (H9N2) the polysaccharase PB1 of subunit (PB1) gene, the part encoding sequence.

AY253751 influenza A virus (A/ chicken/Shanghai/F/98 (H9N2)) polysaccharase basic protein 1 (PB1) gene, encoding sequence completely.

AY307947 influenza A virus (A/ chicken/Beijing/1/00 (H9N2)) polysaccharase subunit (PB1) gene, the part encoding sequence.

AY307948 influenza A virus (A/ chicken/Hebei/1/01 (H9N2)) polysaccharase subunit (PB1) gene, the part encoding sequence.

The P1 of RNA polymerase subunit (PB1) gene that AY633170 influenza A virus (A/ wild duck/alberta is economized/17/91 (H9N2)) RNA instructs, the part encoding sequence.

The P1 of RNA polymerase subunit (PB1) gene that AY633282 influenza A virus (A/ wild duck/alberta is economized/321/88 (H9N2)) RNA instructs, the part encoding sequence.

The P1 of RNA polymerase subunit (PB1) gene that AY633298 influenza A virus (A/ wild duck/alberta is economized/11/91 (H9N2)) RNA instructs, the part encoding sequence.

AY664774 influenza A virus (A/ chicken/Hong Kong/CSW153/03 (H9N2)) polysaccharase basic protein 1 (PB1) gene, the part encoding sequence.

AY664775 influenza A virus (A/ chicken/Hong Kong/AP45/03 (H9N2)) polysaccharase basic protein 1 (PB1) gene, the part encoding sequence.

AY664776 influenza A virus (A/ chicken/Hong Kong/BD90/03 (H9N2)) polysaccharase basic protein 1 (PB1) gene, the part encoding sequence.

AY664777 influenza A virus (A/ chicken/Hong Kong/CSW291/03 (H9N2)) polysaccharase basic protein 1 (PB1) gene, the part encoding sequence.

AY664778 influenza A virus (A/ chicken/Hong Kong/CSW304/03 (H9N2)) polysaccharase basic protein 1 (PB1) gene, the part encoding sequence.

AY664779 influenza A virus (A/ chicken/Hong Kong/FY23/03 (H9N2)) polysaccharase basic protein 1 (PB1) gene, the part encoding sequence.

AY664780 influenza A virus (A/ vulture galeeny/Hong Kong/NT101/03 (H9N2)) polysaccharase basic protein 1 (PB1) gene, the part encoding sequence.

AY664781 influenza A virus (A/ chicken/Hong Kong/NT142/03 (H9N2)) polysaccharase basic protein 1 (PB1) gene, the part encoding sequence.

AY664782 influenza A virus (A/ chicken/Hong Kong/SF1/03 (H9N2)) polysaccharase basic protein 1 (PB1) gene, the part encoding sequence.

AY664783 influenza A virus (A/ chicken/Hong Kong/SSP101/03 (H9N2)) polysaccharase basic protein 1 (PB1) gene, the part encoding sequence.

AY664784 influenza A virus (A/ chicken/Hong Kong/TP38/03 (H9N2)) polysaccharase basic protein 1 (PB1) gene, the part encoding sequence.

AY664785 influenza A virus (A/ chicken/Hong Kong/WF126/03 (H9N2)) polysaccharase basic protein 1 (PB1) gene, the part encoding sequence.

AY664786 influenza A virus (A/ pigeon/Hong Kong/WF53/03 (H9N2)) polysaccharase basic protein 1 (PB1) gene, the part encoding sequence.

AY664787 influenza A virus (A/ pheasant/Hong Kong/WF54/03 (H9N2)) polysaccharase basic protein 1 (PB1) gene, the part encoding sequence.

AY664788 influenza A virus (A/ vulture galeeny/Hong Kong/NT184/03 (H9N2)) polysaccharase basic protein 1 (PB1) gene, the part encoding sequence.

AY664789 influenza A virus (A/ chicken/Hong Kong/WF120/03 (H9N2)) polysaccharase basic protein 1 (PB1) gene, the part encoding sequence.

AY664790 influenza A virus (A/ chicken/Hong Kong/NT366/03 (H9N2)) polysaccharase basic protein 1 (PB1) gene, the part encoding sequence.

AY664791 influenza A virus (A/ chicken/Hong Kong/YU427/03 (H9N2)) polysaccharase basic protein 1 (PB1) gene, the part encoding sequence.

AY800239 influenza A virus (A/ chicken/Korea S/S1/2003 (H9N2)) polysaccharase basic protein 1 (PB1) gene, the part encoding sequence.

AY862694 influenza A virus (A/ bamboo silk chicken (silky chicken)/Korea S/S3/03 (H9N2)) PB1 (PB1) gene, the part encoding sequence.

AY862695 influenza A virus (A/ chicken/Korea S/S4/03 (H9N2)) PB1 (PB1) gene, the part encoding sequence.

AY862696 influenza A virus (A/ chicken/Korea S/S5/03 (H9N2)) PB1 (PB1) gene, the part encoding sequence.

AY862697 influenza A virus (A/ chicken/Korea S/S12/03 (H9N2)) PB1 (PB1) gene, the part encoding sequence.

AY862698 influenza A virus (A/ duck/Korea S/S13/03 (H9N2)) PB1 (PB1) gene, the part encoding sequence.

AY862699 influenza A virus (A/ dove/Korea S/S14/03 (H9N2)) PB1 (PB1) gene, the part encoding sequence.

AY862700 influenza A virus (A/ chicken/Korea S/S15/03 (H9N2)) PB1 (PB1) gene, the part encoding sequence.

AY862701 influenza A virus (A/ chicken/Korea S/S16/03 (H9N2)) PBI (PB1) gene, the part encoding sequence.

AY862702 influenza A virus (A/ chicken/Korea S/S18/03 (H9N2)) PB1 (PB1) gene, the part encoding sequence.

AF156416 influenza A virus (A/ chicken/Hong Kong/G9/97 (H9N2)) fragment 2 PB1 polysaccharase subunit (PB1) genes, the part encoding sequence.

AF156417 influenza A virus (A/ chicken/Hong Kong/G23/99 (H9N2)) fragment 2 PB1 polysaccharase subunit (PB1) genes, the part encoding sequence.

AF156418 influenza A virus (A/ pigeon/Hong Kong/Y233/97 (H9N2)) fragment 2 PB1 polysaccharase subunit (PB1) genes, the part encoding sequence.

AF156419 influenza A virus (A/ duck/Hong Kong/Y280/97 (H9N2)) fragment 2 PB1 polysaccharase subunit (PB1) genes, the part encoding sequence.

AF156420 influenza A virus (A/ duck/Hong Kong/Y439/97 (H9N2)) fragment 2 PB1 polysaccharase subunit (PB1) genes, the part encoding sequence.

AF156421 influenza A virus (A/ quail/Hong Kong/G1/97 (H9N2)) fragment 2 PB1 polysaccharase subunit (PB1) genes, the part encoding sequence.

AF156422 influenza A virus (A/ chicken/Hong Kong/739/94 (H9N2)) fragment 2 PB1 polysaccharase subunit (PB1) genes, the part encoding sequence.

AF156423 influenza A virus (A/ chicken/Beijing/1/94 (H9N2)) fragment 2 PB1 polysaccharase subunit (PB1) genes, the part encoding sequence.

AF156424 influenza A virus (A/ quail/Hong Kong/AF157/92 (H9N2)) fragment 2 PB1 polysaccharase subunit (PB1) genes, the part encoding sequence.

AF156425 influenza A virus (A/ chicken/Korea S/38349-p96323/96 (H9N2)) fragment 2 PB1 polysaccharase subunit (PB1) genes, the part encoding sequence.

AF156426 influenza A virus (A/ chicken/Korea S/25232-96006/96 (H9N2)) fragment 2 PB1 polysaccharase subunit (PB1) genes, the part encoding sequence.

AF156427 influenza A virus (A/ bank bird/Delaware/9/96 (H9N2)) fragment 2 PB1 polysaccharase subunit (PB1) genes, the part encoding sequence.

AF156428 influenza A virus (A/ quail/Arkansas State/29209-1/93 (H9N2)) fragment 2 PB1 polysaccharase subunit (PB1) genes, the part encoding sequence.

AF156429 influenza A virus (A/ turkey/California/189/66 (H9N2)) fragment 2 PB1 polysaccharase subunit (PB1) genes, the part encoding sequence.

AF222632 influenza A virus (A/ quail/Hong Kong/A17/99 (H9N2)) fragment 2 polysaccharase 1 (PB1) genes, the part encoding sequence.

AF222633 influenza A virus (A/ pigeon/Hong Kong/FY6/99 (H9N2)) fragment 2 polysaccharase 1 (PB1) genes, the part encoding sequence.

AF222634 influenza A virus (A/ chicken/Hong Kong/NT16/99 (H9N2)) fragment 2 polysaccharase 1 (PB1) genes, the part encoding sequence.

AF222635 influenza A virus (A/ quail/Hong Kong/SSP10/99 (H9N2)) fragment 2 polysaccharase 1 (PB1) genes, the part encoding sequence.

AF222636 influenza A virus (A/ pheasant/Hong Kong/SSP11/99 (H9N2)) fragment 2 polysaccharase 1 (PB1) genes, the part encoding sequence.

AF222637 influenza A virus (A/ chicken/Hong Kong/FY20/99 (H9N2)) fragment 2 polysaccharase 1 (PB1) genes, the part encoding sequence.

AF222638 influenza A virus (A/ chicken/Hong Kong/KC12/99 (H9N2)) fragment 2 polysaccharase 1 (PB1) genes, the part encoding sequence.

AF222639 influenza A virus (A/ quail/Hong Kong/NT2899 (H9N2)) fragment 2 polysaccharase 1 (PB1) genes, the part encoding sequence.

AF222640 influenza A virus (A/ chicken/Hong Kong/SF2/99 (H9N2)) fragment 2 polysaccharase 1 (PB1) genes, the part encoding sequence.

AF222641 influenza A virus (A/Silky chicken/Hong Kong/SF44/99 (H9N2)) fragment 2 polysaccharase 1 (PB1) genes, the part encoding sequence.

Used sequence during influenza A polysaccharase basic protein 2 (PB2) is analyzed

Gi|49356919|AY633219| influenza A virus (A/ wild duck/alberta is economized/211/98 (H1N1)) polysaccharase basic protein 2 (PB2) gene, the part encoding sequence.

Gi|27466107|AY180748| influenza A virus strain A/ quail/Nanchang/12-340/2000 (H1N1) the polysaccharase PB2 of subunit (PB2) gene, the part encoding sequence.

Gi|45272173|AY422042| influenza A virus (A/ duck/Hokkaido/95/01 (H2N2)) polysaccharase subunit (PB2) gene, the part encoding sequence.

Gi|18091825|AF213910| influenza A virus (A/ chicken/Italy/5945/95 (H3N2)) fragment 1PB2 polymerase protein gene, the part encoding sequence.

Gi|27466133|AY180761| influenza A virus strain A/ chicken/Nanchang/3-120/2001 (H3N2) the polysaccharase PB2 of subunit (PB2) gene, the part encoding sequence.

Gi|56160002|AY779267| influenza A virus (the A/ turkey/state North Carolina state/12344/03 (H3N2)) polysaccharase basic protein 2 (PB2) gene, the part encoding sequence.

Gi|56160004|AY779268| influenza A virus (A/ turkey/Minnesota State/764-2/03 (H3N2)) polysaccharase basic protein 2 (PB2) gene, the part encoding sequence.

Gi|58429704|AY862719| influenza A virus (A/ chicken/Korea S/S6/03 (H3N2)) PB2 (PB2) gene, the part encoding sequence.

Gi|58429706|AY862720| influenza A virus (A/ duck/Korea S/S7/03 (H3N2)) PB2 (PB2) gene, the part encoding sequence.

Gi|58429708|AY862721| influenza A virus (A/ duck/Korea S/S8/03 (H3N2)) PB2 (PB2) gene, the part encoding sequence.

Gi|58429710|AY862722| influenza A virus (A/ duck/Korea S/S9/03 (H3N2)) PB2 (PB2) gene, the part encoding sequence.

Gi|58429712|AY862723| influenza A virus (A/ duck/Korea S/S10/03 (H3N2)) PB2 (PB2) gene, the part encoding sequence.

Gi|58429714|AY862724| influenza A virus (A/ dove/Korea S/S11/03 (H3N2)) PB2 (PB2) gene, the part encoding sequence.

Gi|5805276|AF144300| influenza A virus (A/ goose/Guangdong/1/96 (H5N1)) polysaccharase (PB2) gene, encoding sequence completely.

Gi|3335416|AF046086| influenza A virus (A/ chicken/Hong Kong/220/97 (H5N1)) polysaccharase basic protein 2 (PB2) gene, the part encoding sequence.

Gi|6048841|AF098577| influenza A virus (A/ chicken/Hong Kong/258/97 (H5N1)) PB2 albumen (PB2) gene, the part encoding sequence.

Gi|6048843|AF098578| influenza A virus (A/ chicken/Hong Kong/y388/97 (H5N1)) PB2protein (PB2) gene, the part encoding sequence.

Gi|6048845|AF098579| influenza A virus (A/ chicken/Hong Kong/728/97 (H5N1)) PB2 albumen (PB2) gene, the part encoding sequence.

Gi|6048847|AF098580| influenza A virus (A/ chicken/Hong Kong/786/97 (H5N1)) PB2 albumen (PB2) gene, the part encoding sequence.

Gi|6048849|AF098581| influenza A virus (A/ chicken/Hong Kong/915/97 (H5N1)) PB2 albumen (PB2) gene, the part encoding sequence.

Gi|6048851|AF098582| influenza A virus (A/ duck/Hong Kong/p46/97 (H5N1)) PB2 albumen (PB2) gene, the part encoding sequence.

Gi|6048853|AF098583| influenza A virus (A/ duck/Hong Kong/y283/97 (H5N1)) PB2 albumen (PB2) gene, the part encoding sequence.

Gi|6048855|AF098584| influenza A virus (A/ goose/Hong Kong/w355/97 (H5N1)) PB2 albumen (PB2) gene, the part encoding sequence.

Gi|14860983|AY038798| influenza A virus (A/ goose/Guangdong/3/1997 (H5N1)) PB2 albumen (PB2) gene, encoding sequence completely.

Gi|47156244|AY585513| influenza A virus (A/ duck/Guangxi/07/1999 (H5N1)) polysaccharase basic protein 2 (PB2) mRNA, encoding sequence completely.

Gi|9863884|AF216717| influenza A virus (A/ environment/Hong Kong/437-4/99 (H5N1)) polysaccharase basic protein 2 genes, the part encoding sequence.

Gi|9863903|AF216725| influenza A virus (A/ environment/Hong Kong/437-6/99 (H5N1)) polysaccharase basic protein 2 genes, the part encoding sequence.

Gi|9863921|AF216733| influenza A virus (A/ environment/Hong Kong/437-8/99 (H5N1)) polysaccharase basic protein 2 genes, the part encoding sequence.

Gi|9863939|AF216741| influenza A virus (A/ environment/Hong Kong/437-10/99 (H5N1)) polysaccharase basic protein 2 genes, the part encoding sequence.

Gi|47156264|AY585523| influenza A virus (A/ duck/Zhejiang/11/2000 (H5N1)) polysaccharase basic protein 2 (PB2) mRNA, encoding sequence completely.

Gi|47156266|AY585524| influenza A virus (A/ duck/Zhejiang/52/2000 (H5N1)) polysaccharase basic protein 2 (PB2) mRNA, encoding sequence completely.

Gi|47156232|AY585507| influenza A virus (A/ duck/Fujian/19/2000 (H5N1)) polysaccharase basic protein 2 (PB2) mRNA, encoding sequence completely.

Gi|47156236|AY585509| influenza A virus (A/ duck/Guangdong/07/2000 (H5N1)) polysaccharase basic protein 2 (PB2) mRNA, encoding sequence completely.

Gi|47156238|AY585510| influenza A virus (A/ duck/Guangdong/12/2000 (H5N1)) polysaccharase basic protein 2 (PB2) mRNA, encoding sequence completely.

Gi|47156242|AY585512| influenza A virus (A/ duck/Guangdong/40/2000 (H5N1)) polysaccharase basic protein 2 (PB2) mRNA, encoding sequence completely.

Gi|19697849|AY059520| influenza A virus (A/ goose/Hong Kong/ww26/2000 (H5N1)) fragment 1 polysaccharase (PB2) gene, the part encoding sequence.

Gi|19697851|AY059521| influenza A virus (A/ goose/Hong Kong/ww28/2000 (H5N1)) fragment 1 polysaccharase (PB2) gene, the part encoding sequence.

Gi|19697853|AY059522| influenza A virus (A/ duck/Hong Kong/ww381/2000 (H5N1)) fragment 1 polysaccharase (PB2) gene, the part encoding sequence.

Gi|19697855|AY059523| influenza A virus (A/ duck/Hong Kong/ww461/2000 (H5N1)) fragment 1 polysaccharase (PB2) gene, the part encoding sequence.

Gi|19697857|AY059524| influenza A virus (A/ goose/Hong Kong/ww491/2000 (H5N1)) fragment 1 polysaccharase (PB2) gene, the part encoding sequence.

Gi|19697859|AY059525| influenza A virus (A/ duck/Hong Kong/2986.1/2000 (H5N1)) fragment 1 polysaccharase (PB2) gene, the part encoding sequence.

Gi|19697867|AY059529| influenza A virus (A/ goose/Hong Kong/3014.8/2000 (H5N1)) fragment 1 polysaccharase (PB2) gene, the part encoding sequence.

Gi|18092181|AF398425| influenza A virus (A/ goose/Hong Kong/385.3/2000 (H5N1)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|18092183|AF398426| influenza A virus (A/ goose/Hong Kong/385.5/2000 (H5N1)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|21359665|AF468840| influenza A virus (A/ duck/Anyang/AVL-1/2001 (H5N1)) polysaccharase basic protein 2 (PB2) gene, the part encoding sequence.

Gi|28849606|AF509143| influenza A virus (A/ chicken/Hong Kong/FY77/01 (H5N1)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|28849608|AF509144| influenza A virus (A/ chicken/Hong Kong/YU562/01 (H5N1)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|28849610|AF509145| influenza A virus (A/ chicken/Hong Kong/YU563/01 (H5N1)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|28849612|AF509146| influenza A virus (A/ chicken/Hong Kong/FY150/01 (H5N1)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|28849614|AF509147| influenza A virus (A/ pheasant/Hong Kong/FY155/01 (H5N1)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|28849616|AF509148| influenza A virus (A/ bamboo silk chicken/Hong Kong/SF189/01 (H5N1)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|28849618|AF509149| influenza A virus (A/ quail/Hong Kong/SF203/01 (H5N1)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|28849620|AF509150| influenza A virus (A/ pigeon/Hong Kong/SF215/01 (H5N1)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|28849622|AF509151| influenza A virus (A/ chicken/Hong Kong/SF219/01 (H5N1)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|28849624|AF509152| influenza A virus (A/ chicken/Hong Kong/715.5/01 (H5N1)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|28849626|AF509153| influenza A virus (A/ chicken/Hong Kong/751.1/01 (H5N1)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|28849628|AF509154| influenza A virus (A/ chicken/Hong Kong/822.1/01 (H5N1)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|28849630|AF509155| influenza A virus (A/ chicken/Hong Kong/829.2/01 (H5N1)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|28849632|AF509156| influenza A virus (A/ chicken/Hong Kong/830.2/01 (H5N1)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|28849634|AF509157| influenza A virus (A/ chicken/Hong Kong/858.3/01 (H5N1)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|28849636|AF509158| influenza A virus (A/ chicken/Hong Kong/866.3/01 (H5N1)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|28849638|AF509159| influenza A virus (A/ chicken/Hong Kong/867.1/01 (H5N1)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|28849640|AF509160| influenza A virus (A/ chicken/Hong Kong/879.1/01 (H5N1)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|28849642|AF509161| influenza A virus (A/ chicken/Hong Kong/873.3/01 (H5N1)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|28849644|AF509162| influenza A virus (A/ chicken/Hong Kong/876.1/01 (H5N1)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|28849646|AF509163| influenza A virus (A/ chicken/Hong Kong/891.1/01 (H5N1)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|28849648|AF509164| influenza A virus (A/ chicken/Hong Kong/893.2/01 (H5N1)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|28849650|AF509165| influenza A virus (A/ goose/Hong Kong/76.1/01 (H5N1)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|28849652|AF509166| influenza A virus (A/ goose/Hong Kong/ww100/01 (H5N1)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|28849654|AF509167| influenza A virus (A/ duck/Hong Kong/573.4/01 (H5N1)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|28849656|AF509168| influenza A virus (A/ duck/Hong Kong/646.3/01 (H5N1)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|28823262|AY221584| influenza A virus (A/ chicken/Hong Kong/NT873.3/01-MB (H5N1)) polysaccharase basic protein 2 (PB2) gene, the part encoding sequence.

Gi|28823443|AY221585| influenza A virus (A/ chicken/Hong Kong/NT873.3/01 (H5N1)) polysaccharase basic protein 2 (PB2) gene, the part encoding sequence.

Gi|28823612|AY221586| influenza A virus (A/ chicken/Hong Kong/FY150/01-MB (H5N1)) polysaccharase basic protein 2 (PB2) gene, the part encoding sequence.

Gi|28823783|AY221587| influenza A virus (A/ chicken/Hong Kong/FY150/01 (H5N1)) polysaccharase basic protein 2 (PB2) gene, the part encoding sequence.

Gi|28823961|AY221588| influenza A virus (A/ pheasant/Hong Kong/FY155/01-MB (H5N1)) polysaccharase basic protein 2 (PB2) gene, the part encoding sequence.

Gi|28824143|AY221589| influenza A virus (A/ pheasant/Hong Kong/FY155/01 (H5N1)) polysaccharase basic protein 2 (PB2) gene, the part encoding sequence.

Gi|28824334|AY221590| influenza A virus (A/ chicken/Hong Kong/YU822.2/01-MB (H5N1)) polysaccharase basic protein 2 (PB2) gene, the part encoding sequence.

Gi|28824502|AY221591| influenza A virus (A/ chicken/Hong Kong/YU822.2/01 (H5N1)) polysaccharase basic protein 2 (PB2) gene, the part encoding sequence.

Gi|28824684|AY221592| influenza A virus (A/ chicken/Hong Kong/YU562/01 (H5N1)) polysaccharase basic protein 2 (PB2) gene, the part encoding sequence.

Gi|47156230|AY585506| influenza A virus (A/ duck/Fujian/17/2001 (H5N1)) polysaccharase basic protein 2 (PB2) mRNA, encoding sequence completely.

Gi|47156234|AY585508| influenza A virus (A/ duck/Guangdong/01/2001 (H5N1)) polysaccharase basic protein 2 (PB2) mRNA, encoding sequence completely.

Gi|47156246|AY585514| influenza A virus (A/ duck/Guangxi/22/2001 (H5N1)) polysaccharase basic protein 2 (PB2) mRNA, encoding sequence completely.

Gi|47156248|AY585515| influenza A virus (A/ duck/Guangxi/35/2001 (H5N1)) polysaccharase basic protein 2 (PB2) mRNA, encoding sequence completely.

Gi|47156250|AY585516| influenza A virus (A/ duck/Guangxi/50/2001 (H5N1)) polysaccharase basic protein 2 (PB2) mRNA, encoding sequence completely.

Gi|47156254|AY585518| influenza A virus (A/ duck/Shanghai/08/2001 (H5N1)) polysaccharase basic protein 2 (PB2) mRNA, encoding sequence completely.

Gi|47156256|AY585519| influenza A virus (A/ duck/Shanghai/13/2001 (H5N1)) polysaccharase basic protein 2 (PB2) mRNA, encoding sequence completely.

Gi|47156262|AY585522| influenza A virus (A/ duck/Shanghai/38/2001 (H5N1)) polysaccharase basic protein 2 (PB2) mRNA, encoding sequence completely.

Gi|47156258|AY585520| influenza A virus (A/ duck/Shanghai/35/2002 (H5N1)) polysaccharase basic protein 2 (PB2) mRNA, encoding sequence completely.

Gi|47156260|AY585521| influenza A virus (A/ duck/Shanghai/37/2002 (H5N1)) polysaccharase basic protein 2 (PB2) mRNA, encoding sequence completely.

Gi|47156252|AY585517| influenza A virus (A/ duck/Guangxi/53/2002 (H5N1)) polysaccharase basic protein 2 (PB2) mRNA, encoding sequence completely.

Gi|47156240|AY585511| influenza A virus (A/ duck/Guangdong/22/2002 (H5N1)) polysaccharase basic protein 2 (PB2) mRNA, encoding sequence completely.

Gi|47834791|AY576382| influenza A virus (A/Gs/ Hong Kong/739.2/02 (H5N1)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|47834793|AY576383| influenza A virus (A/Eg/ Hong Kong/757.3/02 (H5N1)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|47834795|AY576384| influenza A virus (A/G.H/ Hong Kong/793.1/02 (H5N1)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|47834797|AY576385| influenza A virus (A/Dk/ Hong Kong/821/02 (H5N1)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|47834799|AY576386| influenza A virus (A/Ck/ Hong Kong/31.4/02 (H5N1)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|47834801|AY576387| influenza A virus (A/Ck/ Hong Kong/61.9/02 (H5N1)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|47834803|AY576388| influenza A virus (A/Ck/ Hong Kong/YU777/02 (H5N1)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|47834805|AY576389| influenza A virus (A/Ck/ Hong Kong/96.1/02 (H5N1)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|47834807|AY576390| influenza A virus (A/Ck/ Hong Kong/409.1/02 (H5N1)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|47834809|AY576391| influenza A virus (A/Ph/ Hong Kong/sv674.15/02 (H5N1)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|47156226|AY585504| influenza A virus (A/ duck/Fujian/01/2002 (H5N1)) polysaccharase basic protein 2 (PB2) mRNA, encoding sequence completely.

Gi|47156228|AY585505| influenza A virus (A/ duck/Fujian/13/2002 (H5N1)) polysaccharase basic protein 2 (PB2) mRNA, encoding sequence completely.

Gi|50296597|AY651744| influenza A virus (A/ heron/Hong Kong/861.1/2002 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, the part encoding sequence.

Gi|50296599|AY651745| influenza A virus (A/ rock dove/Hong Kong/862.7/2002 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, the part encoding sequence.

Gi|50296601|AY651746| influenza A virus (A/ sets sparrow/Hong Kong/864/2002 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, the part encoding sequence.

Gi|50296603|AY651747| influenza A virus (A/ teal/China/2978.1/2002 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, the part encoding sequence.

Gi|56548879|AY676021| influenza A virus (A/ duck/Hong Kong/821/02 (H5N1)) polysaccharase basic protein 2 (PB2) gene, encoding sequence completely.

Gi|50296569|AY651730| influenza A virus (A/Gf/ Hong Kong/38/2002 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, the part encoding sequence.

Gi|50296571|AY651731| influenza A virus (A/Ck/ Hong Kong/31.2/2002 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, the part encoding sequence.

Gi|50296573|AY651732| influenza A virus (A/Ck/ Hong Kong/37.4/2002 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, the part encoding sequence.

Gi|50296575|AY651733| influenza A virus (A/SCk/ Hong Kong/YU100/2002 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, the part encoding sequence.

Gi|50296577|AY651734| influenza A virus (A/Ck/ Hong Kong/YU22/2002 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, the part encoding sequence.

Gi|50296579|AY651735| influenza A virus (A/Ck/ Hong Kong/3176.3/2002 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, the part encoding sequence.

Gi|50296581|AY651736| influenza A virus (A/Ck/ Hong Kong/3169.1/2002 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, the part encoding sequence.

Gi|50296583|AY651737| influenza A virus (A/Ck/ Hong Kong/FY157/2003 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, the part encoding sequence.

Gi|50296585|AY651738| influenza A virus (A/Ck/ Hong Kong/YU324/2003 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, the part encoding sequence.

Gi|50296587|AY651739| influenza A virus (A/Ck/ Hong Kong/2133.1/2003 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, the part encoding sequence.

Gi|50296589|AY651740| influenza A virus (A/Ck/ Hong Kong/NT93/2003 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, the part encoding sequence.

Gi|50296591|AY651741| influenza A virus (A/Ck/ Hong Kong/SSP141/2003 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, the part encoding sequence.

Gi|50296593|AY651742| influenza A virus (A/Ck/ Hong Kong/WF157/2003 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, the part encoding sequence.

Gi|50296595|AY651743| influenza A virus (the red mouth of A/ gull/Hong Kong/12.1/2003 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, the part encoding sequence.

Gi|50296607|AY651749| influenza A virus (A/Dk/HN/5806/2003 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, the part encoding sequence.

Gi|50296609|AY651750| influenza A virus (A/Dk/ST/4003/2003 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, the part encoding sequence.

Gi|50296611|AY651751| influenza A virus (A/Ck/ST/4231/2003 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, the part encoding sequence.

Gi|50296613|AY651752| influenza A virus (A/Dk/YN/6255/2003 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, the part encoding sequence.

Gi|50296615|AY651753| influenza A virus (A/Dk/YN/6445/2003 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, the part encoding sequence.

Gi|50296529|AY651710| influenza A virus (A/Ck/ Indonesia/2A/2003 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, the part encoding sequence.

Gi|56548881|AY676022| influenza A virus (A/ egression/Hong Kong/757.2/03 (H5N1)) polysaccharase basic protein 2 (PB2) gene, encoding sequence completely.

Gi|56548883|AY676023| influenza A virus (A/ chicken/Korea S/ES/03 (H5N1)) polysaccharase basic protein 2 (PB2) gene, encoding sequence completely.

Gi|56548885|AY676024| influenza A virus (A/ duck/Korea S/ESD1/03 (H5N1)) polysaccharase basic protein 2 (PB2) gene, encoding sequence completely.

Gi|50296519|AY651705| influenza A virus (A/Ck/ Indonesia/PA/2003 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, encoding sequence completely.

Gi|50296523|AY651707| influenza A virus (A/Ck/ Indonesia/BL/2003 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, encoding sequence completely.

Gi|41207501|AY518367| influenza A virus (A/ duck/China/E319-2/03 (the H5N1)) PB2 of polysaccharase subunit (PB2) gene, encoding sequence completely.

Gi|45359369|AY550147| influenza A virus (A/ chicken/Buddhist system/Thailand/CU-K2/04 (H5N1)) polysaccharase basic protein 2 (PB2) gene, the part encoding sequence.

Gi|50296525|AY651708| influenza A virus (A/Ck/ Indonesia/5/2004 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, the part encoding sequence.

Gi|50296527|AY651709| influenza A virus (A/Ck/ Indonesia/4/2004 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, the part encoding sequence.

Gi|50296521|AY651706| influenza A virus (A/Dk/ Indonesia/MS/2004 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, encoding sequence completely.

Gi|51094103|AY590581| influenza A virus (A/ chicken/Buddhist system/Thailand/CU-K2/2004 (H5N1)) polysaccharase basic protein 2 (PBP2) gene, the part encoding sequence.

Gi|47716766|AY609309| influenza A virus (A/ chicken/Guangdong/174/04 (H5N1)) fragment 1, sequence completely.

Gi|58531082|AB166859| influenza A virus (A/ chicken/mountain pass/7/2004 (H5N1)) PB2 gene for polysaccharase basic protein 2, encoding sequence completely.

The PB2 gene of gi|58531114|AB188813| influenza A virus (A/ chicken/big/8/2004 (H5N1) that divide) polysaccharase basic protein 2, encoding sequence completely.

Gi|50956621|AY684703| influenza A virus (A/ chicken/Hubei/327/2004 (H5N1)) polysaccharase basic protein 2 (PB2) gene, encoding sequence completely.

Gi|57915957|AY737286| influenza A virus (A/ chicken/Guangdong/191/04 (H5N1)) fragment 1, sequence completely.

Gi|57916006|AY737293| influenza A virus (A/ chicken/Guangdong/178/04 (H5N1)) fragment 1, sequence completely.

Gi|57916060|AY737301| influenza A virus (A/ duck/Guangdong/173/04 (H5N1)) fragment 1, sequence completely.

Gi|55233237|AY770084| influenza A virus (A/ chicken/Hubei/489/2004 (H5N1)) polysaccharase basic protein 2 (PB2) gene, encoding sequence completely.

Gi|54873461|AY770993| influenza A virus (A/ chicken/big subtlety (Ayutthaya)/Thailand/CU-23/04 (H5N1)) polysaccharase basic protein 2 genes, the part encoding sequence.

Gi|58618421|AY818127| influenza A virus (A/ chicken/Vietnam/C58/04 (H5N1)) polymerase protein PB2 gene, encoding sequence completely.

Gi|58618423|AY818128| influenza A virus (A/ quail/Vietnam/36/04 (H5N1)) polymerase protein PB2 gene, encoding sequence completely.

Gi|58374183|AY856861| influenza A virus (A/ duck/Shandong/093/2004 (H5N1)) fragment 1, sequence completely.

The PB2 gene of gi|58531132|AB188821| influenza A virus (A/ chicken/capital of a country/3/2004 (H5N1)) polysaccharase basic protein 2, encoding sequence completely.

The PB2 gene of gi|58531150|AB189050| influenza A virus (A/ crow/capital of a country/53/2004 (H5N1)) polysaccharase basic protein 2, encoding sequence completely.

The PB2 gene of gi|58531168|AB189058| influenza A virus (A/ crow/Osaka/102/2004 (H5N1)) polysaccharase basic protein 2, encoding sequence completely.

Gi|50296605|AY651748| influenza A virus (A/ falcon/Hong Kong/D0028/2004 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, encoding sequence completely.

Gi|50296531|AY651711| influenza A virus (A/Ck/ Thailand/1/2004 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, the part encoding sequence.

Gi|50296533|AY651712| influenza A virus (A/Ck/ Thailand/73/2004 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, the part encoding sequence.

Gi|50296535|AY651713| influenza A virus (A/Ck/ Thailand/9.1/2004 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, encoding sequence completely.

Gi|50296537|AY651714| influenza A virus (A/Qa/ Thailand/57/2004 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, encoding sequence completely.

Gi|50296539|AY651715| influenza A virus (A/bird/ Thailand/3.1/2004 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, encoding sequence completely.

Gi|50296541|AY651716| influenza A virus (A/Dk/ Thailand/71.1/2004 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, encoding sequence completely.

Gi|50296543|AY651717| influenza A virus (A/Gs/ Thailand/79/2004 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, the part encoding sequence.

Gi|50296553|AY651722| influenza A virus (A/Ck/ Vietnam/33/2004 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, encoding sequence completely.

Gi|50296555|AY651723| influenza A virus (A/Ck/ Vietnam/35/2004 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, encoding sequence completely.

Gi|50296557|AY651724| influenza A virus (A/Ck/ Vietnam/36/2004 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, encoding sequence completely.

Gi|50296559|AY651725| influenza A virus (A/Ck/ Vietnam/37/2004 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, encoding sequence completely.

Gi|50296561|AY651726| influenza A virus (A/Ck/ Vietnam/38/2004 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, encoding sequence completely.

Gi|50296563|AY651727| influenza A virus (A/Ck/ Vietnam/39/2004 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, encoding sequence completely.

Gi|50296565|AY651728| influenza A virus (A/Ck/ Vietnam/C57/2004 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, encoding sequence completely.

Gi|50296567|AY651729| influenza A virus (A/Dk/ Vietnam/11/2004 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, encoding sequence completely.

Gi|50296617|AY651754| influenza A virus (A/Ck/YN/374/2004 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, the part encoding sequence.

Gi|50296619|AY651755| influenza A virus (A/Ck/YN/115/2004 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, the part encoding sequence.

Gi|50296621|AY651756| influenza A virus (A/Ph/ST/44/2004 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, the part encoding sequence.

Gi|50296623|AY651757| influenza A virus (A/Dk/HN/303/2004 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, the part encoding sequence.

Gi|50296625|AY651758| influenza A virus (A/Dk/HN/101/2004 (H5N1)) polysaccharase alkalescence subunit 2 (PB2) gene, the part encoding sequence.

Gi|50365712|AY653193| influenza A virus (A/ chicken/Jilin/9/2004 (H5N1)) fragment 1, sequence completely.

Gi|47680940|AY586445| influenza A virus (A/ wild duck/Italy/43/01 (H7N3)) PB2 gene, the part encoding sequence.

Gi|47680930|AY586440| influenza A virus (A/ wild duck/Italy/33/01 (H7N3)) PB2 gene, the part encoding sequence.

Gi|47680932|AY586441| influenza A virus (A/ turkey/Italy/220158/2002 (H7N3)) PB2 gene, the part encoding sequence.

The PB2 gene of gi|45124743|AJ627485| influenza A virus (A/ turkey/Italy/214845/2002 (H7N3)) RNA polymerase, geneome RNA.

The PB2 gene of gi|45124767|AJ627496| influenza A virus (A/ turkey/Italy/220158/2002 (H7N3)) RNA polymerase, geneome RNA.

Gi|34597782|AY303665| influenza A virus (A/ chicken/Chile/176822/02 (H7N3)) polysaccharase basic protein 2 genes, the part encoding sequence.

Gi|34597784|AY303666| influenza A virus (A/ chicken/Chile/4957/02 (H7N3)) polysaccharase basic protein 2 genes, the part encoding sequence.

Gi|47680928|AY586439| influenza A virus (A/ turkey/Italy/214845/02 (H7N3)) PB2 gene, the part encoding sequence.

Gi|47834374|AY616766| influenza A virus (A/ chicken/British Columbia/04 (H7N3)) PB2 polysaccharase subunit (PB2) gene, encoding sequence completely.

Gi|50542651|AY646085| influenza A virus (A/ chicken/British Columbia/GSC_ people _ B/04 (H7N3)) polysaccharase basic protein 2 (PB2) gene, encoding sequence completely.

Gi|50083053|AY648294| influenza A virus (A/GSC_ chicken _ B/ British Columbia/04 (H7N3)) PB2 polysaccharase subunit (PB2) gene, encoding sequence completely.

Gi|50059194|AY650276| influenza A virus (A/GSC_ chicken/British Columbia/04 (H7N3)) PB2 polysaccharase subunit (PB2) gene, encoding sequence completely.

Gi|60700|X58691| influenza A virus (the m cap binding protein of the A/FPV/ nymph of the dragonfly/27 (H7N7) PB2 gene, geneome RNA

Gi|325001|M38291| influenza A virus/FPV/Weybridge polysaccharase basic protein 2 (PB2) (fragment 3) gene, encoding sequence completely.

Gi|9988661|AF268120| influenza A virus (the red abdomen dunlin of A/ (RedKnot)/Delaware/259/94 (H7N7)) polymerase protein PB2 gene, the part encoding sequence.

The PB2 gene of gi|40732893|AJ620347| influenza A virus ((A/ chicken/Germany/R28/03 (H7N7)) A/ chicken/Germany/R28/03 (H7N7)) RNA polymerase, geneome RNA.

Gi|37813157|AY342410| influenza A virus (A/ Holland/124/03 (H7N7)) polymerase protein 2 genes, the part encoding sequence.

Gi|37813159|AY342411| influenza A virus (A/ Holland/126/03 (H7N7)) polymerase protein 2 genes, the part encoding sequence.

Gi|37813161|AY342412| influenza A virus (A/ Holland/127/03 (H7N7)) polymerase protein 2 genes, the part encoding sequence.

Gi|37813163|AY342413| influenza A virus (A/ Holland/219/03 (H7N7)) polymerase protein 2 genes, the part encoding sequence.

Gi|37813165|AY342414| influenza A virus (A/ chicken/Holland/1/03 (H7N7)) polymerase protein 2 genes, the part encoding sequence.

Gi|5732354|AF156443| influenza A virus (A/ turkey/California/189/66 (H9N2)) fragment 1PB2 polysaccharase subunit (PB2) gene, the part encoding sequence.

Gi|31339587|AF523469| influenza A virus (A/ duck/Hong Kong/86/76 (H9N2)) polymerase protein (PB2) gene, the part encoding sequence.

Gi|31339583|AF523467| influenza A virus (A/ duck/Hong Kong/366/78 (H9N2)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|31339605|AF523478| influenza A virus (A/ duck/Hong Kong/289/78 (H9N2)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|31339585|AF523468| influenza A virus (A/ duck/Hong Kong/552/79 (H9N2)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|31339589|AF523470| influenza A virus (A/ duck/Hong Kong/610/79 (H9N2)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|49356935|AY633283| influenza A virus (A/ wild duck/alberta is economized/321/88 (H9N2)) polysaccharase basic protein 2 (PB2) gene, the part encoding sequence.

Gi|49356939|AY633299| influenza A virus (A/ wild duck/alberta is economized/11/91 (H9N2)) polysaccharase basic protein 2 (PB2) gene, the part encoding sequence.

Gi|49356907|AY633171| influenza A virus (A/ wild duck/alberta is economized/17/91 (H9N2)) polysaccharase basic protein 2 (PB2) gene, the part encoding sequence.

Gi|5732342|AF156437| influenza A virus (A/ quail/Hong Kong/AF157/92 (H9N2)) fragment 1 PB2 polysaccharase subunit (PB2) gene, the part encoding sequence.

Gi|5732352|AF156442| influenza A virus (A/ quail/Arkansas State/29209-1/93 (H9N2)) fragment 1 PB2 polysaccharase subunit (PB2) gene, the part encoding sequence.

Gi|5732340|AF156436| influenza A virus (A/ chicken/Hong Kong/739/94 (H9N2)) fragment 1 PB2 polysaccharase subunit (PB2) gene, the part encoding sequence.

Gi|5732344|AF156438| influenza A virus (A/ chicken/Beijing/1/94 (H9N2)) fragment 1 PB2 polysaccharase subunit (PB2) gene, the part encoding sequence.

Gi|22759060|AF536679| influenza A virus (A/ chicken/Beijing/1/95 (H9N2)) PB2 gene, the part encoding sequence.

Gi|33318110|AF508640| influenza A virus (A/ ostrich/South Africa/9508103/95 (H9N2)) fragment 1 polysaccharase PB2 (PB2) gene, encoding sequence completely.

Gi|33318118|AF508644| influenza A virus (A/ duck/Germany/113/95 (H9N2)) fragment 1 polysaccharase PB2 (PB2) gene, the part encoding sequence.

Gi|33318144|AF508657| influenza A virus (A/ chicken/Shandong/6/96 (H9N2)) fragment 1 polysaccharase PB2 (PB2) gene, the part encoding sequence.

Gi|33318152|AF508661| influenza A virus (A/ quail/Shanghai/8/96 (H9N2)) fragment 1 polysaccharase PB2 (PB2) gene, the part encoding sequence.

Gi|5732346|AF156439| influenza A virus (A/ chicken/Korea S/38349-p96323/96 (H9N2)) fragment 1 PB2 polysaccharase subunit (PB2) gene, the part encoding sequence.

Gi|5732348|AF156440| influenza A virus (A/ chicken/Korea S/25232-96006/96 (H9N2)) fragment 1 PB2 polysaccharase subunit (PB2) gene, the part encoding sequence.

Gi|5732350|AF156441| influenza A virus (A/ bank bird/Delaware/9/96 (H9N2)) fragment 1 PB2 polysaccharase subunit (PB2) gene, the part encoding sequence.

Gi|22759068|AF536683| influenza A virus (A/ chicken/Hebei/1/96 (H9N2)) PB2 gene, the part encoding sequence.

Gi|5732328|AF156430| influenza A virus (A/ chicken/Hong Kong/G9/97 (H9N2)) fragment 1 PB2 polysaccharase subunit (PB2) gene, encoding sequence completely.

Gi|5732330|AF156431| influenza A virus (A/ chicken/Hong Kong/G23/97 (H9N2)) fragment 1 PB2 polysaccharase subunit (PB2) gene, the part encoding sequence.

Gi|5732332|AF156432| influenza A virus (A/ pigeon/Hong Kong/Y233/97 (H9N2)) fragment 1 PB2 polysaccharase subunit (PB2) gene, the part encoding sequence.

Gi|5732334|AF156433| influenza A virus (A/ duck/Hong Kong/Y280/97 (H9N2)) fragment 1 PB2 polysaccharase subunit (PB2) gene, the part encoding sequence.

Gi|5732336|AF156434| influenza A virus (A/ duck/Hong Kong/Y439/97 (H9N2)) fragment 1 PB2 polysaccharase subunit (PB2) gene, the part encoding sequence.

Gi|5732338|AF156435| influenza A virus (A/ quail/Hong Kong/G1/97 (H9N2)) fragment 1 PB2 polysaccharase subunit (PB2) gene, the part encoding sequence.

Gi|33318148|AF508659| influenza A virus (A/ chicken/Shenzhen/9/97 (H9N2)) fragment 1 polysaccharase PB2 (PB2) gene, the part encoding sequence.

Gi|33318150|AF508660| influenza A virus (A/ duck/Nanjing/1/97 (H9N2)) fragment 1 polysaccharase PB2 (PB2) gene, the part encoding sequence.

Gi|33318124|AF508647| influenza A virus (A/ pheasant/Ireland/PV18/97 (H9N2)) fragment 1 polysaccharase PB2 (PB2) gene, the part encoding sequence.

The PB2 gene of gi|13383266|AB049153| influenza A virus (A/ parakeet/Chiba/1/97 (H9N2)) polysaccharase basic protein 2, encoding sequence completely.

Gi|33318132|AF508651| influenza A virus (A/ chicken/Guangdong/11/97 (H9N2)) fragment 1 polysaccharase PB2 (PB2) gene, the part encoding sequence.

Gi|33318136|AF508653| influenza A virus (A/ chicken/Heilungkiang/10/97 (H9N2)) fragment 1 polysaccharase PB2 (PB2) gene, the part encoding sequence.

Gi|22759062|AF536680| influenza A virus (A/ chicken/Beijing/2/97 (H9N2)) PB2 gene, the part encoding sequence.

Gi|33318142|AF508656| influenza A virus (A/ chicken/Sichuan/5/97 (H9N2)) fragment 1 polysaccharase PB2 (PB2) gene, the part encoding sequence.

Gi|22759066|AF536682| influenza A virus (A/ chicken/Guangdong/97 (H9N2)) PB2 gene, the part encoding sequence.

Gi|22759078|AF536688| influenza A virus (A/ chicken/Shandong/98 (H9N2)) PB2 gene, the part encoding sequence.

Gi|33318116|AF508643| influenza A virus (A/ chicken/Germany/R45/98 (H9N2)) fragment 1 polysaccharase PB2 (PB2) gene, the part encoding sequence.

Gi|33318134|AF508652| influenza A virus (A/ chicken/Hebei/4/98 (H9N2)) fragment 1 polysaccharase PB2 (PB2) gene, the part encoding sequence.

Gi|33318128|AF508649| influenza A virus (A/ chicken/Beijing/8/98 (H9N2)) fragment 1 polysaccharase PB2 (PB2) gene, encoding sequence completely.

The PB2 gene of gi|13383268|AB049154| influenza A virus (A/ parakeet/Narita Airport/92A/98 (H9N2)) polysaccharase basic protein 2, encoding sequence completely.

Gi|22759070|AF536684| influenza A virus (A/ chicken/Hebei/2/98 (H9N2)) PB2 gene, the part encoding sequence.

Gi|22759072|AF536685| influenza A virus (A/ chicken/Hebei/3/98 (H9N2)) PB2 gene, the part encoding sequence.

Gi|22759074|AF536686| influenza A virus (A/ chicken/Henan/98 (H9N2)) PB2 gene, the part encoding sequence.

Gi|30025722|AY253750| influenza A virus (A/ chicken/Shanghai/F/98 (H9N2)) RNA polymerase (PB2) gene, encoding sequence completely.

Gi|12060631|AF222622| influenza A virus (A/ quail/Hong Kong/A17/99 (H9N2)) fragment 1 polysaccharase 2 (PB2) gene, the part encoding sequence.

Gi|12060633|AF222623| influenza A virus (A/ pigeon/Hong Kong/FY6/99 (H9N2)) fragment 1 polysaccharase 2 (PB2) gene, the part encoding sequence.

Gi|12060635|AF222624| influenza A virus (A/ chicken/Hong Kong/NT16/99 (H9N2)) fragment 1 polysaccharase 2 (PB2) gene, the part encoding sequence.

Gi|12060637|AF222625| influenza A virus (A/ quail/Hong Kong/SSP10/99 (H9N2)) fragment 1 polysaccharase 2 (PB2) gene, the part encoding sequence.

Gi|12060639|AF222626| influenza A virus (A/ pheasant/Hong Kong/SSP11/99 (H9N2)) fragment 1 polysaccharase 2 (PB2) gene, the part encoding sequence.

Gi|12060641|AF222627| influenza A virus (A/ chicken/Hong Kong/FY20/99 (H9N2)) fragment 1 polysaccharase 2 (PB2) gene, the part encoding sequence.

Gi|12060643|AF222628| influenza A virus (A/ chicken/Hong Kong/KC12/99 (H9N2)) fragment 1 polysaccharase 2 (PB2) gene, the part encoding sequence.

Gi|12060645|AF222629| influenza A virus (A/ quail/Hong Kong/NT28/99 (H9N2)) fragment 1 polysaccharase 2 (PB2) gene, the part encoding sequence.

Gi|12060647|AF222630| influenza A virus (A/ chicken/Hong Kong/SF2/99 (H9N2)) fragment 1 polysaccharase 2 (PB2) gene, the part encoding sequence.

Gi|12060649|AF222631| influenza A virus (A/Silky chicken/Hong Kong/SF44/99 (H9N2)) fragment 1 polysaccharase 2 (PB2) gene, the part encoding sequence.

Gi|22759076|AF536687| influenza A virus (A/ chicken/Liaoning/99 (H9N2)) PB2 gene, the part encoding sequence.

Gi|33318112|AF508641| influenza A virus (A/ chicken/Pakistan/4/99 (H9N2)) fragment 1 polysaccharase PB2 (PB2) gene, encoding sequence completely.

Gi|33318114|AF508642| influenza A virus (A/ chicken/Pakistan/5/99 (H9N2)) fragment 1 polysaccharase PB2 (PB2) gene, encoding sequence completely.

Gi|33318126|AF508648| influenza A virus (A/ chicken/Korea S/99029/99 (H9N2)) fragment 1 polysaccharase PB2 (PB2) gene, the part encoding sequence.

Gi|33318120|AF508645| influenza A virus (A/ chicken/Iran/11T/99 (H9N2)) fragment 1 polysaccharase PB2 (PB2) gene, encoding sequence completely.

Gi|33318122|AF508646| influenza A virus (A/ chicken/Saudi Arabia/532/99 (H9N2)) fragment 1 polysaccharase PB2 (PB2) gene, the part encoding sequence.

Gi|33318140|AF508655| influenza A virus (A/ chicken/Ningxia/5/99 (H9N2)) fragment 1 polysaccharase PB2 (PB2) gene, the part encoding sequence.

Gi|33318146|AF508658| influenza A virus (A/ chicken/Shijiazhuang/2/99 (H9N2)) fragment 1 polysaccharase PB2 (PB2) gene, the part encoding sequence.

Gi|12038893|AJ291395| influenza A virus (A/ chicken/Pakistan/2/99 (H9N2)) polysaccharase PB2 gene, geneome RNA.

Gi|22759064|AF536681| influenza A virus (A/ chicken/Beijing/3/99 (H9N2)) PB2 gene, the part encoding sequence.

Gi|31339607|AF523479| influenza A virus (A/ duck/Shantou/1881/00 (H9N2)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|31339601|AF523476| influenza A virus (A/ duck/Shantou/830/00 (H9N2)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|31339603|AF523477| influenza A virus (A/ duck/Shantou/1796/00 (H9N2)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|18496117|AJ427861| influenza A virus (A/ quail/Hong Kong/FY298/00 (H9N2)) PB2 polysaccharase part pb2 gene, geneome RNA

Gi|27466041|AY180715| influenza A virus strain A/ wild duck/Nanchang/2-0480/2000 (H9N2) the polysaccharase PB2 of subunit (PB2) gene, the part encoding sequence.

Gi|27466043|AY180716| influenza A virus strain A/ pigeon/Nanchang/2-0461/2000 (H9N2) the polysaccharase PB2 of subunit (PB2) gene, the part encoding sequence.

Gi|27466045|AY180717| influenza A virus strain A/ duck/Nanchang/1-0070/2000 (H9N2) the polysaccharase PB2 of subunit (PB2) gene, the part encoding sequence.

Gi|27466055|AY180722| influenza A virus strain A/ duck/Nanchang/10-389/2000 (H9N2) the polysaccharase PB2 of subunit (PB2) gene, the part encoding sequence.

Gi|27466057|AY180723| influenza A virus strain A/ pigeon/Nanchang/7-058/2000 (H9N2) the polysaccharase PB2 of subunit (PB2) gene, the part encoding sequence.

Gi|27466067|AY180728| influenza A virus strain A/ quail/Nanchang/2-0460/2000 (H9N2) the polysaccharase PB2 of subunit (PB2) gene, the part encoding sequence.

Gi|27466085|AY180737| influenza A virus strain A/ pigeon/Nanchang/11-145/2000 (H9N2) the polysaccharase PB2 of subunit (PB2) gene, the part encoding sequence.

Gi|27466087|AY180738| influenza A virus strain A/ duck/Nanchang/11-197/2000 (H9N2) the polysaccharase PB2 of subunit (PB2) gene, the part encoding sequence.

Gi|27466091|AY180740| influenza A virus strain A/ duck/Nanchang/11-290/2000 (H9N2) the polysaccharase PB2 of subunit (PB2) gene, the part encoding sequence.

Gi|27466093|AY180741| influenza A virus strain A/ duck/Nanchang/11-392/2000 (H9N2) the polysaccharase PB2 of subunit (PB2) gene, the part encoding sequence.

Gi|31339575|AF523463| influenza A virus (A/ duck/Shantou/2134/00 (H9N2)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|31339579|AF523465| influenza A virus (A/ duck/Shantou/1043/00 (H9N2)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|31339581|AF523466| influenza A virus (A/ duck/Shantou/1042/00 (H9N2)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|31339593|AF523472| influenza A virus (A/ duck/Shantou/2102/00 (H9N2)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|31339595|AF523473| influenza A virus (A/ duck/Shantou/2144/00 (H9N2)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|31339597|AF523474| influenza A virus (A/ duck/Shantou/2143/00 (H9N2)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|33318138|AF508654| influenza A virus (A/ chicken/Henan/62/00 (H9N2)) fragment 1 polysaccharase PB2 (PB2) gene, the part encoding sequence.

Gi|33318130|AF508650| influenza A virus (A/ chicken/Guangdong/10/00 (H9N2)) fragment 1 polysaccharase PB2 (PB2) gene, the part encoding sequence.

Gi|27466121|AY180755| influenza A virus strain A/ duck/Nanchang/7-092/2000 (H9N2) the polysaccharase PB2 of subunit (PB2) gene, the part encoding sequence.

Gi|27466141|AY180765| influenza A virus strain A/ chicken/Nanchang/4-010/2000 (H9N2) the polysaccharase PB2 of subunit (PB2) gene, the part encoding sequence.

Gi|27466157|AY180773| influenza A virus strain A/ quail/Nanchang/4-040/2000 (H9N2) the polysaccharase PB2 of subunit (PB2) gene, the part encoding sequence.

Gi|27466159|AY180774| influenza A virus strain A/ chicken/Nanchang/1-0016/2000 (H9N2) the polysaccharase PB2 of subunit (PB2) gene, the part encoding sequence.

Gi|55469788|AY768575| influenza A virus (A/ chicken/Korea S/SNU0028/00 (H9N2)) polysaccharase alkalescence subunit 2 (PB2) gene, the part encoding sequence.

Gi|55469790|AY768576| influenza A virus (A/ chicken/Korea S/SNU0037/00 (H9N2)) polysaccharase alkalescence subunit 2 (PB2) gene, the part encoding sequence.

Gi|55469792|AY768577| influenza A virus (A/ chicken/Korea S/SNU0073/00 (H9N2)) polysaccharase alkalescence subunit 2 (PB2) gene, the part encoding sequence.

Gi|55469794|AY768578| influenza A virus (A/ chicken/Korea S/SNU0091/00 (H9N2)) polysaccharase alkalescence subunit 2 (PB2) gene, the part encoding sequence.

Gi|55469796|AY768579| influenza A virus (A/ chicken/Korea S/SNU0140/00 (H9N2)) polysaccharase alkalescence subunit 2 (PB2) gene, the part encoding sequence.

Gi|55469798|AY768580| influenza A virus (A/ chicken/Korea S/SNU0146/00 (H9N2)) polysaccharase alkalescence subunit 2 (PB2) gene, the part encoding sequence.

Gi|55469800|AY768581| influenza A virus (A/ chicken/Korea S/SNU1035C/00 (H9N2)) polysaccharase alkalescence subunit 2 (PB2) gene, the part encoding sequence.

Gi|27466143|AY180766| influenza A virus strain A/ chicken/Nanchang/4-301/2001 (H9N2) the polysaccharase PB2 of subunit (PB2) gene, the part encoding sequence.

Gi|31339599|AF523475| influenza A virus (A/ duck/Shantou/2088/01 (H9N2)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|31339591|AF523471| influenza A virus (A/ duck/Shantou/1605/01 (H9N2)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|31339577|AF523464| influenza A virus (A/ wild duck/Shantou/4808/01 (H9N2)) polysaccharase (PB2) gene, the part encoding sequence.

Gi|27466097|AY180743| influenza A virus strain A/ chicken/Nanchang/4-361/2001 (H9N2) the polysaccharase PB2 of subunit (PB2) gene, the part encoding sequence.

Gi|54398631|AY664792| influenza A virus (A/ chicken/Hong Kong/CSW153/03 (H9N2)) polysaccharase basic protein 2 (PB2) gene, the part encoding sequence.

Gi|54398633|AY664793| influenza A virus (A/ chicken/Hong Kong/AP45/03 (H9N2)) polysaccharase basic protein 2 (PB2) gene, the part encoding sequence.

Gi|54398635|AY664794| influenza A virus (A/ chicken/Hong Kong/BD90/03 (H9N2)) polysaccharase basic protein 2 (PB2) gene, the part encoding sequence.

Gi|54398637|AY664795| influenza A virus (A/ chicken/Hong Kong/CSW291/03 (H9N2)) non-functional polysaccharase basic protein 2 (PB2) gene, partial sequence.

Gi|54398638|AY664796| influenza A virus (A/ chicken/Hong Kong/CSW304/03 (H9N2)) non-functional polysaccharase basic protein 2 (PB2) gene, partial sequence.

Gi|54398639|AY664797| influenza A virus (A/ chicken/Hong Kong/FY23/03 (H9N2)) non-functional polysaccharase basic protein 2 (PB2) gene, partial sequence.

Gi|54398640|AY664798| influenza A virus (A/ vulture galeeny/Hong Kong/NT101/03 (H9N2)) polysaccharase basic protein 2 (PB2) gene, the part encoding sequence.

Gi|54398642|AY664799| influenza A virus (A/ chicken/Hong Kong/NT142/03 (H9N2)) polysaccharase basic protein 2 (PB2) gene, the part encoding sequence.

Gi|54398644|AY664800| influenza A virus (A/ chicken/Hong Kong/SF1/03 (H9N2)) non-functional polysaccharase basic protein 2 (PB2) gene, partial sequence.

Gi|54398645|AY664801| influenza A virus (A/ chicken/Hong Kong/SSP101/03 (H9N2)) non-functional polysaccharase basic protein 2 (PB2) gene, partial sequence.

Gi|54398646|AY664802| influenza A virus (A/ chicken/Hong Kong/TP38/03 (H9N2)) polysaccharase basic protein 2 (PB2) gene, the part encoding sequence.

Gi|54398648|AY664803| influenza A virus (A/ chicken/Hong Kong/WF126/03 (H9N2)) non-functional polysaccharase basic protein 2 (PB2) gene, partial sequence.

Gi|54398649|AY664804| influenza A virus (A/ pigeon/Hong Kong/WF53/03 (H9N2)) non-functional polysaccharase basic protein 2 (PB2) gene, partial sequence.

Gi|54398650|AY664805| influenza A virus (A/ pheasant/Hong Kong/WF54/03 (H9N2)) non-functional polysaccharase basic protein 2 (PB2) gene, partial sequence.

Gi|54398651|AY664806| influenza A virus (A/ vulture galeeny/Hong Kong/NT184/03 (H9N2)) polysaccharase basic protein 2 (PB2) gene, the part encoding sequence.

Gi|54398653|AY664807| influenza A virus (A/ chicken/Hong Kong/WF120/03 (H9N2)) non-functional polysaccharase basic protein 2 (PB2) gene, partial sequence.

Gi|54398654|AY664808| influenza A virus (A/ chicken/Hong Kong/NT366/03 (H9N2)) polysaccharase basic protein 2 (PB2) gene, the part encoding sequence.

Gi|54398656|AY664809| influenza A virus (A/ chicken/Hong Kong/SSP418/03 (H9N2)) polysaccharase basic protein 2 (PB2) gene, the part encoding sequence.

Gi|54398658|AY664810| influenza A virus (A/ chicken/Hong Kong/YU427/03 (H9N2)) polysaccharase basic protein 2 (PB2) gene, the part encoding sequence.

Gi|55793686|AY800240| influenza A virus (A/ chicken/Korea S/S1/2003 (H9N2)) polysaccharase basic protein 2 (PB2) gene, the part encoding sequence.

Gi|58429686|AY862710| influenza A virus (A/ Gallus Domesticus/Korea S/S3/03 (H9N2)) PB2 (PB2) gene, the part encoding sequence.

Gi|58429688|AY862711| influenza A virus (A/ chicken/Korea S/S4/03 (H9N2)) PB2 (PB2) gene, the part encoding sequence.

Gi|58429690|AY862712| influenza A virus (A/ chicken/Korea S/S5/03 (H9N2)) PB2 (PB2) gene, the part encoding sequence.

Gi|58429692|AY862713| influenza A virus (A/ chicken/Korea S/S12/03 (H9N2)) PB2 (PB2) gene, the part encoding sequence.

Gi|58429694|AY862714| influenza A virus (A/ duck/Korea S/S13/03 (H9N2)) PB2 (PB2) gene, the part encoding sequence.

Gi|58429696|AY862715| influenza A virus (A/ dove/Korea S/S14/03 (H9N2)) PB2 (PB2) gene, the part encoding sequence.

Gi|58429698|AY862716| influenza A virus (A/ chicken/Korea S/S15/03 (H9N2)) PB2 (PB2) gene, the part encoding sequence.

Gi|58429700|AY862717| influenza A virus (A/ chicken/Korea S/S16/03 (H9N2)) PB2 (PB2) gene, the part encoding sequence.

Gi|58429702|AY862718| influenza A virus (A/ chicken/Korea S/S18/03 (H9N2)) PB2 (PB2) gene, the part encoding sequence.

Used sequence during influenza A polysaccharase acidic protein (PA) is analyzed

Gi|27465935|AY180662| influenza A virus strain A/ quail/Nanchang/12-340/2000 (H1N1) the polysaccharase PA of subunit (PA) gene, the part encoding sequence.

Gi|49357063|AY633217| influenza A virus (A/ wild duck/alberta is economized/211/98 (H1N1)) polymerase protein A (PA) gene, the part encoding sequence.

Gi|5918195|AJ243994| influenza A virus (strain A/ wild duck/New York/6750/78) the proteic part mRNA. of PA

Gi|45272157|AY422034| influenza A virus (A/ duck/Hokkaido/95/01 (H2N2)) PA albumen (PA) gene, the part encoding sequence.

Gi|27465965|AY180677| influenza A virus strain A/ chicken/Nanchang/3-120/2001 (H3N2) the polysaccharase PA of subunit (PA) gene, the part encoding sequence.

Gi|56159994|AY779263| influenza A virus (A/ turkey/North Carolina state/12344/03 (H3N2)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|56159996|AY779264| influenza A virus (A/ turkey/Minnesota State/764-2/03 (H3N2)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|58429736|AY862687| influenza A virus (A/ chicken/Korea S/S6/03 (H3N2)) PA (PA) gene, the part encoding sequence.

Gi|58429738|AY862688| influenza A virus (A/ duck/Korea S/S7/03 (H3N2)) PA (PA) gene, the part encoding sequence.

Gi|58429740|AY862689| influenza A virus (A/ duck/Korea S/S8/03 (H3N2)) PA (PA) gene, the part encoding sequence.

Gi|58429742|AY862690| influenza A virus (A/ duck/Korea S/S9/03 (H3N2)) PA (PA) gene, the part encoding sequence.

Gi|58429744|AY862691| influenza A virus (A/ duck/Korea S/S10/03 (H3N2)) PA (PA) gene, the part encoding sequence.

Gi|58429746|AY862692| influenza A virus (A/ dove/Korea S/S11/03 (H3N2)) PA (PA) gene, the part encoding sequence.

Gi|18091833|AF213914| influenza A virus (A/ chicken/Italy/5945/95 (H3N2)) fragment 3PA polymerase protein gene, the part encoding sequence.

Gi|58531086|AB166861| influenza A virus (A/ chicken/mountain pass/7/2004 (H5N1)) polysaccharase acidic protein PA gene, encoding sequence completely.

Gi|58531118|AB188815| influenza A virus (A/ chicken/big/8/2004 (H5N1) that divide) polysaccharase acidic protein PA gene, encoding sequence completely.

Gi|9863935|AF216739| influenza A virus (A/ environment/Hong Kong/437-10/99 (H5N1)) polysaccharase acidic protein gene, encoding sequence completely.

Gi|14165201|AF380163| influenza A virus (A/ goose/Guangdong/3/97 (H5N1)) fragment 3 polysaccharases (PA) gene, encoding sequence completely.

Gi|18092185|AF398427| influenza A virus (A/ goose/Hong Kong/385.3/2000 (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|18092187|AF398428| influenza A virus (A/ goose/Hong Kong/385.5/2000 (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|21359667|AF468841| influenza A virus (A/ duck/Anyang/AVL-1/2001 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|28849710|AF509195| influenza A virus (A/ chicken/Hong Kong/FY77/01 (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|28849712|AF509196| influenza A virus (A/ chicken/Hong Kong/YU562/01 (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|28849714|AF509197| influenza A virus (A/ chicken/Hong Kong/YU563/01 (H5N1)) polysaccharase (PA) gene, encoding sequence completely.

Gi|28849716|AF509198| influenza A virus (A/ chicken/Hong Kong/FY150/01 (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|28849718|AF509199| influenza A virus (A/ pheasant/Hong Kong/FY155/01 (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|28849720|AF509200| influenza A virus (A/ Gallus Domesticus/Hong Kong/SF189/01 (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|28849722|AF509201| influenza A virus (A/ quail/Hong Kong/SF203/01 (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|28849724|AF509202| influenza A virus (A/ pigeon/Hong Kong/SF215/01 (H5N1)) polymerase (PA) gene, the part encoding sequence.

Gi|28849726|AF509203| influenza A virus (A/ chicken/Hong Kong/SF219/01 (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|28849728|AF509204| influenza A virus (A/ chicken/Hong Kong/715.5/01 (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|28849730|AF509205| influenza A virus (A/ chicken/Hong Kong/751.1/01 (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|28849732|AF509206| influenza A virus (A/ chicken/Hong Kong/822.1/01 (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|28849734|AF509207| influenza A virus (A/ chicken/Hong Kong/829.2/01 (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|28849736|AF509208| influenza A virus (A/ chicken/Hong Kong/830.2/01 (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|28849738|AF509209| influenza A virus (A/ chicken/Hong Kong/858.3/01 (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|28849740|AF509210| influenza A virus (A/ chicken/Hong Kong/866.3/01 (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|28849742|AF509211| influenza A virus (A/ chicken/Hong Kong/867.1/01 (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|28849744|AF509212| influenza A virus (A/ chicken/Hong Kong/879.1/01 (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|28849746|AF509213| influenza A virus (A/ chicken/Hong Kong/873.3/01 (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|28849748|AF509214| influenza A virus (A/ chicken/Hong Kong/876.1/01 (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|28849750|AF509215| influenza A virus (A/ chicken/Hong Kong/891.1/01 (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|28849752|AF509216| influenza A virus (A/ chicken/Hong Kong/893.1/01 (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|28849754|AF509217| influenza A virus (A/ goose/Hong Kong/76.1/01 (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|28849756|AF509218| influenza A virus (A/ goose/Hong Kong/ww100/01 (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|28849758|AF509219| influenza A virus (A/ duck/Hong Kong/573.4/01 (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|28849760|AF509220| influenza A virus (A/ duck/Hong Kong/646.3/01 (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|19697861|AY059526| influenza A virus (A/ goose/Hong Kong/ww26/2000 (H5N1)) fragment 3 polysaccharases (PA) gene, the part encoding sequence.

Gi|19697863|AY059527| influenza A virus (A/ goose/Hong Kong/ww28/2000 (H5N1)) fragment 3 polysaccharases (PA) gene, the part encoding sequence.

Gi|19697865|AY059528| influenza A virus (A/ duck/Hong Kong/ww381/2000 (H5N1)) fragment 3 polysaccharases (PA) gene, the part encoding sequence.

Gi|19697869|AY059530| influenza A virus (A/ duck/Hong Kong/ww461/2000 (H5N1)) fragment 3 polysaccharases (PA) gene, the part encoding sequence.

Gi|19697871|AY059531| influenza A virus (A/ goose/Hong Kong/ww491/2000 (H5N1)) fragment 3 polysaccharases (PA) gene, the part encoding sequence.

Gi|19697873|AY059532| influenza A virus (A/ duck/Hong Kong/2986.1/2000 (H5N1)) fragment 3 polysaccharases (PA) gene, the part encoding sequence.

Gi|19697875|AY059533| influenza A virus (A/ goose/Hong Kong/3014.8/2000 (H5N1)) fragment 3 polysaccharases (PA) gene, the part encoding sequence.

Gi|28821204|AY221566| influenza A virus (A/ chicken/Hong Kong/NT873.3/01-MB (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|28821206|AY221567| influenza A virus (A/ chicken/Hong Kong/NT873.3/01 (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|28821208|AY221568| influenza A virus (A/ chicken/Hong Kong/FY150/01-MB (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|28821210|AY221569| influenza A virus (A/ chicken/Hong Kong/FY150/01 (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|28821212|AY221570| influenza A virus (A/ pheasant/Hong Kong/FY155/01-MB (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|28821214|AY221571| influenza A virus (A/ pheasant/Hong Kong/FY155/01 (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|28821216|AY221572| influenza A virus (A/ chicken/Hong Kong/YU822.2/01-MB (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|28821218|AY221573| influenza A virus (A/ chicken/Hong Kong/YU822.2/01 (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|28821220|AY221574| influenza A virus (A/ chicken/Hong Kong/YU562/01 (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|41207483|AY518365| influenza A virus (A/ duck/China/E319-2/03 (H5N1)) polysaccharase (PA) gene, encoding sequence completely.

Gi|51094114|AY551934| influenza A virus (A/ chicken/Buddhist system/Thailand/CU-K2/04 (H5N1)) polysaccharase (PA) gene, encoding sequence completely

Gi|47834839|AY576406| influenza A virus (A/Gs/ Hong Kong/739.2/02 (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|47834841|AY576407| influenza A virus (A/Eg/ Hong Kong/757.3/02 (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|47834843|AY576408| influenza A virus (A/G.H/ Hong Kong/793.1/02 (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|47834845|AY576409| influenza A virus (A/Dk/ Hong Kong/821/02 (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|47834847|AY576410| influenza A virus (A/Ck/ Hong Kong/31.4/02 (H5N1)) polysaccharase (PA) gene, encoding sequence completely.

Gi|47834849|AY576411| influenza A virus (A/Ck/ Hong Kong/61.9/02 (H5N1)) polysaccharase (PA) gene, encoding sequence completely.

Gi|47834851|AY576412| influenza A virus (A/Ck/ Hong Kong/YU777/02 (H5N1)) polysaccharase (PA) gene, encoding sequence completely.

Gi|47834853|AY576413| influenza A virus (A/Ck/ Hong Kong/96.1/02 (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|47834855|AY576414| influenza A virus (A/Ck/ Hong Kong/409.1/02 (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|47834857|AY576415| influenza A virus (A/Ph/ Hong Kong/sv674.15/02 (H5N1)) polysaccharase (PA) gene, encoding sequence completely.

Gi|47156478|AY585462| influenza A virus (A/ duck/Fujian/01/2002 (H5N1)) polysaccharase (PA) mRNA, encoding sequence completely.

Gi|47156480|AY585463| influenza A virus (A/ duck/Fujian/13/2002 (H5N1)) polysaccharase (PA) mRNA, encoding sequence completely.

Gi|47156482|AY585464| influenza A virus (A/ duck/Fujian/17/2001 (H5N1)) polysaccharase (PA) mRNA, encoding sequence completely.

Gi|47156484|AY585465| influenza A virus (A/ duck/Fujian/19/2000 (H5N1)) polysaccharase (PA) mRNA, encoding sequence completely.

Gi|47156486|AY585466| influenza A virus (A/ duck/Guangdong/01/2001 (H5N1)) polysaccharase (PA) mRNA, encoding sequence completely.

Gi|47156488|AY585467| influenza A virus (A/ duck/Guangdong/07/2000 (H5N1)) polysaccharase (PA) mRNA, encoding sequence completely.

Gi|47156490|AY585468| influenza A virus (A/ duck/Guangdong/12/2000 (H5N1)) polysaccharase (PA) mRNA, encoding sequence completely.

Gi|47156492|AY585469| influenza A virus (A/ duck/Guangdong/22/2002 (H5N1)) polysaccharase (PA) mRNA, encoding sequence completely.

Gi|47156494|AY585470| influenza A virus (A/ duck/Guangdong/40/2000 (H5N1)) polysaccharase (PA) mRNA, encoding sequence completely.

Gi|47156496|AY585471| influenza A virus (A/ duck/Guangxi/07/1999 (H5N1)) polysaccharase (PA) mRNA, encoding sequence completely.

Gi|47156498|AY585472| influenza A virus (A/ duck/Guangxi/22/2001 (H5N1)) polysaccharase (PA) mRNA, encoding sequence completely.

Gi|47156500|AY585473| influenza A virus (A/ duck/Guangxi/35/2001 (H5N1)) polysaccharase (PA) mRNA, encoding sequence completely.

Gi|47156502|AY585474| influenza A virus (A/ duck/Guangxi/50/2001 (H5N1)) polysaccharase (PA) mRNA, encoding sequence completely.

Gi|47156504|AY585475| influenza A virus (A/ duck/Guangxi/53/2002 (H5N1)) polysaccharase (PA) mRNA, encoding sequence completely.

Gi|47156506|AY585476| influenza A virus (A/ duck/Shanghai/08/2001 (H5N1)) polysaccharase (PA) mRNA, encoding sequence completely.

Gi|47156508|AY585477| influenza A virus (A/ duck/Shanghai/13/2001 (H5N1)) polysaccharase (PA) mRNA, encoding sequence completely.

Gi|47156510|AY585478| influenza A virus (A/ duck/Shanghai/35/2002 (H5N1)) polysaccharase (PA) mRNA, encoding sequence completely.

Gi|47156512|AY585479| influenza A virus (A/ duck/Shanghai/37/2002 (H5N1)) polysaccharase (PA) mRNA, encoding sequence completely.

Gi|47156514|AY585480| influenza A virus (A/ duck/Shanghai/38/2001 (H5N1)) polysaccharase (PA) mRNA, encoding sequence completely.

Gi|47156516|AY585481| influenza A virus (A/ duck/Zhejiang/11/2000 (H5N1)) polysaccharase (PA) mRNA, encoding sequence completely.

Gi|47156518|AY585482| influenza A virus (A/ duck/Zhejiang/52/2000 (H5N1)) polysaccharase (PA) mRNA, encoding sequence completely.

Gi|47716770|AY609311| influenza A virus (A/ chicken/Guangdong/174/04 (H5N1)) fragment 3, sequence completely.

Gi|50313026|AY651597| influenza A virus (A/Ck/ Indonesia/4/2004 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313028|AY651598| influenza A virus (A/Ck/ Indonesia/5/2004 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313030|AY651599| influenza A virus (A/Ck/ Indonesia/2A/2003 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313032|AY651600| influenza A virus (A/Dk/ Indonesia/MS/2004 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313034|AY651601| influenza A virus (A/Ck/ Indonesia/BL/2003 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313036|AY651602| influenza A virus (A/Ck/ Indonesia/PA/2003 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313038|AY651603| influenza A virus (A/Ck/ Thailand/1/2004 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313040|AY651604| influenza A virus (A/Ck/ Thailand/73/2004 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313042|AY651605| influenza A virus (A/Ck/ Thailand/9.1/2004 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313044|AY651606| influenza A virus (A/Qa/ Thailand/57/2004 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313046|AY651607| influenza A virus (A/bird/ Thailand/3.1/2004 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313048|AY651608| influenza A virus (A/Dk/ Thailand/71.1/2004 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313050|AY651609| influenza A virus (A/Gs/ Thailand/79/2004 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313060|AY651614| influenza A virus (A/Ck/ Vietnam/33/2004 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313062|AY651615| influenza A virus (A/Ck/ Vietnam/35/2004 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313064|AY651616| influenza A virus (A/Ck/ Vietnam/36/2004 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313066|AY651617| influenza A virus (A/Ck/ Vietnam/37/2004 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313068|AY651618| influenza A virus (A/Ck/ Vietnam/38/2004 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313070|AY651619| influenza A virus (A/Ck/ Vietnam/39/2004 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313072|AY651620| influenza A virus (A/Ck/ Vietnam/C57/2004 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313074|AY651621| influenza A virus (A/Dk/ Vietnam/11/2004 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313076|AY651622| influenza A virus (A/Gf/ Hong Kong/38/2002 (H5N1)) polysaccharase acidic protein (PA) gene, encoding sequence completely.

Gi|50313078|AY651623| influenza A virus (A/Ck/ Hong Kong/31.2/2002 (H5N1)) polysaccharase acidic protein (PA) gene, encoding sequence completely.

Gi|50313080|AY651624| influenza A virus (A/Ck/ Hong Kong/37.4/2002 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313082|AY651625| influenza A virus (A/SCk/ Hong Kong/YU100/2002 (H5N1)) polysaccharase acidic protein (PA) gene, encoding sequence completely.

Gi|50313084|AY651626| influenza A virus (A/Ck/ Hong Kong/YU22/2002 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313086|AY651627| influenza A virus (A/Ck/ Hong Kong/3176.3/2002 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313088|AY651628| influenza A virus (A/Ck/ Hong Kong/3169.1/2002 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313090|AY651629| influenza A virus (A/Ck/ Hong Kong/FY157/2003 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313092|AY651630| influenza A virus (A/Ck/ Hong Kong/YU324/2003 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313094|AY651631| influenza A virus (A/Ck/ Hong Kong/2133.1/2003 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313096|AY651632| influenza A virus (A/Ck/ Hong Kong/NT93/2003 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313098|AY651633| influenza A virus (A/Ck/ Hong Kong/SSP141/2003 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313100|AY651634| influenza A virus (A/Ck/ Hong Kong/WF157/2003 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313102|AY651635| influenza A virus (the red mouth of A/ gull/Hong Kong/12.1/2003 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313104|AY651636| influenza A virus (A/ heron/Hong Kong/861.1/2002 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313106|AY651637| influenza A virus (A/ rock dove/Hong Kong/862.7/2002 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313108|AY651638| influenza A virus (A/ sets sparrow/Hong Kong/864/2002 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313110|AY651639| influenza A virus (A/ teal/China/2978.1/2002 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313112|AY651640| influenza A virus (A/ falcon/Hong Kong/D0028/2004 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313114|AY651641| influenza A virus (A/Dk/HN/5806/2003 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313116|AY651642| influenza A virus (A/Dk/HN/303/2004 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313118|AY651643| influenza A virus (A/Dk/HN/101/2004 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313120|AY651644| influenza A virus (A/Dk/ST/4003/2003 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313122|AY651645| influenza A virus (A/Ph/ST/44/2004 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313124|AY651646| influenza A virus (A/Ck/ST/4231/2003 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313126|AY651647| influenza A virus (A/Dk/YN/6255/2003 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313128|AY651648| influenza A virus (A/Dk/YN/6445/2003 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313130|AY651649| influenza A virus (A/Ck/YN/115/2004 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50313132|AY651650| influenza A virus (A/Ck/YN/374/2004 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|50365724|AY653198| influenza A virus (A/ chicken/Jilin/9/2004 (H5N1)) fragment 3, sequence completely.

Gi|56548923|AY676029| influenza A virus (A/ duck/Hong Kong/821/02 (H5N1)) polysaccharase (PA) gene, encoding sequence completely.

Gi|56548925|AY676030| influenza A virus (A/ egression/Hong Kong/757.2/03 (H5N1)) polysaccharase (PA) gene, encoding sequence completely.

Gi|56548927|AY676031| influenza A virus (A/ chicken/Korea S/ES/03 (H5N1)) polysaccharase (PA) gene, encoding sequence completely.

Gi|56548929|AY676032| influenza A virus (A/ duck/Korea S/ESDI/03 (H5N1)) polysaccharase (PA) gene, encoding sequence completely.

Gi|50956625|AY684705| influenza A virus (A/ chicken/Hubei/327/2004 (H5N1)) polysaccharase (PA) gene, encoding sequence completely.

Gi|56119221|AY720944| influenza A virus (A/ chicken/Vietnam/DT-171/2004 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|56119227|AY720947| influenza A virus (A/ duck/Vietnam/TG-007A/2004 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|57924419|AY724784| influenza A virus (A/ chicken/Vietnam/HCM-022/2004 (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|57924480|AY724786| influenza A virus (A/ chicken/Vietnam/DN-045/2004 (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|57924569|AY724788| influenza A virus (A/ chicken/Vietnam/VL-008/2004 (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|57924680|AY724790| influenza A virus (A/ chicken/Vietnam/AG-010/2004 (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|57924765|AY724792| influenza A virus (A/ chicken/Vietnam/DT-015/2004 (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|57924882|AY724796| influenza A virus (A/ chicken/Vietnam/LA-024/2004 (H5N1)) polysaccharase (PA) gene, the part encoding sequence.

Gi|57915971|AY737288| influenza A virus (A/ chicken/Guangdong/191/04 (H5N1)) fragment 3, sequence completely.

Gi|57916018|AY737295| influenza A virus (A/ chicken/Guangdong/178/04 (H5N1)) fragment 3, sequence completely.

Gi|57916074|AY737303| influenza A virus (A/ duck/Guangdong/173/04 (H5N1)) fragment 3, sequence completely.

Gi|55233234|AY770082| influenza A virus (A/ chicken/Hubei/489/2004 (H5N1)) polymerase (PA) gene, encoding sequence completely.

Gi|54873465|AY770995| influenza A virus (A/ chicken/big subtlety/Thailand/CU-23/04 (H5N1)) pol gene, the part encoding sequence.

Gi|58618433|AY818133| influenza A virus (A/ chicken/Vietnam/C58/04 (H5N1)) polymerase protein PA gene, encoding sequence completely.

Gi|58618435|AY818134| influenza A virus (A/ quail/Vietnam/36/04 (H5N1)) polymerase protein PA gene, encoding sequence completely.

Gi|58374187|AY856863| influenza A virus (A/ duck/Shandong/093/2004 (H5N1)) fragment 3, sequence completely.

Gi|58531136|AB188823| influenza A virus (A/ chicken/capital of a country/3/2004 (H5N1)) polysaccharase acidic protein PA gene, encoding sequence completely.

Gi|58531154|AB189052| influenza A virus (A/ crow/capital of a country/53/2004 (H5N1)) polysaccharase acidic protein PA gene, encoding sequence completely.

Gi|58531170|AB189059| influenza A virus (A/ crow/Osaka/102/2004 (H5N1)) polysaccharase acidic protein PA gene, encoding sequence completely.

Gi|3335418|AF046087| influenza A virus (A/ chicken/Hong Kong/220/97 (H5N1)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|6048895|AF098604| influenza A virus (A/ chicken/Hong Kong/258/97 (H5N1)) PA albumen (PA) gene, encoding sequence completely.

Gi|6048897|AF098605| influenza A virus (A/ chicken/Hong Kong/y388/97 (H5N1)) PA albumen (PA) gene, encoding sequence completely.

Gi|6048899|AF098606| influenza A virus (A/ chicken/Hong Kong/728/97 (H5N1)) PA albumen (PA) gene, encoding sequence completely.

Gi|6048901|AF098607| influenza A virus (A/ chicken/Hong Kong/786/97 (H5N1)) PA albumen (PA) gene, encoding sequence completely.

Gi|6048903|AF098608| influenza A virus (A/ chicken/Hong Kong/915/97 (H5N1)) PA albumen (PA) gene, encoding sequence completely.

Gi|6048905|AF098609| influenza A virus (A/ duck/Hong Kong/p46/97 (H5N1)) PA albumen (PA) gene, encoding sequence completely.

Gi|6048907|AF098610| influenza A virus (A/ duck/Hong Kong/y283/97 (H5N1)) PA albumen (PA) gene, encoding sequence completely.

Gi|6048909|AF098611| influenza A virus (A/ goose/Hong Kong/w355/97 (H5N1)) PA albumen (PA) gene, encoding sequence completely.

Gi|5805280|AF144302| influenza A virus (A/ goose/Guangdong/1/96 (H5N1)) polysaccharase (PA) gene, encoding sequence completely.

Gi|9863880|AF216715| influenza A virus (A/ environment/Hong Kong/437-4/99 (H5N1)) polysaccharase acidic protein gene, encoding sequence completely.

Gi|9863899|AF216723| influenza A virus (A/ environment/Hong Kong/437-6/99 (H5N1)) polysaccharase acidic protein gene, encoding sequence completely.

Gi|9863917|AF216731| influenza A virus (A/ environment/Hong Kong/437-8/99 (H5N1)) polysaccharase acidic protein gene, encoding sequence completely.

Gi|34597776|AY303660| influenza A virus (A/ chicken/Chile/176822/02 (H7N3)) polysaccharase acidic protein gene, encoding sequence completely.

Gi|34597778|AY303661| influenza A virus (A/ chicken/Chile/4957/02 (H7N3)) polysaccharase acidic protein gene, encoding sequence completely.

Gi|34597780|AY303662| influenza A virus (A/ chicken/Chile/4322/02 (H7N3)) polysaccharase acidic protein gene, the part encoding sequence.

Gi|47680912|AY586431| influenza A virus (A/ wild duck/Italy/43/01 (H7N3)) PA gene, the part encoding sequence.

Gi|47680914|AY586432| influenza A virus (A/ wild duck/Italy/33/01 (H7N3)) PA gene, the part encoding sequence.

Gi|47680916|AY586433| influenza A virus (A/ turkey/Italy/220158/2002 (H7N3)) PA gene, the part encoding sequence.

Gi|47680918|AY586434| influenza A virus (A/ turkey/Italy/214845/02 (H7N3)) PA gene, the part encoding sequence.

Gi|47834370|AY616764| influenza A virus (A/ chicken/British Columbia/04 (H7N3)) polysaccharase acidic protein 2 (PA) gene, encoding sequence completely.

Gi|50542647|AY646083| influenza A virus (A/ chicken/British Columbia/GSC_ people _ B/04 (H7N3)) polysaccharase acidic protein 2 (PA) gene, encoding sequence completely.

Gi|50083049|AY648292| influenza A virus (A/GSC_ chicken _ B/ British Columbia/04 (H7N3)) polysaccharase acidic protein 2 (PA) gene, encoding sequence completely.

Gi|50059192|AY650275| influenza A virus (A/GSC_ chicken/British Columbia/04 (H7N3)) polysaccharase acidic protein 2 (PA) gene, encoding sequence completely.

Gi|9988639|AF268109| influenza A virus (the red abdomen dunlin of A/ (Retknot)/Delaware/259/94 (H7N7)) polymerase protein PA gene, the part encoding sequence.

Gi|40353080|AJ619677| influenza A virus (A/ chicken/Germany/R28/03 (H7N7)) polysaccharase compound subunit (PA) PA gene, geneome RNA.

Gi|37813167|AY342415| influenza A virus (A/ Holland/124/03 (H7N7)) polymerase protein A gene, the part encoding sequence.

Gi|37813169|AY342416| influenza A virus (A/ Holland/126/03 (H7N7)) polymerase protein A gene, the part encoding sequence.

Gi|37813171|AY342417| influenza A virus (A/ Holland/127/03 (H7N7)) polymerase protein A gene, the part encoding sequence.

Gi|37813173|AY342418| influenza A virus (A/ Holland/219/03 (H7N7)) polymerase protein A gene, encoding sequence completely.

Gi|37813175|AY342419| influenza A virus (A/ Holland/033/03 (H7N7)) polymerase protein A gene, encoding sequence completely.

Gi|37813177|AY342420| influenza A virus (A/ chicken/Holland/1/03 (H7N7)) polymerase protein A gene, encoding sequence completely.

Gi|13383274|AB049157| influenza A virus (A/ parakeet/Chiba/1/97 (H9N2)) polysaccharase acidic protein PA gene, encoding sequence completely.

Gi|13383276|AB049158| influenza A virus (A/ parakeet/Narita Airport/92A/98 (H9N2)) polysaccharase acidic protein PA gene, encoding sequence completely.

Gi|33318154|AF508662| influenza A virus (A/ ostrich/South Africa/9508103/95 (H9N2)) fragment 3 polysaccharase PA (PA) gene, encoding sequences completely.

Gi|33318156|AF508663| influenza A virus (A/ chicken/Pakistan/4/99 (H9N2)) fragment 3 polysaccharase PA (PA) gene, encoding sequences completely.

Gi|33318158|AF508664| influenza A virus (A/ chicken/Pakistan/5/99 (H9N2)) fragment 3 polysaccharase PA (PA) genes, the part encoding sequence.

Gi|33318160|AF508665| influenza A virus (A/ chicken/Germany/R45/98 (H9N2)) fragment 3 polysaccharase PA (PA) gene, encoding sequences completely.

Gi|33318162|AF508666| influenza A virus (A/ duck/Germany/113/95 (H9N2)) fragment 3 polysaccharase PA (PA) gene, encoding sequences completely.

Gi|33318164|AF508667| influenza A virus (A/ chicken/Iran/11T/99 (H9N2)) fragment 3 polysaccharase PA (PA) genes, the part encoding sequence.

Gi|33318166|AF508668| influenza A virus (A/ chicken/Saudi Arabia/532/99 (H9N2)) fragment 3 polysaccharase PA (PA) gene, encoding sequences completely.

Gi|33318168|AF508669| influenza A virus (A/ pheasant/Ireland/PV18/97 (H9N2)) fragment 3 polysaccharase PA (PA) gene, encoding sequences completely.

Gi|33318170|AF508670| influenza A virus (A/ chicken/Korea S/99029/99 (H9N2)) fragment 3 polysaccharase PA (PA) gene, encoding sequences completely.

Gi|33318172|AF508671| influenza A virus (A/ chicken/Beijing/8/98 (H9N2)) fragment 3 polysaccharase PA (PA) gene, encoding sequences completely.

Gi|33318174|AF508672| influenza A virus (A/ chicken/Guangdong/10/00 (H9N2)) fragment 3 polysaccharase PA (PA) gene, encoding sequences completely.

Gi|33318176|AF508673| influenza A virus (A/ chicken/Guangdong/11/97 (H9N2)) fragment 3 polysaccharase PA (PA) gene, encoding sequences completely.

Gi|33318178|AF508674| influenza A virus (A/ chicken/Hebei/4/98 (H9N2)) fragment 3 polysaccharase PA (PA) gene, encoding sequences completely.

Gi|33318180|AF508675| influenza A virus (A/ chicken/Heilungkiang/10/97 (H9N2)) fragment 3 polysaccharase PA (PA) gene, encoding sequences completely.

Gi|33318182|AF508676| influenza A virus (A/ chicken/Henan/62/00 (H9N2)) fragment 3 polysaccharase PA (PA) genes, the part encoding sequence.

Gi|33318184|AF508677| influenza A virus (A/ chicken/Ningxia/5/99 (H9N2)) fragment 3 polysaccharase PA (PA) gene, encoding sequences completely.

Gi|33318186|AF508678| influenza A virus (A/ chicken/Sichuan/5/97 (H9N2)) fragment 3 polysaccharase PA (PA) gene, encoding sequences completely.

Gi|33318188|AF508679| influenza A virus (A/ chicken/Shandong/6/96 (H9N2)) fragment 3 polysaccharase PA (PA) genes, the part encoding sequence.

Gi|33318190|AF508680| influenza A virus (A/ chicken/Shijiazhuang/2/99 (H9N2)) fragment 3 polysaccharase PA (PA) genes, the part encoding sequence.

Gi|33318192|AF508681| influenza A virus (A/ chicken/Shenzhen/9/97 (H9N2)) fragment 3 polysaccharase PA (PA) gene, encoding sequences completely.

Gi|33318194|AF508682| influenza A virus (A/ duck/Nanjing/1/97 (H9N2)) fragment 3 polysaccharase PA (PA) gene, encoding sequences completely.

Gi|33318196|AF508683| influenza A virus (A/ quail/Shanghai/8/96 (H9N2)) fragment 3 polysaccharase PA (PA) genes, the part encoding sequence.

Gi|31339541|AF523446| influenza A virus (A/ duck/Shantou/1043/00 (H9N2)) polysaccharase (PA) gene, the part encoding sequence.

Gi|31339543|AF523447| influenza A virus (A/ duck/Shantou/1042/00 (H9N2)) polysaccharase (PA) gene, the part encoding sequence.

Gi|31339545|AF523448| influenza A virus (A/ duck/Shantou/2088/01 (H9N2)) polysaccharase (PA) gene, the part encoding sequence.

Gi|31339547|AF523449| influenza A virus (A/ duck/Shantou/830/00 (H9N2)) polysaccharase (PA) gene, the part encoding sequence.

Gi|31339549|AF523450| influenza A virus (A/ duck/Shantou/1796/00 (H9N2)) polysaccharase (PA) gene, the part encoding sequence.

Gi|31339551|AF523451| influenza A virus (A/ duck/Shantou/2143/00 (H9N2)) polysaccharase (PA) gene, the part encoding sequence.

Gi|31339553|AF523452| influenza A virus (A/ duck/Shantou/2134/00 (H9N2)) polysaccharase (PA) gene, the part encoding sequence.

Gi|31339555|AF523453| influenza A virus (A/ duck/Shantou/2144/00 (H9N2)) polysaccharase (PA) gene, the part encoding sequence.

Gi|31339557|AF523454| influenza A virus (A/ wild duck/Shantou/4808/01 (H9N2)) polysaccharase (PA) gene, the part encoding sequence.

Gi|31339559|AF523455| influenza A virus (A/ duck/Shantou/1881/00 (H9N2)) polysaccharase (PA) gene, the part encoding sequence.

Gi|31339561|AF523456| influenza A virus (A/ duck/Shantou/2102/00 (H9N2)) polysaccharase (PA) gene, the part encoding sequence.

Gi|31339563|AF523457| influenza A virus (A/ duck/Hong Kong/289/78 (H9N2)) polysaccharase (PA) gene, the part encoding sequence.

Gi|31339565|AF523458| influenza A virus (A/ duck/Hong Kong/610/79 (H9N2)) polysaccharase (PA) gene, the part encoding sequence.

Gi|31339567|AF523459| influenza A virus (A/ duck/Hong Kong/86/76 (H9N2)) polysaccharase (PA) gene, the part encoding sequence.

Gi|31339569|AF523460| influenza A virus (A/ duck/Hong Kong/366/78 (H9N2)) polysaccharase (PA) gene, the part encoding sequence.

Gi|22759040|AF536669| influenza A virus (A/ chicken/Beijing/1/95 (H9N2)) PA gene, the part encoding sequence.

Gi|22759042|AF536670| influenza A virus (A/ chicken/Beijing/2/97 (H9N2)) PA gene, the part encoding sequence.

Gi|22759044|AF536671| influenza A virus (A/ chicken/Beijing/3/99 (H9N2)) PA gene, the part encoding sequence.

Gi|22759046|AF536672| influenza A virus (A/ chicken/Guangdong/97 (H9N2)) PA gene, the part encoding sequence.

Gi|22759048|AF536673| influenza A virus (A/ chicken/Hebei/1/96 (H9N2)) PA gene, the part encoding sequence.

Gi|22759050|AF536674| influenza A virus (A/ chicken/Hebei/2/98 (H9N2)) PA gene, the part encoding sequence.

Gi|22759052|AF536675| influenza A virus (A/ chicken/Hebei/3/98 (H9N2)) PA gene, the part encoding sequence.

Gi|22759054|AF536676| influenza A virus (A/ chicken/Henan/98 (H9N2)) PA gene, the part encoding sequence.

Gi|22759056|AF536677| influenza A virus (A/ chicken/Liaoning/99 (H9N2)) PA gene, the part encoding sequence.

Gi|22759058|AF536678| influenza A virus (A/ chicken/Shandong/98 (H9N2)) PA gene, the part encoding sequence.

Gi|12038897|AJ291397| influenza A virus (A/ chicken/Pakistan/2/99 (H9N2)) polysaccharase (PA) PA gene, geneome RNA.

Gi|18496121|AJ427863| influenza A virus (A/ quail/Hong Kong/FY298/00 (H9N2)) PA polymerase protein part pa gene, geneome RNA

Gi|27465911|AY180650| influenza A virus strain A/ duck/Nanchang/11-392/2000 (H9N2) the polysaccharase PA of subunit (PA) gene, the part encoding sequence.

Gi|27465913|AY180651| influenza A virus strain A/ duck/Nanchang/11-290/2000 (H9N2) the polysaccharase PA of subunit (PA) gene, the part encoding sequence.

Gi|27465915|AY180652| influenza A virus strain A/ chicken/Nanchang/1-0016/2000 (H9N2) the polysaccharase PA of subunit (PA) gene, the part encoding sequence.

Gi|27465917|AY180653| influenza A virus strain A/ duck/Nanchang/11-197/2000 (H9N2) the polysaccharase PA of subunit (PA) gene, the part encoding sequence.

Gi|27465937|AY180663| influenza A virus strain A/ pigeon/Nanchang/2-0461/2000 (H9N2) the polysaccharase PA of subunit (PA) gene, the part encoding sequence.

Gi|27465941|AY180665| influenza A virus strain A/ chicken/Nanchang/4-301/2001 (H9N2) the polysaccharase PA of subunit (PA) gene, the part encoding sequence.

Gi|27465943|AY180666| influenza A virus strain A/ chicken/Nanchang/4-361/2001 (H9N2) the polysaccharase PA of subunit (PA) gene, the part encoding sequence.

Gi|27465961|AY180675| influenza A virus strain A/ wild duck/Nanchang/2-0480/2000 (H9N2) the polysaccharase PA of subunit (PA) gene, the part encoding sequence.

Gi|27465983|AY180686| influenza A virus strain A/ duck/Nanchang/1-0070/2000 (H9N2) the polysaccharase PA of subunit (PA) gene, the part encoding sequence.

Gi|27465989|AY180689| influenza A virus strain A/ duck/Nanchang/10-389/2000 (H9N2) the polysaccharase PA of subunit (PA) gene, the part encoding sequence.

Gi|27465993|AY180691| influenza A virus strain A/ pigeon/Nanchang/11-145/2000 (H9N2) the polysaccharase PA of subunit (PA) gene, the part encoding sequence.

Gi|27466001|AY180695| influenza A virus strain A/ quail/Nanchang/2-0460/2000 (H9N2) the polysaccharase PA of subunit (PA) gene, the part encoding sequence.

Gi|27466003|AY180696| influenza A virus strain A/ quail/Nanchang/4-040/2000 (H9N2) the polysaccharase PA of subunit (PA) gene, the part encoding sequence.

Gi|27466009|AY180699| influenza A virus strain A/ chicken/Nanchang/4-010/2000 (H9N2) the polysaccharase PA of subunit (PA) gene, the part encoding sequence.

Gi|27466015|AY180702| influenza A virus strain A/ duck/Nanchang/7-092/2000 (H9N2) the polysaccharase PA of subunit (PA) gene, the part encoding sequence.

Gi|27466019|AY180704| influenza A virus strain A/ pigeon/Nanchang/7-058/2000 (H9N2) the polysaccharase PA of subunit (PA) gene, the part encoding sequence.

Gi|30025973|AY253752| influenza A virus (A/ chicken/Shanghai/F/98 (H9N2)) polysaccharase acidic protein (PA) gene, encoding sequence completely.

Gi|49357051|AY633169| influenza A virus (A/ wild duck/alberta is economized/17/91 (H9N2)) polymerase protein A (PA) gene, the part encoding sequence.

Gi|49357079|AY633281| influenza A virus (A/ wild duck/alberta is economized/321/88 (H9N2)) polymerase protein A (PA) gene, the part encoding sequence.

Gi|49357083|AY633297| influenza A virus (A/ wild duck/alberta is economized/11/91 (H9N2)) polymerase protein A (PA) gene, the part encoding sequence.

Gi|54301528|AY664755| influenza A virus (A/ chicken/Hong Kong/CSW153/03 (H9N2)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|54301530|AY664756| influenza A virus (A/ chicken/Hong Kong/AP45/03 (H9N2)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|54301532|AY664757| influenza A virus (A/ chicken/Hong Kong/BD90/03 (H9N2)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|54301534|AY664758| influenza A virus (A/ chicken/Hong Kong/CSW291/03 (H9N2)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|54301536|AY664759| influenza A virus (A/ chicken/Hong Kong/CSW304/03 (H9N2)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|54301538|AY664760| influenza A virus (A/ chicken/Hong Kong/FY23/03 (H9N2)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|54301540|AY664761| influenza A virus (A/ vulture galeeny/Hong Kong/NT10I/03 (H9N2)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|54301542|AY664762| influenza A virus (A/ chicken/Hong Kong/NT142/03 (H9N2)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|54301544|AY664763| influenza A virus (A/ chicken/Hong Kong/SF1/03 (H9N2)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|54301546|AY664764| influenza A virus (A/ chicken/Hong Kong/SSP101/03 (H9N2)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|54301548|AY664765| influenza A virus (A/ chicken/Hong Kong/TP38/03 (H9N2)) polysaccharase acidic protein sample (PA) gene, sequence completely.

Gi|54301549|AY664766| influenza A virus (A/ chicken/Hong Kong/WF126/03 (H9N2)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|54301551|AY664767| influenza A virus (A/ pigeon/Hong Kong/WF53/03 (H9N2)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|54301553|AY664768| influenza A virus (A/ pheasant/Hong Kong/WF54/03 (H9N2)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|54301555|AY664769| influenza A virus (A/ vulture galeeny/Hong Kong/NT184/03 (H9N2)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|54301557|AY664770| influenza A virus (A/ chicken/Hong Kong/WF120/03 (H9N2)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|54301559|AY664771| influenza A virus (A/ chicken/Hong Kong/NT366/03 (H9N2)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|54301561|AY664772| influenza A virus (A/ chicken/Hong Kong/SSP418/03 (H9N2)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|54301563|AY664773| influenza A virus (A/ chicken/Hong Kong/YU427/03 (H9N2)) polysaccharase acidic protein (PA) gene, the part encoding sequence.

Gi|55793682|AY800238| influenza A virus (A/ chicken/Korea S/S1/2003 (H9N2)) polysaccharase acidic protein (PA) gene, encoding sequence completely.

Gi|58429718|AY862678| influenza A virus (A/silky chicken/Korea S/S3/03 (H9N2)) PA (PA) gene, the part encoding sequence.

Gi|58429720|AY862679| influenza A virus (A/ chicken/Korea S/S4/03 (H9N2)) PA (PA) gene, the part encoding sequence.

Gi|58429722|AY862680| influenza A virus (A/ chicken/Korea S/S5/03 (H9N2)) PA (PA) gene, encoding sequence completely.

Gi|58429724|AY862681| influenza A virus (A/ chicken/Korea S/S12/03 (H9N2)) PA (PA) gene, the part encoding sequence.

Gi|58429726|AY862682| influenza A virus (A/ duck/Korea S/S13/03 (H9N2)) PA (PA) gene, the part encoding sequence.

Gi|58429728|AY862683| influenza A virus (A/ dove/Korea S/S14/03 (H9N2)) PA (PA) gene, the part encoding sequence.

Gi|58429730|AY862684| influenza A virus (A/ chicken/Korea S/S15/03 (H9N2)) PA (PA) gene, the part encoding sequence.

Gi|58429732|AY862685| influenza A virus (A/ chicken/Korea S/S16/03 (H9N2)) PA (PA) gene, the part encoding sequence.

Gi|58429734|AY862686| influenza A virus (A/ chicken/Korea S/S18/03 (H9N2)) PA (PA) gene, the part encoding sequence.

Gi|5732356|AF156444| influenza A virus (A/ chicken/Hong Kong/G9/97 (H9N2)) fragment 3 polysaccharases (PA) gene, the part encoding sequence.

Gi|5732358|AF156445| influenza A virus (A/ chicken/Hong Kong/G23/97 (H9N2)) fragment 3 polysaccharases (PA) gene, encoding sequence completely.

Gi|5732360|AF156446| influenza A virus (A/ pigeon/Hong Kong/Y233/97 (H9N2)) fragment 3 polysaccharases (PA) gene, the part encoding sequence.

Gi|5732362|AF156447| influenza A virus (A/ duck/Hong Kong/Y280/97 (H9N2)) fragment 3 polysaccharases (PA) gene, the part encoding sequence.

Gi|5732364|AF156448| influenza A virus (A/ duck/Hong Kong/Y439/97 (H9N2)) fragment 3 polysaccharases (PA) gene, the part encoding sequence.

Gi|5732366|AF156449| influenza A virus (A/ quail/Hong Kong/G1/97 (H9N2)) fragment 3 polysaccharases (PA) gene, the part encoding sequence.

Gi|5732368|AF156450| influenza A virus (A/ chicken/Hong Kong/739/94 (H9N2)) fragment 3 polysaccharases (PA) gene, the part encoding sequence.

Gi|5732370|AF156451| influenza A virus (A/ quail/Hong Kong/AF157/92 (H9N2)) fragment 3 polysaccharases (PA) gene, the part encoding sequence.

Gi|5732372|AF156452| influenza A virus (A/ chicken/Beijing/1/94 (H9N2)) fragment 3 polysaccharases (PA) gene, the part encoding sequence.

Gi|5732374|AF156453| influenza A virus (A/ chicken/Korea S/38349-p96323/96 (H9N2)) fragment 3 polysaccharases (PA) gene, the part encoding sequence.

Gi|5732376|AF156454| influenza A virus (A/ chicken/Korea S/25232-96006/96 (H9N2)) fragment 3 polysaccharases (PA) gene, the part encoding sequence.

Gi|5732378|AF156455| influenza A virus (A/Shorebird/ Delaware/9/96 (H9N2)) fragment 3 polysaccharases (PA) gene, the part encoding sequence.

Gi|5732380|AF156456| influenza A virus (A/ quail/Arkansas State/29209-1/93 (H9N2)) fragment 3 polysaccharases (PA) gene, the part encoding sequence.

Gi|5732382|AF156457| influenza A virus (A/ turkey/California/189/66 (H9N2)) fragment 3 polysaccharases (PA) gene, the part encoding sequence.

Gi|12060671|AF222642| influenza A virus (A/ quail/Hong Kong/A17/99 (H9N2)) fragment 3 PA (PA) gene, the part encoding sequence.

Gi|12060673|AF222643| influenza A virus (A/ pigeon/Hong Kong/FY6/99 (H9N2)) fragment 3 PA (PA) gene, the part encoding sequence.

Gi|12060675|AF222644| influenza A virus (A/ chicken/Hong Kong/NT16/99 (H9N2)) fragment 3 PA (PA) gene, the part encoding sequence.

Gi|12060677|AF222645| influenza A virus (A/ quail/Hong Kong/SSP10/99 (H9N2)) fragment 3 PA (PA) gene, the part encoding sequence.

Gi|12060679|AF222646| influenza A virus (A/ pheasant/Hong Kong/SSP11/99 (H9N2)) fragment 3 PA (PA) gene, the part encoding sequence.

Gi|12060681|AF222647| influenza A virus (A/ chicken/Hong Kong/FY20/99 (H9N2)) fragment 3 PA (PA) gene, the part encoding sequence.

Gi|12060683|AF222648| influenza A virus (A/ chicken/Hong Kong/KC12/99 (H9N2)) fragment 3 PA (PA) gene, the part encoding sequence.

Gi|12060685|AF222649| influenza A virus (A/ quail/Hong Kong/NT28/99 (H9N2)) fragment 3 PA (PA) gene, the part encoding sequence.

Gi|12060687|AF222650| influenza A virus (A/ chicken/Hong Kong/SF2/99 (H9N2)) fragment 3 PA (PA) gene, the part encoding sequence.

Gi|12060689|AF222651| influenza A virus (A/ Gallus Domesticus/Hong Kong/SF44/99 (H9N2)) fragment 3 PA (PA) gene, the part encoding sequence.

Claims (23)

1. the iRNA reagent that comprises sense strand and antisense strand, wherein said sense strand comprises at least 15 successive Nucleotide, described successive Nucleotide and the reagent that provides of table 1A-1H are numbered no more than 1,2 or 3 of the different IPs thuja acid of sense strand sequence of arbitrary reagent of AL-DP-2241-AL-DP-8631, wherein said antisense strand comprises at least 15 successive Nucleotide, and the reagent that described successive Nucleotide and table provide among the 1A-1H is numbered no more than 1,2 or 3 of the different IPs thuja acid of antisense sequences of arbitrary reagent of AL-DP-2241-AL-DP-8631.

2. the iRNA reagent that comprises sense strand and antisense strand, wherein said sense strand comprises at least 15 successive Nucleotide, described successive Nucleotide and the reagent that provides of table 1A-1H are numbered the different IPs thuja acid no more than 1 of sense strand sequence of arbitrary reagent of AL-DP-2241-AL-DP-8631,2 or 3, the reagent that wherein said antisense strand comprises table to be provided among the 1A-1H is numbered at least 15 successive Nucleotide of antisense sequences of arbitrary reagent of AL-DP-2241-AL-DP-8631, and described iRNA reagent makes the target gene expression of expressing in the Cos-7 cell of target gene separately through transformation separately reduce by 20% than the expression in the cell of iRNA reagent incubation not, 30%, 40%, 50%, 60%, more than 70% or 80%.

3. the iRNA reagent that comprises sense strand and antisense strand, sense strand and antisense strand all comprise at least 16,17 or 18 nucleotide sequences, to be numbered the sequence of arbitrary reagent of AL-DP-2241-AL-DP-8631 identical with reagent that table provides among the 1A-1H basically for described sequence, different is that each chain has no more than 1,2 or 3 Nucleotide to be replaced (for example adenosine is replaced by uridylic) by other Nucleotide, forms the ability that reduces the amount of the influenza A plaque that forms in the cell in the detection but still kept basically in plaque.

4. each described iRNA reagent in the claim 1-3, the length of wherein said sense-rna chain are 30 Nucleotide or still less, and the length in the duplex zone of described iRNA reagent is 15-30 nucleotide pairs.

5. each described iRNA reagent in the claim 1-3, it comprises can make the modification that stability increases in biological sample of described iRNA reagent.

6. each described iRNA reagent in the claim 1-3, it comprises thiophosphatephosphorothioate or the 2 ' Nucleotide of modifying.

7. each described iRNA reagent in the claim 1-3, it comprises at least one 5 '-uridine-VITAMIN B4-3 ' (5 '-ua-3 ') dinucleotides, and wherein said uridine is the 2 ' Nucleotide of modifying; At least one 5 '-uridine-guanine-3 ' (5 '-ug-3 ') dinucleotides, wherein said 5 '-uridine is the 2 ' Nucleotide of modifying; At least one 5 '-cytidine-VITAMIN B4-3 ' (5 '-ca-3 ') dinucleotides, wherein said 5 '-cytidine is the 2 ' Nucleotide of modifying; Or at least one 5 '-uridine-uridine-3 ' (5 '-uu-3 ') dinucleotides, wherein said 5 '-uridine is the 2 ' Nucleotide of modifying.

8. claim 6 or 7 described iRNA reagent, wherein said 2 ' modifies the group that is selected from following composition: 2 '-deoxidation, 2 '-deoxidation-2 '-fluorine, 2 '-O-methyl, 2 '-the O-methoxy ethyl (2 '-O-MOE), 2 '-the O-aminopropyl (2 '-O-AP), 2 '-the O-dimethyl aminoethyl (2 '-O-DMAOE), 2 '-the O-dimethylaminopropyl (2 '-O-DMAP), 2 '-O-dimethyl aminoethyl oxygen ethyl (2 '-O-DMAEOE) and 2 '-O-N-methyl kharophen (2 '-O-NMA).

9. each described iRNA reagent of claim 1-3, it comprises the Nucleotide overhang with 1-4 unpaired nucleotides.

10. the described iRNA reagent of claim 9, the overhang of wherein said Nucleotide has 2 or 3 unpaired Nucleotide.

11. the described iRNA reagent of claim 9, wherein said Nucleotide overhang is at 3 ' end of described iRNA reagent antisense strand.

12. each described iRNA reagent of claim 1-3, it comprises cholesterol moiety.

13. the described iRNA reagent of claim 12, wherein said cholesterol moiety combines with 3 ' end of described iRNA reagent sense strand.

14. each described iRNA reagent of claim 1-3, wherein said iRNA reagent target is in being absorbed by pneumonocyte.

15. each described iRNA reagent of claim 1-3, wherein said iRNA reagent comprises at least a non-natural base.

16. the described iRNA reagent of claim 15, wherein said non-natural base are difluoro toluene base, nitroindoline base, nitro-pyrrole base or nitroimidazole base.

17. the described iRNA reagent of claim 15, wherein said non-natural base is the difluoro toluene base.

18. the described iRNA reagent of claim 15 only wherein comprises in two oligonucleotide chains of double chain oligonucleotide one and contains the non-natural base.

19. the described iRNA reagent of claim 15, two oligonucleotide chains that wherein comprise double chain oligonucleotide contain the non-natural base respectively.

20. treat its pathologic process part is duplicated the experimenter who is mediated by influenza A virus method, wherein said iRNA pack contains sense strand and antisense strand, wherein said sense strand comprises at least 15 successive Nucleotide, described successive Nucleotide and the reagent that provides of table 1A-1H are numbered the different IPs thuja acid no more than 1 of sense strand sequence of arbitrary reagent of AL-DP-2241-AL-DP-8631,2 or 3, wherein said antisense strand comprises at least 15 successive Nucleotide, and the reagent that described successive Nucleotide and table provide among the 1A-1H is numbered the different IPs thuja acid no more than 1 of antisense strand sequence of arbitrary reagent of AL-DP-2241-AL-DP-8631,2 or 3.

21. method according to claim 20, wherein said iRNA reagent is used with the amount of duplicating that enough reduces influenza virus in experimenter's cell or tissue.

22. method according to claim 20, wherein said experimenter is the people.

23. pharmaceutical composition comprises:

A.) each described iRNA reagent in the claim 1-3;

B.) pharmaceutical acceptable carrier.

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