CN107586885A

CN107586885A - A kind of LAMP primer group and kit for being used to expand influenza A virus

Info

Publication number: CN107586885A
Application number: CN201711026762.9A
Authority: CN
Inventors: 杜昱光; 齐立斐; 毛瑞; 王倬
Original assignee: Institute of Process Engineering of CAS
Current assignee: Institute of Process Engineering of CAS
Priority date: 2017-10-27
Filing date: 2017-10-27
Publication date: 2018-01-16

Abstract

The present invention relates to biological technical field, more particularly to a kind of LAMP primer group and kit for being used to detect influenza A virus (Influenza A virus, IAV).The primer sets include the one or more in following 29 groups of primer sets：Primer sets for IAV H1~H18 areas fragment amplification and present invention also offers the kit for detecting IAV viruses for the IAV primer sets of N1~N11 areas fragment amplification.Primer sets sensitiveness provided by the invention is high, high specificity, and the kit being made from it can fast and accurately detect the IAV contained in testing sample, and can carry out parting detection and identification to the IAV types in testing sample.The present invention using artificial RNA as during control test, non-false positive and false negative phenomenon generation.Primer sensitivity and specificity provided by the invention are all very high, and the visualizing agent box provided will provide great convenience for Site Detection, are prepared into realize for kit and IAV is fast and accurately detected.

Description

A kind of LAMP primer group and kit for being used to expand influenza A virus

Technical field

The present invention relates to biological technical field, more particularly to one kind to be used to expand influenza A virus (Influenza A Virus, IAV) LAMP primer group and kit.

Background technology

Influenza virus belongs to orthomyxovirus section, can be divided into A type, B-mode and the third type.Wherein, A type and influenza B virus Caused seasonal influenza, morbidity and mortality are higher, constantly trigger global health problem, caused by economic loss it is huge Greatly.There are hemagglutinin (hemagglutinin, HA) and neuraminidase on Alphavirus particle surface cyst membrane (neuraminidase, NA) two kinds of surfaces are fine prominent, are two kinds of important glycoprotein, and the surface antigen that virus is main, according to HA is different from NA's to divide into different hypotypes, such as H1N1 and H7N9 etc., at present it has been found that H hypotypes have 18 kinds of (H1 ~H18), and N hypotypes have 11 kinds (N1~N11), the virus of these hypotypes nearly all can from aquatic birds, poultry and other Birds kind is separated to.Highly pathogenic influenza can not only cause huge economic losses to aviculture, while can also cross over host's screen Hinder direct infection people, serious threat public health security.The example being most widely known by the people is at present in several areas of Asia and Africa The popular subtype avian influenza virus H5N1 in the Northeast, it was reported that the virus caused the mankind ill since 1997 and death, Serious public health security problem is caused, therefore is listed in the highly pathogenic epidemic disease for needing to report, is also classified as one by China Class animal epidemic.

The vaccine of influenza A virus and medicine can only be directed to specific type at present, and there is no accurate A type so far The full genotyping detection method of influenza virus.Also because influenza A virus blood serum subtype is numerous, antigenic variability is strong, therefore for this The prevention and control of disease then seem particularly important.And quick, special, the highly sensitive diagnosis for the virus is then important in prevention and control One ring.IAV method for detecting virus is recommended to be directed to 2 sections and above genome spy based on RT-PCR by the World Health Organization (WHO) The amplification of signal detection of specific nucleic acid sequences, but application RT-PCR method in standard biochemical assay laboratory, it is necessary to use essence Close real-time fluorescence PCR instrument, and reagent preparation, sample preparation and three independent test blocks of PCR augmentation detections are needed, be not suitable for For on-site quick screening.And the test strips made by immunoassay technology have simple and efficient advantage, but this method flase drop Rate is higher, sensitivity is relatively low, it is difficult to which detection is in preclinical IAV carrier, may cause the diffusion of epidemic situation.Therefore, towards The quick, sensitive of scene, the research and development of high specific nucleic acids marker detection technique will provide strong guarantee for IAV prevention and control.

Loop-mediated isothermal amplification technique (loop-mediated isothermal amplification, LAMP) is 2000 The replacement PCR nucleic acid amplification methods of year Japanese Eiken Chemical's exploitation.This method is mainly different special using 4 kinds Property primer identification target gene specific region, isothermy carry out amplified reaction.Loop-mediated isothermal amplification technique (LAMP) is no The special instruments and equipment of costliness is needed, reaction result can use direct visual perception, be highly suitable for development at the basic level pathogenic microorganism Early diagnosis and screening.It is characterized under the conditions of constant temperature (about 65 DEG C), 4 is designed using six sections of regions for target sequence Special primer, can be by the RNA efficient amplifications of several copies to 10 in 1h⁹~10¹⁰Times.With conventional gene detection means (such as PCR Deng) compare, LAMP reactions can be completed in constant water bath box, and instrument and equipment requires low, simple to operate compared with normal PCR and cultivation It is many, it is not necessary to which that professional also can accurately complete, and be adapted to the application of basic medical unit.Also, LAMP can also contract significantly The short operation time, reduce sample opportunities for contamination, be suitably applied IAV quick diagnosis.

At present, the full parting detection techniques of LAMP on IAV more conserved positions have not been reported.

The content of the invention

Present invention mainly solves technical problem be to provide a kind of 29 sections of conserved sequences for being used to expand IAV LAMP primer group and kit and application, primer sensitivity and specificity provided by the invention are all very high, are prepared into as examination Agent box can realize that fast and accurately parting detects to IAV.

The LAMP primer group for being used to expand influenza A virus of the present invention, it is characterised in that the LAMP primer group bag Include the one or more in following 29 groups of primer sets：For IAV H1 fragments (such as GenBank:U47310.1, its core Nucleotide sequence is as shown in SEQ ID NO.117) primer sets of amplification, H2 fragments (such as the GenBank for IAV: AY207522.1, its nucleotide sequence is as shown in SEQ ID NO.118) amplification primer sets, for IAV H3 fragments (such as GenBank:V01087.1, its nucleotide sequence is as shown in SEQ ID NO.119) amplification primer sets, the H4 fragments for IAV (such as GenBank:D90302.1, its nucleotide sequence is as shown in SEQ ID NO.120) amplification primer sets, for IAV H5 fragments (such as GenBank:AF144305.1, its nucleotide sequence is as shown in SEQ ID NO.121) amplification primer sets, pin To IAV H6 fragments (such as GenBank:AY968676.1, its nucleotide sequence is as shown in SEQ ID NO.122) amplification draw Thing group, H7 fragments (such as the GenBank for IAV:AJ584647.1, its nucleotide sequence is as shown in SEQ ID NO.123) The primer sets of amplification, H8 fragments (such as the GenBank for IAV:D90304.1, its nucleotide sequence such as SEQ ID NO.124 It is shown) primer sets of amplification, H9 fragments (such as the GenBank for IAV:D90305.1, its nucleotide sequence such as SEQ ID Shown in NO.125) primer sets of amplification, H10 fragments (such as the GenBank for IAV:M21647.1, its nucleotide sequence is such as Shown in SEQ ID NO.126) primer sets of amplification, H11 fragments (such as the GenBank for IAV:D90306.1, its nucleotides Sequence is as shown in SEQ ID NO.127) primer sets of amplification, H12 fragments (such as the GenBank for IAV:D90307.1, its Nucleotide sequence is as shown in SEQ ID NO.128) primer sets of amplification, H13 fragments (such as the GenBank for IAV: M26090.1, its nucleotide sequence is as shown in SEQ ID NO.129) amplification primer sets, for IAV H14 fragments (such as GenBank:M35997.1, its nucleotide sequence is as shown in SEQ ID NO.130) amplification primer sets, the H15 pieces for IAV Section (such as GenBank:L43916.1, its nucleotide sequence is as shown in SEQ ID NO.131) amplification primer sets, for IAV H16 fragments (such as GenBank:CY177441.1, its nucleotide sequence is as shown in SEQ ID NO.132) amplification primer Group, H17 fragments (such as the GenBank for IAV:CY103892.1, its nucleotide sequence is as shown in SEQ ID NO.133) expand The primer sets of increasing, H18 fragments (such as the GenBank for IAV:CY125945.1, its nucleotide sequence such as SEQ ID Shown in NO.134) primer sets of amplification, N1 fragments (such as the GenBank for IAV:GQ290690.1, its nucleotide sequence is such as Shown in SEQ ID NO.135) primer sets of amplification, N2 fragments (such as the GenBank for IAV:KC190181.1, its nucleosides Acid sequence is as shown in SEQ ID NO.136) primer sets of amplification, N3 fragments (such as the GenBank for IAV: AY207524.1, its nucleotide sequence is as shown in SEQ ID NO.137) amplification primer sets, for IAV N4 fragments (such as GenBank:AY207531.1, its nucleotide sequence is as shown in SEQ ID NO.138) amplification primer sets, the N5 pieces for IAV Section (such as GenBank:KM244094.1, its nucleotide sequence is as shown in SEQ ID NO.139) amplification primer sets, be directed to IAV N6 fragments (such as GenBank:AY207556.1, its nucleotide sequence is as shown in SEQ ID NO.140) amplification primer Group, N7 fragments (such as the GenBank for IAV:KM244102.1, its nucleotide sequence is as shown in SEQ ID NO.141) expand The primer sets of increasing, N8 fragments (such as the GenBank for IAV:KM244070.1, its nucleotide sequence such as SEQ ID NO.142 It is shown) primer sets of amplification, N9 fragments (such as the GenBank for IAV:KC853231.1, its nucleotide sequence such as SEQ ID Shown in NO.143) primer sets of amplification, N10 fragments (such as the GenBank for IAV:CY103894.1, its nucleotide sequence As shown in SEQ ID NO.144) primer sets of amplification, N11 fragments (such as the GenBank for IAV:CY125947.1, its core Nucleotide sequence is as shown in SEQ ID NO.145) amplification primer sets,.

Wherein, the primer sets of the H1 areas fragment amplification for IAV include two pairs of primers, are respectively：H1-F3：Its core Nucleotide sequence is as shown in SEQ ID NO.1, H1-B3：Its nucleotide sequence is as shown in SEQ ID NO.2；And H1-FIP：Its Nucleotide sequence is as shown in SEQ ID NO.3, H1-BIP：Its nucleotide sequence is as shown in SEQ ID NO.4；

The primer sets of the H2 areas fragment amplification for IAV include two pairs of primers, are respectively：H2-F3：Its nucleotides sequence Row are as shown in SEQ ID NO.5, H2-B3：Its nucleotide sequence is as shown in SEQ ID NO.6；And H2-FIP：Its nucleotides Sequence is as shown in SEQ ID NO.7, H2-BIP：Its nucleotide sequence is as shown in SEQ ID NO.8；

The primer sets of the H3 areas fragment amplification for IAV include two pairs of primers, are respectively：H3-F3：Its nucleotides sequence Row are as shown in SEQ ID NO.9, H3-B3：Its nucleotide sequence is as shown in SEQ ID NO.10；And H3-FIP：Its nucleotides Sequence is as shown in SEQ ID NO.11, H3-BIP：Its nucleotide sequence is as shown in SEQ ID NO.12；

The primer sets of the H4 areas fragment amplification for IAV include two pairs of primers, are respectively：H4-F3：Its nucleotides sequence Row are as shown in SEQ ID NO.13, H4-B3：Its nucleotide sequence is as shown in SEQ ID NO.14；And H4-FIP：Its nucleosides Acid sequence is as shown in SEQ ID NO.15, H4-BIP：Its nucleotide sequence is as shown in SEQ ID NO.16；

The primer sets of the H5 areas fragment amplification for IAV include two pairs of primers, are respectively：H5-F3：Its nucleotides sequence Row are as shown in SEQ ID NO.17, H5-B3：Its nucleotide sequence is as shown in SEQ ID NO.18；And H5-FIP：Its nucleosides Acid sequence is as shown in SEQ ID NO.19, H5-BIP：Its nucleotide sequence is as shown in SEQ ID NO.20；

The primer sets of the H6 areas fragment amplification for IAV include two pairs of primers, are respectively：H6-F3：Its nucleotides sequence Row are as shown in SEQ ID NO.21, H6-B3：Its nucleotide sequence is as shown in SEQ ID NO.22；And H6-FIP：Its nucleosides Acid sequence is as shown in SEQ ID NO.23, H6-BIP：Its nucleotide sequence is as shown in SEQ ID NO.24；

The primer sets of the H7 areas fragment amplification for IAV include two pairs of primers, are respectively：H7-F3：Its nucleotides sequence Row are as shown in SEQ ID NO.25, H7-B3：Its nucleotide sequence is as shown in SEQ ID NO.26；And H7-FIP：Its nucleosides Acid sequence is as shown in SEQ ID NO.27, H7-BIP：Its nucleotide sequence is as shown in SEQ ID NO.28；

The primer sets of the H8 areas fragment amplification for IAV include two pairs of primers, are respectively：H8-F3：Its nucleotides sequence Row are as shown in SEQ ID NO.29, H8-B3：Its nucleotide sequence is as shown in SEQ ID NO.30；And H8-FIP：Its nucleosides Acid sequence is as shown in SEQ ID NO.31, H8-BIP：Its nucleotide sequence is as shown in SEQ ID NO.32；

The primer sets of the H9 areas fragment amplification for IAV include two pairs of primers, are respectively：H9-F3：Its nucleotides sequence Row are as shown in SEQ ID NO.33, H9-B3：Its nucleotide sequence is as shown in SEQ ID NO.34；And H9-FIP：Its nucleosides Acid sequence is as shown in SEQ ID NO.35, H9-BIP：Its nucleotide sequence is as shown in SEQ ID NO.36；

The primer sets of the H10 areas fragment amplification for IAV include two pairs of primers, are respectively：H10-F3：Its nucleotides Sequence is as shown in SEQ ID NO.37, H10-B3：Its nucleotide sequence is as shown in SEQ ID NO.38；And H10-FIP：Its Nucleotide sequence is as shown in SEQ ID NO.39, H10-BIP：Its nucleotide sequence is as shown in SEQ ID NO.40；

The primer sets of the H11 areas fragment amplification for IAV include two pairs of primers, are respectively：H11-F3：Its nucleotides Sequence is as shown in SEQ ID NO.41, H11-B3：Its nucleotide sequence is as shown in SEQ ID NO.42；And H11-FIP：Its Nucleotide sequence is as shown in SEQ ID NO.43, H11-BIP：Its nucleotide sequence is as shown in SEQ ID NO.44；

The primer sets of the H12 areas fragment amplification for IAV include two pairs of primers, are respectively：H12-F3：Its nucleotides Sequence is as shown in SEQ ID NO.45, H12-B3：Its nucleotide sequence is as shown in SEQ ID NO.46；And H12-FIP：Its Nucleotide sequence is as shown in SEQ ID NO.47, H12-BIP：Its nucleotide sequence is as shown in SEQ ID NO.48；

The primer sets of the H13 areas fragment amplification for IAV include two pairs of primers, are respectively：H13-F3：Its nucleotides Sequence is as shown in SEQ ID NO.49, H13-B3：Its nucleotide sequence is as shown in SEQ ID NO.50；And H13-FIP：Its Nucleotide sequence is as shown in SEQ ID NO.51, H13-BIP：Its nucleotide sequence is as shown in SEQ ID NO.52；

The primer sets of the H14 areas fragment amplification for IAV include two pairs of primers, are respectively：H14-F3：Its nucleotides Sequence is as shown in SEQ ID NO.53, H14-B3：Its nucleotide sequence is as shown in SEQ ID NO.54；And H14-FIP：Its Nucleotide sequence is as shown in SEQ ID NO.55, H14-BIP：Its nucleotide sequence is as shown in SEQ ID NO.56；

The primer sets of the H15 areas fragment amplification for IAV include two pairs of primers, are respectively：H15-F3：Its nucleotides Sequence is as shown in SEQ ID NO.57, H15-B3：Its nucleotide sequence is as shown in SEQ ID NO.58；And H15-FIP：Its Nucleotide sequence is as shown in SEQ ID NO.59, H15-BIP：Its nucleotide sequence is as shown in SEQ ID NO.60；

The primer sets of the H16 areas fragment amplification for IAV include two pairs of primers, are respectively：H16-F3：Its nucleotides Sequence is as shown in SEQ ID NO.61, H16-B3：Its nucleotide sequence is as shown in SEQ ID NO.62；And H16-FIP：Its Nucleotide sequence is as shown in SEQ ID NO.63, H16-BIP：Its nucleotide sequence is as shown in SEQ ID NO.64；

The primer sets of the H17 areas fragment amplification for IAV include two pairs of primers, are respectively：H17-F3：Its nucleotides Sequence is as shown in SEQ ID NO.65, H17-B3：Its nucleotide sequence is as shown in SEQ ID NO.66；And H17-FIP：Its Nucleotide sequence is as shown in SEQ ID NO.67, H17-BIP：Its nucleotide sequence is as shown in SEQ ID NO.68；

The primer sets of the H18 areas fragment amplification for IAV include two pairs of primers, are respectively：H18-F3：Its nucleotides Sequence is as shown in SEQ ID NO.69, H18-B3：Its nucleotide sequence is as shown in SEQ ID NO.70；And H18-FIP：Its Nucleotide sequence is as shown in SEQ ID NO.71, H18-BIP：Its nucleotide sequence is as shown in SEQ ID NO.72.

The nucleotide sequence of the above-mentioned each primer of the present invention is as shown in table 1.

The primer sequence of table 1

It is further preferred that LAMP primer group of the present invention includes the primer of H1~H18 areas fragment amplification for IAV One group in group or several groups, and, for the one or more in the primer sets of IAV N1~N11 areas fragment amplification.

The kit for being used to detect influenza A virus of the present invention, including above-mentioned LAMP primer group.It is preferred that by LAMP Working solution is made in primer sets, and the working solution mother liquor can be (20mM Tris-HCl pH8.8,10mM KCl, 10mM (NH₄)₂SO₄, 2~20mM MgSO₄, 0.1~0.5%Triton X-100,0.2~1M glycine betaines, 1~1.6mM dNTP) primer sets work It is FIP, BIP mixed liquor containing 1~20 μM F3, B3 and 5~80 μM to make liquid, and 10~15 μ L work is recommended when being detected The primer sets working solution for making liquid mother liquor and 3~5 μ L forms 25 μ L detection mixed liquor plus enzyme and water.

According to kit of the present invention, wherein, in addition to LAMP reaction solutions and LAMP nitrite ions.Further, institute Stating LAMP reaction solutions includes：Bst polymerases, AMV reverse transcriptase, LAMP reaction buffers and ultra-pure water.

Specifically, preferably, in kit of the present invention, including primer sets working solution (1~20 μM of F3, B3 and 5~80 μM of FIP, BIP mixed liquor), LAMP reaction solutions (20mM Tris-HCl pH8.8,10mM KCl, 10mM (NH₄)₂SO₄, 2~20mM MgSO₄, 0.1~0.5%Triton X-100,0.2~1M glycine betaines, 1~1.6mM dNTP, 8U Bst Archaeal dna polymerase and 5U AMV reverse transcriptases) and LAMP nitrite ions (hydroxynaphthol blue or eriochrome black T etc.).

LAMP reaction solutions (25 μ L systems)

20mM Tris-HCl pH8.8

10mM KCl

10mM(NH₄)₂SO₄

2~20mM MgSO₄

0.1~0.5%Triton X-100

0.2~1M glycine betaines

1~1.6mM dNTP

5~10U Bst archaeal dna polymerases (NEW ENGLAND Biolabs)

5~10U AMV reverse transcriptase (NEW ENGLAND Biolabs)

In preferable Sybr green I, the Eva green of LAMP nitrite ions, hydroxynaphthol blue (HNB), eriochrome black T (EBT) etc. One kind.

When being detected, such as 4~8 μ L primer sets working solution can be used to form 25 μ L inspection with LAMP reaction solutions Survey mixed liquor.LAMP nitrite ions addition can be 100~150 μm of ol/L detection mixed liquors.

Present invention also offers the IAV method for detecting virus of non-disease diagnostic purpose, comprise the following steps：

1) sample of nucleic acid, the working solution of the LAMP primer group and LAMP reaction solutions, ultra-pure water, LAMP sealing fluids is taken to mix Close, amplification reaction solution is made；

2) obtained amplification reaction solution, 60~65 DEG C of reactions 20~80min, preferably 63 DEG C reaction 60min, according to colour developing are taken As a result whether judgement sample contains IAV viruses.

According to right detection method of the present invention, wherein, in the working solution of the LAMP primer group, primer H1-F3 Concentration with H1-B3 is 0.2~0.4 μM, and primer H1-FIP and H1-BIP concentration are 1~2 μM；

The concentration of the primer H2-F3 and H2-B3 is 0.2~0.4 μM, and primer H2-FIP and H2-BIP concentration are 1~2 μM；

The concentration of the primer H3-F3 and H3-B3 is 0.2~0.4 μM, and primer H3-FIP and H3-BIP concentration are 1~2 μM；

The concentration of the primer H4-F3 and H4-B3 is 0.2~0.4 μM, and primer H4-FIP and H4-BIP concentration are 1~2 μM；

The concentration of the primer H5-F3 and H5-B3 is 0.2~0.4 μM, and primer H5-FIP and H5-BIP concentration are 1~2 μM；

The concentration of the primer H6-F3 and H6-B3 is 0.2~0.4 μM, and primer H6-FIP and H6-BIP concentration are 1~2 μM；

The concentration of the primer H73-F3 and H7-B3 is 0.2~0.4 μM, and primer H7FIP and H7-BIP concentration are 1~2 μM；

The concentration of the primer H8-F3 and H8-B3 is 0.2~0.4 μM, and primer H8-FIP and H8-BIP concentration are 1~2 μM；

The concentration of the primer H9-F3 and H9-B3 is 0.2~0.4 μM, and primer H9-FIP and H9-BIP concentration are 1~2 μM；

The concentration of the primer H10-F3 and H10-B3 is 0.2~0.4 μM, primer H10-FIP and H10-BIP concentration It is 1~2 μM；

The concentration of the primer H11-F3 and H11-B3 is 0.2~0.4 μM, primer H11-FIP and H11-BIP concentration It is 1~2 μM；

The concentration of the primer H12-F3 and H12-B3 is 0.2~0.4 μM, primer H12-FIP and H12-BIP concentration It is 1~2 μM；

The concentration of the primer H13-F3 and H13-B3 is 0.2~0.4 μM, primer H13-FIP and H13-BIP concentration It is 1~2 μM；

The concentration of the primer H14-F3 and H14-B3 is 0.2~0.4 μM, primer H14-FIP and H14-BIP concentration It is 1~2 μM；

The concentration of the primer H15-F3 and H15-B3 is 0.2~0.4 μM, primer H15-FIP and H15-BIP concentration It is 1~2 μM；

The concentration of the primer H16-F3 and H16-B3 is 0.2~0.4 μM, primer H16-FIP and H16-BIP concentration It is 1~2 μM；

The concentration of the primer H17-F3 and H17-B3 is 0.2~0.4 μM, primer H17-FIP and H17-BIP concentration It is 1~2 μM；

The concentration of the primer H18-F3 and H18-B3 is 0.2~0.4 μM, primer H18-FIP and H18-BIP concentration It is 1~2 μM；

The concentration of the primer N1-F3 and N1-B3 is 0.2~0.4 μM, and primer N1-FIP and N1-BIP concentration are 1~2 μM；

The concentration of the primer N2-F3 and N2-B3 is 0.2~0.4 μM, and primer N2-FIP and N2-BIP concentration are 1~2 μM；

The concentration of the primer N3-F3 and N3-B3 is 0.2~0.4 μM, and primer N3-FIP and N3-BIP concentration are 1~2 μM；

The concentration of the primer N4-F3 and N4-B3 is 0.2~0.4 μM, and primer N4-FIP and N4-BIP concentration are 1~2 μM；

The concentration of the primer N5-F3 and N5-B3 is 0.2~0.4 μM, and primer N5-FIP and N5-BIP concentration are 1~2 μM；

The concentration of the primer N6-F3 and N6-B3 is 0.2~0.4 μM, and primer N6-FIP and N6-BIP concentration are 1~2 μM；

The concentration of the primer N7-F3 and N7-B3 is 0.2~0.4 μM, and primer N7FIP and N7-BIP concentration are 1 ~2 μM；

The concentration of the primer N8-F3 and N8-B3 is 0.2~0.4 μM, and primer N8-FIP and N8-BIP concentration are 1~2 μM；

The concentration of the primer N9-F3 and N9-B3 is 0.2~0.4 μM, and primer N9-FIP and N9-BIP concentration are 1~2 μM；

The concentration of the primer N10-F3 and N10-B3 is 0.2~0.4 μM, primer N10-FIP and N10-BIP concentration It is 1~2 μM；

The concentration of the primer N11-F3 and N11-B3 is 0.2~0.4 μM, primer N11-FIP and N11-BIP concentration It is 1~2 μM；

Present invention also offers above-mentioned LAMP primer group and kit in the IAV Viral diagnosis of non-disease diagnostic purpose Application.

The present invention is based on LAMP technology, and 4 primers are separately designed for IAV 29 special conservative regions.The present invention carries The primer sets sensitiveness of confession is high, high specificity, and the kit being made from it, which can be detected fast and accurately in testing sample, to be contained Some IAV, and parting detection and identification can be carried out to the IAV types in testing sample.Due to primer sets provided by the invention It is short the time required to LAMP is expanded with high specificity, the time of detection is further shorten, simplifies operation.This hair It is bright by the use of artificial RNA as during control test, non-false positive and false negative phenomenon produce.Primer spirit provided by the invention Sensitivity and specificity are all very high, and the visualizing agent box provided will provide great convenience for Site Detection.Because the present invention carries The method or kit of confession can complete the quick and precisely detection to IAV without expensive instrument, without the operation of complexity, because This, when being broken out suitable for IAV, the field quick detection on the ground such as the not high airport of professional degree, customs, port, community.The present invention The visualizing agent box provided will provide great convenience for Site Detection, be prepared into can realizing for kit fast to IAV Fast accurately detection.

Brief description of the drawings

Fig. 1 is the curve map that the fluorescence intensity in the embodiment of the present invention 1 changes with the reaction time.

Fig. 2 is the curve map that the fluorescence intensity in the embodiment of the present invention 2 changes with the reaction time.

Fig. 3 is the curve map that the fluorescence intensity in the embodiment of the present invention 3 changes with the reaction time.

Fig. 4 is the curve map that the fluorescence intensity in the embodiment of the present invention 4 changes with the reaction time.

Fig. 5 is the curve map that the fluorescence intensity in the embodiment of the present invention 5 changes with the reaction time.

Fig. 6 is the curve map that the fluorescence intensity in the embodiment of the present invention 6 changes with the reaction time.

Fig. 7 is the curve map that the fluorescence intensity in the embodiment of the present invention 7 changes with the reaction time.

Fig. 8 is the curve map that the fluorescence intensity in the embodiment of the present invention 8 changes with the reaction time.

Fig. 9 is the curve map that the fluorescence intensity in the embodiment of the present invention 9 changes with the reaction time.

Figure 10 is the curve map that the fluorescence intensity in the embodiment of the present invention 10 changes with the reaction time.

Figure 11 is the curve map that the fluorescence intensity in the embodiment of the present invention 11 changes with the reaction time.

Figure 12 is the curve map that the fluorescence intensity in the embodiment of the present invention 12 changes with the reaction time.

Figure 13 is the curve map that the fluorescence intensity in the embodiment of the present invention 13 changes with the reaction time.

Figure 14 is the curve map that the fluorescence intensity in the embodiment of the present invention 14 changes with the reaction time.

Figure 15 is the curve map that the fluorescence intensity in the embodiment of the present invention 15 changes with the reaction time.

Figure 16 is the curve map that the fluorescence intensity in the embodiment of the present invention 16 changes with the reaction time.

Figure 17 is the curve map that the fluorescence intensity in the embodiment of the present invention 17 changes with the reaction time.

Figure 18 is the curve map that the fluorescence intensity in the embodiment of the present invention 18 changes with the reaction time.

Figure 19 is the curve map that the fluorescence intensity in the embodiment of the present invention 19 changes with the reaction time.

Figure 20 is the curve map that the fluorescence intensity in the embodiment of the present invention 20 changes with the reaction time.

Figure 21 is the curve map that the fluorescence intensity in the embodiment of the present invention 21 changes with the reaction time.

Figure 22 is the curve map that the fluorescence intensity in the embodiment of the present invention 22 changes with the reaction time.

Figure 23 is the curve map that the fluorescence intensity in the embodiment of the present invention 23 changes with the reaction time.

Figure 24 is the curve map that the fluorescence intensity in the embodiment of the present invention 24 changes with the reaction time.

Figure 25 is the curve map that the fluorescence intensity in the embodiment of the present invention 25 changes with the reaction time.

Figure 26 is the curve map that the fluorescence intensity in the embodiment of the present invention 26 changes with the reaction time.

Figure 27 is the curve map that the fluorescence intensity in the embodiment of the present invention 27 changes with the reaction time.

Figure 28 is the curve map that the fluorescence intensity in the embodiment of the present invention 28 changes with the reaction time.

Figure 29 is the curve map that the fluorescence intensity in the embodiment of the present invention 29 changes with the reaction time.

Figure 30 is the testing result schematic diagram in the embodiment of the present invention 30.

Embodiment

Experimental method used in following embodiments unless otherwise specified, is conventional method, specifically can refer to《Molecule Cloning experimentation guide》Listed specific method is carried out in the book of (third edition) J. Pehanorm Brookers one, or according to kit and production Product specification is carried out；Material used, reagent etc., unless otherwise specified, are commercially obtained in following embodiments.

Amplified reaction of the application Eva Green checkings of embodiment 1 to IAV-H1

Eva Green are similar with SYBR Green I, are a kind of to be incorporated into having for all dsDNA minor grooves regions The dyestuff of green excitation wavelength, its suppression to nucleic acid amplification reactions such as PCR are much smaller than the latter.Under free state, Eva Green sends faint fluorescence, but once combined with double-stranded DNA, fluorescence greatly enhances.Therefore, Eva Green fluorescence letter Number intensity is related to the quantity of double-stranded DNA, and double-stranded DNA number existing for nucleic acid amplification system can be detected according to fluorescence signal Amount.

Reaction solution combines (25 μ L)

20mM Tris-HCl pH8.8

10mM KCl

10mM(NH₄)₂SO₄

14mM MgSO₄

0.1%Triton X-100

1M glycine betaines

1.25mM dNTP

8U Bst archaeal dna polymerases (NEW ENGLAND Biolabs)

1X Eva Green(Biotum)

Primer：

200nM H1-F3/SEQ ID NO.1

200nM H1-B3/SEQ ID NO.2

1600nM H1-FIP/SEQ ID NO.3

1600nM H1-BIP/SEQ ID NO.4

Target:IAV-H1dsDNA/SEQ ID NO.117

Control group without target is set simultaneously.

It is 63 DEG C to set ABI StepOne real time PCR reaction temperatures constant, reaction time 60min.Fluorescence The curve that intensity changes with the reaction time is as shown in Figure 1.Fluoroscopic examination is applied to can wherein to realize the purpose monitored in real time, led to Judged result can be shifted to an earlier date by crossing real-time amplification curve.

Amplified reaction of the application Eva Green checkings of embodiment 2 to IAV-H2

Reaction solution combines (25 μ L), with embodiment 1；

Primer：

200nM H2-F3/SEQ ID NO.5

200nM H2-B3/SEQ ID NO.6

1600nM H2-FIP/SEQ ID NO.7

1600nM H2-BIP/SEQ ID NO.8

Target:IAV-H2dsDNA/SEQ ID NO.118

Control group without target is set simultaneously.

Amplified reaction of the application Eva Green checkings of embodiment 3 to IAV-H3

Reaction solution combines (25 μ L), with embodiment 1；

Primer：

200nM H3-F3/SEQ ID NO.9

200nM H3-B3/SEQ ID NO.10

1600nM H3-FIP/SEQ ID NO.11

1600nM H3-BIP/SEQ ID NO.12

Target:IAV-H3dsDNA/SEQ ID NO.119

Control group without target is set simultaneously.

Amplified reaction of the application Eva Green checkings of embodiment 4 to IAV-H4

Reaction solution combines (25 μ L), with embodiment 1；

Primer：

200nM H4-F3/SEQ ID NO.13

200nM H4-B3/SEQ ID NO.14

1600nM H4-FIP/SEQ ID NO.15

1600nM H4-BIP/SEQ ID NO.16

Target:IAV-H4dsDNA/SEQ ID NO.120

Control group without target is set simultaneously.

Amplified reaction of the application Eva Green checkings of embodiment 5 to IAV-H5

Reaction solution combines (25 μ L), with embodiment 1；

Primer：

200nM H5-F3/SEQ ID NO.17

200nM H5-B3/SEQ ID NO.18

1600nM H5-FIP/SEQ ID NO.19

1600nM H5-BIP/SEQ ID NO.20

Target:IAV-H5dsDNA/SEQ ID NO.121

Control group without target is set simultaneously.

Amplified reaction of the application Eva Green checkings of embodiment 6 to IAV-H6

Reaction solution combines (25 μ L), with embodiment 1；

Primer：

200nM H6-F3/SEQ ID NO.21

200nM H6-B3/SEQ ID NO.22

1600nM H6-FIP/SEQ ID NO.23

1600nM H6-BIP/SEQ ID NO.24

Target:IAV-H6dsDNA/SEQ ID NO.122

Control group without target is set simultaneously.

Amplified reaction of the application Eva Green checkings of embodiment 7 to IAV-H7

Reaction solution combines (25 μ L), with embodiment 1；

Primer：

200nM H7-F3/SEQ ID NO.25

200nM H7-B3/SEQ ID NO.26

1600nM H7-FIP/SEQ ID NO.27

1600nM H7-BIP/SEQ ID NO.28

Target:IAV-H7dsDNA/SEQ ID NO.123

Control group without target is set simultaneously.

Amplified reaction of the application Eva Green checkings of embodiment 8 to IAV-H8

Reaction solution combines (25 μ L), with embodiment 1；

Primer：

200nM H8-F3/SEQ ID NO.29

200nM H8-B3/SEQ ID NO.30

1600nM H8-FIP/SEQ ID NO.31

1600nM H8-BIP/SEQ ID NO.32

Target:IAV-H8dsDNA/SEQ ID NO.124

Control group without target is set simultaneously.

Amplified reaction of the application Eva Green checkings of embodiment 9 to IAV-H9

Reaction solution combines (25 μ L), with embodiment 1；

Primer：

200nM H9-F3/SEQ ID NO.33

200nM H9-B3/SEQ ID NO.34

1600nM H9-FIP/SEQ ID NO.35

1600nM H9-BIP/SEQ ID NO.36

Target:IAV-H9dsDNA/SEQ ID NO.125

Control group without target is set simultaneously.

Amplified reaction of the application Eva Green checkings of embodiment 10 to IAV-H10

Reaction solution combines (25 μ L), with embodiment 1；

Primer：

200nM H10-F3/SEQ ID NO.37

200nM H10-B3/SEQ ID NO.38

1600nM H10-FIP/SEQ ID NO.39

1600nM H10-BIP/SEQ ID NO.40

Target:IAV-H10dsDNA/SEQ ID NO.126

Control group without target is set simultaneously.

Amplified reaction of the application Eva Green checkings of embodiment 11 to IAV-H11

Reaction solution combines (25 μ L), with embodiment 1；

Primer：

200nM H11-F3/SEQ ID NO.41

200nM H11-B3/SEQ ID NO.42

1600nM H11-FIP/SEQ ID NO.43

1600nM H11-BIP/SEQ ID NO.44

Target:IAV-H11dsDNA/SEQ ID NO.127

Control group without target is set simultaneously.

Amplified reaction of the application Eva Green checkings of embodiment 12 to IAV-H12

Reaction solution combines (25 μ L), with embodiment 1；

Primer：

200nM H12-F3/SEQ ID NO.45

200nM H12-B3/SEQ ID NO.46

1600nM H12-FIP/SEQ ID NO.47

1600nM H12-BIP/SEQ ID NO.48

Target:IAV-H12dsDNA/SEQ ID NO.128

Control group without target is set simultaneously.

Amplified reaction of the application Eva Green checkings of embodiment 13 to IAV-H13

Reaction solution combines (25 μ L), with embodiment 1；

Primer：

200nM H13-F3/SEQ ID NO.49

200nM H13-B3/SEQ ID NO.50

1600nM H13-FIP/SEQ ID NO.51

1600nM H13-BIP/SEQ ID NO.52

Target:IAV-H13dsDNA/SEQ ID NO.129

Control group without target is set simultaneously.

Amplified reaction of the application Eva Green checkings of embodiment 14 to IAV-H14

Reaction solution combines (25 μ L), with embodiment 1；

Primer：

200nM H14-F3/SEQ ID NO.53

200nM H14-B3/SEQ ID NO.54

1600nM H14-FIP/SEQ ID NO.55

1600nM H14-BIP/SEQ ID NO.56

Target:IAV-H14dsDNA/SEQ ID NO.130

Control group without target is set simultaneously.

Amplified reaction of the application Eva Green checkings of embodiment 15 to IAV-H15

Reaction solution combines (25 μ L), with embodiment 1；

Primer：

200nM H15-F3/SEQ ID NO.57

200nM H15-B3/SEQ ID NO.58

1600nM H15-FIP/SEQ ID NO.59

1600nM H15-BIP/SEQ ID NO.60

Target:IAV-H15dsDNA/SEQ ID NO.131

Control group without target is set simultaneously.

Amplified reaction of the application Eva Green checkings of embodiment 16 to IAV-H16

Reaction solution combines (25 μ L), with embodiment 1；

Primer：

200nM H16-F3/SEQ ID NO.61

200nM H16-B3/SEQ ID NO.62

1600nM H16-FIP/SEQ ID NO.63

1600nM H16-BIP/SEQ ID NO.64

Target:IAV-H16dsDNA/SEQ ID NO.132

Control group without target is set simultaneously.

Amplified reaction of the application Eva Green checkings of embodiment 17 to IAV-H17

Reaction solution combines (25 μ L), with embodiment 1；

Primer：

200nM H17-F3/SEQ ID NO.65

200nM H17-B3/SEQ ID NO.66

1600nM H17-FIP/SEQ ID NO.67

1600nM H17-BIP/SEQ ID NO.68

Target:IAV-H17dsDNA/SEQ ID NO.133

Control group without target is set simultaneously.

Amplified reaction of the application Eva Green checkings of embodiment 18 to IAV-H18

Reaction solution combines (25 μ L), with embodiment 1；

Primer：

200nM H18-F3/SEQ ID NO.69

200nM H18-B3/SEQ ID NO.70

1600nM H18-FIP/SEQ ID NO.71

1600nM H18-BIP/SEQ ID NO.72

Target:IAV-H18dsDNA/SEQ ID NO.134

Control group without target is set simultaneously.

Amplified reaction of the application Eva Green checkings of embodiment 19 to IAV-N1

Reaction solution combines (25 μ L), with embodiment 1；

Primer：

200nM N1-F3/SEQ ID NO.73

200nM N1-B3/SEQ ID NO.74

1600nM N1-FIP/SEQ ID NO.75

1600nM N1-BIP/SEQ ID NO.76

Target:IAV-N1dsDNA/SEQ ID NO.135

Control group without target is set simultaneously.

Amplified reaction of the application Eva Green checkings of embodiment 20 to IAV-N2

Reaction solution combines (25 μ L), with embodiment 1；

Primer：

200nM N2-F3/SEQ ID NO.77

200nM N2-B3/SEQ ID NO.78

1600nM N2-FIP/SEQ ID NO.79

1600nM N2-BIP/SEQ ID NO.80

Target:IAV-N2dsDNA/SEQ ID NO.136

Control group without target is set simultaneously.

Amplified reaction of the application Eva Green checkings of embodiment 21 to IAV-N3

Reaction solution combines (25 μ L), with embodiment 1；

Primer：

200nM N3-F3/SEQ ID NO.81

200nM N3-B3/SEQ ID NO.82

1600nM N3-FIP/SEQ ID NO.8 3

1600nM N3-BIP/SEQ ID NO.84

Target:IAV-N3dsDNA/SEQ ID NO.137

Control group without target is set simultaneously.

Amplified reaction of the application Eva Green checkings of embodiment 22 to IAV-N4

Reaction solution combines (25 μ L), with embodiment 1；

Primer：

200nM N4-F3/SEQ ID NO.85

200nM N4-B3/SEQ ID NO.86

1600nM N4-FIP/SEQ ID NO.87

1600nM N4-BIP/SEQ ID NO.88

Target:IAV-N4dsDNA/SEQ ID NO.138

Control group without target is set simultaneously.

Amplified reaction of the application Eva Green checkings of embodiment 23 to IAV-N5

Reaction solution combines (25 μ L), with embodiment 1；

Primer：

200nM N5-F3/SEQ ID NO.89

200nM N5-B3/SEQ ID NO.90

1600nM N5-FIP/SEQ ID NO.91

1600nM N5-BIP/SEQ ID NO.92

Target:IAV-N5dsDNA/SEQ ID NO.139

Control group without target is set simultaneously.

Amplified reaction of the application Eva Green checkings of embodiment 24 to IAV-N6

Reaction solution combines (25 μ L), with embodiment 1；

Primer：

200nM N6-F3/SEQ ID NO.93

200nM N6-B3/SEQ ID NO.94

1600nM N6-FIP/SEQ ID NO.95

1600nM N6-BIP/SEQ ID NO.96

Target:IAV-N6dsDNA/SEQ ID NO.140

Control group without target is set simultaneously.

Amplified reaction of the application Eva Green checkings of embodiment 25 to IAV-N7

Reaction solution combines (25 μ L), with embodiment 1；

Primer：

200nM N7-F3/SEQ ID NO.97

200nM N7-B3/SEQ ID NO.98

1600nM N7-FIP/SEQ ID NO.99

1600nM N7-BIP/SEQ ID NO.100

Target:IAV-N7dsDNA/SEQ ID NO.141

Control group without target is set simultaneously.

Amplified reaction of the application Eva Green checkings of embodiment 26 to IAV-N8

Reaction solution combines (25 μ L), with embodiment 1；

Primer：

200nM N8-F3/SEQ ID NO.101

200nM N8-B3/SEQ ID NO.102

1600nM N8-FIP/SEQ ID NO.103

1600nM N8-BIP/SEQ ID NO.104

Target:IAV-N8dsDNA/SEQ ID NO.142

Control group without target is set simultaneously.

Amplified reaction of the application Eva Green checkings of embodiment 27 to IAV-N9

Reaction solution combines (25 μ L), with embodiment 1；

Primer：

200nM N9-F3/SEQ ID NO.105

200nM N9-B3/SEQ ID NO.106

1600nM N9-FIP/SEQ ID NO.107

1600nM N9-BIP/SEQ ID NO.108

Target:IAV-N9dsDNA/SEQ ID NO.143

Control group without target is set simultaneously.

Amplified reaction of the application Eva Green checkings of embodiment 28 to IAV-N10

Reaction solution combines (25 μ L), with embodiment 1；

Primer：

200nM N10-F3/SEQ ID NO.109

200nM N10-B3/SEQ ID NO.110

1600nM N10-FIP/SEQ ID NO.111

1600nM N10-BIP/SEQ ID NO.112

Target:IAV-N10dsDNA/SEQ ID NO.144

Control group without target is set simultaneously.

Amplified reaction of the application Eva Green checkings of embodiment 29 to IAV-N11

Reaction solution combines (25 μ L), with embodiment 1；

Primer：

200nM N11-F3/SEQ ID NO.113

200nM N11-B3/SEQ ID NO.114

1600nM N11-FIP/SEQ ID NO.115

1600nM N11-BIP/SEQ ID NO.116

Target:IAV-N11dsDNA/SEQ ID NO.145

Control group without target is set simultaneously.

The Visual retrieval of the artificial RNA fragments of embodiment 30IAV gene target fragments

Addition reverse transcriptase can realize the detection for RNA in LAMP reaction systems.Because IAV is strong RNA diseases Poison, there is extremely strong harmfulness, must be carried out for the viral research in high-level biosafety laboratory, therefore for the disease The research and development of malicious detection technique and product are typically simulated using harmless artificial RNA fragments.Therefore this patent also uses artificial RNA Fragment, the RiboMaxTMLarge Scale RNA Production Systems provided using Promega companies carry out external Transcribe to obtain.Then RNA detection researchs are carried out using institute's visualizing agent.

Hydroxynaphthol blue (HNB) is the indicator for being applied to Visual retrieval in LAMP technology.Its principle is：When generation core A large amount of insoluble accessory substance magnesium pyrophosphates can be produced during sour amplification in vitro reaction, so as to cause magnesium ion to reduce, and HNB is magnesium ion Indicator thus can monitor whether there is amplified reaction.When amplified reaction does not occur for LAMP, magnesium ion concentration is high in system, reaction Liquid is in purple；When amplified reaction occurs for LAMP, magnesium ion concentration is low in system, and reaction solution is in blueness.

The combination for the reaction solution for carrying out IAV viral RNA detections by these primers and visualizing agent is shown below.

Reaction solution combines (25 μ L)

20mM Tris-HCl pH8.8

10mM KCl

10mM(NH₄)₂SO₄

14mM MgSO₄

0.1%Triton X-100

1M glycine betaines

1.25mM dNTP

8U Bst archaeal dna polymerases (NEW ENGLAND Biolabs)

5U AMV reverse transcriptase (NEW ENGLAND Biolabs)

120μM HNB

Above by the shared reagent of Visual retrieval, following is for primer and target sequence used in each RNA fragments Row：

1) IAV-H1RNA detection primers：

200nM H1-F3/SEQ ID NO.1

200nM H1-B3/SEQ ID NO.2

1600nM H1-FIP/SEQ ID NO.3

1600nM H1-BIP/SEQ ID NO.4

Target:IAV-H1dsDNA/SEQ ID NO.117

Control group without target is set simultaneously.

2) IAV-H2RNA detection primers：

200nM H2-F3/SEQ ID NO.5

200nM H2-B3/SEQ ID NO.6

1600nM H2-FIP/SEQ ID NO.7

1600nM H2-BIP/SEQ ID NO.8

Target:IAV-H2dsDNA/SEQ ID NO.118

Control group without target is set simultaneously.

3) IAV-H3RNA detection primers：

200nM H3-F3/SEQ ID NO.9

200nM H3-B3/SEQ ID NO.10

1600nM H3-FIP/SEQ ID NO.11

1600nM H3-BIP/SEQ ID NO.12

Target:IAV-H3dsDNA/SEQ ID NO.119

Control group without target is set simultaneously.

4) IAV-H4RNA detection primers：

200nM H4-F3/SEQ ID NO.13

200nM H4-B3/SEQ ID NO.14

1600nM H4-FIP/SEQ ID NO.15

1600nM H4-BIP/SEQ ID NO.16

Target:IAV-H4dsDNA/SEQ ID NO.120

Control group without target is set simultaneously.

5) IAV-H5RNA detection primers：

200nM H5-F3/SEQ ID NO.17

200nM H5-B3/SEQ ID NO.18

1600nM H5-FIP/SEQ ID NO.18

1600nM H5-BIP/SEQ ID NO.20

Target:IAV-H5dsDNA/SEQ ID NO.121

Control group without target is set simultaneously.

6) IAV-H6RNA detection primers：

200nM H6-F3/SEQ ID NO.21

200nM H6-B3/SEQ ID NO.22

1600nM H6-FIP/SEQ ID NO.23

1600nM H6-BIP/SEQ ID NO.2 4

Target:IAV-H6dsDNA/SEQ ID NO.122

Control group without target is set simultaneously.

7) IAV-H7RNA detection primers：

200nM H7-F3/SEQ ID NO.25

200nM H7-B3/SEQ ID NO.26

1600nM H7-FIP/SEQ ID NO.27

1600nM H7-BIP/SEQ ID NO.28

Target:IAV-H7dsDNA/SEQ ID NO.123

Control group without target is set simultaneously.

8) IAV-H8RNA detection primers：

200nM H8-F3/SEQ ID NO.29

200nM H8-B3/SEQ ID NO.30

1600nM H8-FIP/SEQ ID NO.31

1600nM H8-BIP/SEQ ID NO.32

Target:IAV-H8dsDNA/SEQ ID NO.124

Control group without target is set simultaneously.

9) IAV-H9RNA detection primers：

200nM H9-F3/SEQ ID NO.33

200nM H9-B3/SEQ ID NO.34

1600nM H9-FIP/SEQ ID NO.35

1600nM H9-BIP/SEQ ID NO.36

Target:IAV-H9dsDNA/SEQ ID NO.125

Control group without target is set simultaneously.

10) IAV-H10RNA detection primers：

200nM H10-F3/SEQ ID NO.37

200nM H10-B3/SEQ ID NO.38

1600nM H10-FIP/SEQ ID NO.39

1600nM H10-BIP/SEQ ID NO.40

Target:IAV-H10dsDNA/SEQ ID NO.126

Control group without target is set simultaneously.

11) IAV-H11RNA detection primers：

200nM H11-F3/SEQ ID NO.41

200nM H11-B3/SEQ ID NO.42

1600nM H11-FIP/SEQ ID NO.43

1600nM H11-BIP/SEQ ID NO.44

Target:IAV-H11dsDNA/SEQ ID NO.127

Control group without target is set simultaneously.

12) IAV-H12RNA detection primers：

200nM H12-F3/SEQ ID NO.45

200nM H12-B3/SEQ ID NO.46

1600nM H12-FIP/SEQ ID NO.47

1600nM H12-BIP/SEQ ID NO.48

Target:IAV-H12dsDNA/SEQ ID NO.128

Control group without target is set simultaneously.

13) IAV-H13RNA detection primers：

200nM H13-F3/SEQ ID NO.49

200nM H13-B3/SEQ ID NO.50

1600nM H13-FIP/SEQ ID NO.51

1600nM H13-BIP/SEQ ID NO.52

Target:IAV-H13dsDNA/SEQ ID NO.129

Control group without target is set simultaneously.

14) IAV-H14RNA detection primers：

200nM H14-F3/SEQ ID NO.53

200nM H14-B3/SEQ ID NO.54

1600nM H14-FIP/SEQ ID NO.55

1600nM H14-BIP/SEQ ID NO.56

Target:IAV-H14dsDNA/SEQ ID NO.130

Control group without target is set simultaneously.

15) IAV-H15RNA detection primers：

200nM H15-F3/SEQ ID NO.57

200nM H15-B3/SEQ ID NO.58

1600nM H15-FIP/SEQ ID NO.59

1600nM H15-BIP/SEQ ID NO.60

Target:IAV-H15dsDNA/SEQ ID NO.131

Control group without target is set simultaneously.

16) IAV-H16RNA detection primers：

200nM H16-F3/SEQ ID NO.61

200nM H16-B3/SEQ ID NO.62

1600nM H16-FIP/SEQ ID NO.63

1600nM H16-BIP/SEQ ID NO.64

Target:IAV-H16dsDNA/SEQ ID NO.132

Control group without target is set simultaneously.

17) IAV-H17RNA detection primers：

200nM H17-F3/SEQ ID NO.65

200nM H17-B3/SEQ ID NO.66

1600nM H17-FIP/SEQ ID NO.67

1600nM H17-BIP/SEQ ID NO.68

Target:IAV-H17dsDNA/SEQ ID NO.133

Control group without target is set simultaneously.

18) IAV-H18RNA detection primers：

200nM H18-F3/SEQ ID NO.69

200nM H18-B3/SEQ ID NO.70

1600nM H18-FIP/SEQ ID NO.71

1600nM H18-BIP/SEQ ID NO.72

Target:IAV-H18dsDNA/SEQ ID NO.134

Control group without target is set simultaneously.

19) IAV-N1RNA detection primers：

200nM N1-F3/SEQ ID NO.73

200nM N1-B3/SEQ ID NO.74

1600nM N1-FIP/SEQ ID NO.75

1600nM N1-BIP/SEQ ID NO.76

Target:IAV-N1dsDNA/SEQ ID NO.135

Control group without target is set simultaneously.

20) IAV-N2RNA detection primers：

200nM N2-F3/SEQ ID NO.77

200nM N2-B3/SEQ ID NO.78

1600nM N2-FIP/SEQ ID NO.79

1600nM N2-BIP/SEQ ID NO.80

Target:IAV-N2dsDNA/SEQ ID NO.136

Control group without target is set simultaneously.

21) IAV-N3RNA detection primers：

200nM N3-F3/SEQ ID NO.81

200nM N3-B3/SEQ ID NO.82

1600nM N3-FIP/SEQ ID NO.8 3

1600nM N3-BIP/SEQ ID NO.84

Target:IAV-N3dsDNA/SEQ ID NO.137

Control group without target is set simultaneously.

22) IAV-N4RNA detection primers：

200nM N4-F3/SEQ ID NO.85

200nM N4-B3/SEQ ID NO.86

1600nM N4-FIP/SEQ ID NO.87

1600nM N4-BIP/SEQ ID NO.88

Target:IAV-N4dsDNA/SEQ ID NO.138

Control group without target is set simultaneously.

23) IAV-N5RNA detection primers：

200nM N5-F3/SEQ ID NO.89

200nM N5-B3/SEQ ID NO.90

1600nM N5-FIP/SEQ ID NO.91

1600nM N5-BIP/SEQ ID NO.92

Target:IAV-N5dsDNA/SEQ ID NO.139

Control group without target is set simultaneously.

24) IAV-N6RNA detection primers：

200nM N6-F3/SEQ ID NO.93

200nM N6-B3/SEQ ID NO.94

1600nM N6-FIP/SEQ ID NO.95

1600nM N6-BIP/SEQ ID NO.96 targets:IAV-N6dsDNA/SEQ ID NO.140 are set without target simultaneously Control group.

25) IAV-N7RNA detection primers：

200nM N7-F3/SEQ ID NO.97

200nM N7-B3/SEQ ID NO.98

1600nM N7-FIP/SEQ ID NO.99

1600nM N7-BIP/SEQ ID NO.100 targets:IAV-N7dsDNA/SEQ ID NO.141 are set without target simultaneously Control group.

26) IAV-N8RNA detection primers：

200nM N8-F3/SEQ ID NO.101

200nM N8-B3/SEQ ID NO.102

1600nM N8-FIP/SEQ ID NO.103

1600nM N8-BIP/SEQ ID NO.104 targets:IAV-N8dsDNA/SEQ ID NO.142 are set without target simultaneously Control group.

27) IAV-N9RNA detection primers：

200nM N9-F3/SEQ ID NO.105

200nM N9-B3/SEQ ID NO.106

1600nM N9-FIP/SEQ ID NO.107

1600nM N9-BIP/SEQ ID NO.108

Target:IAV-N9dsDNA/SEQ ID NO.143

Control group without target is set simultaneously.

28) IAV-N10RNA detection primers：

200nM N10-F3/SEQ ID NO.109

200nM N10-B3/SEQ ID NO.110

1600nM N10-FIP/SEQ ID NO.111

D600nM N10-BIP/SEQ ID NO.112 targets:IAV-N10dsDNA/SEQ ID NO.144 are set without target simultaneously Control group.

29) IAV-N11RNA detection primers：

200nM N11-F3/SEQ ID NO.113

200nM N11-B3/SEQ ID NO.114

1600nM N11-FIP/SEQ ID NO.115

1600nM N11-BIP/SEQ ID NO.116

Target:IAV-N11dsDNA/SEQ ID NO.145

Control group without target is set simultaneously.

As a result judge：Blueness is the positive, and purple is feminine gender.As a result show in the figure 7, the reference numerals difference of each reaction tube Counter sample is as follows：

1：The positive findings of IAV H1RNA detections.2：The negative findings of IAV H1RNA detections.3：IAV H2RNA detections Positive findings.4：The negative findings of IAV H2RNA detections.5：The positive findings of IAV H3RNA detections.6：IAV H3RNA are detected Negative findings.7：The positive findings of IAV H4RNA detections.8：The negative findings of IAV H4RNA detections.9：IAV H5RNA are examined The positive findings of survey.10：The negative findings of IAV H5RNA detections.11：The positive findings of IAV H6RNA detections.12：IAV The negative findings of H6RNA detections.13：The positive findings of IAV H7RNA detections.14：The negative findings of IAV H7RNA detections.15： The positive findings of IAV H8RNA detections.16：The negative findings of IAV H8RNA detections.17：The positive knot of IAV H9RNA detections Fruit.18：The negative findings of IAV H9RNA detections.19：The positive findings of IAV H10RNA detections.20：IAV H10RNA detections Negative findings.21：The positive findings of IAV H11RNA detections.22：The negative findings of IAV H11RNA detections.23：IAV The positive findings of H12RNA detections.24：The negative findings of IAV H12RNA detections.25：The positive findings of IAV H13RNA detections. 26：The negative findings of IAV H13RNA detections.27：The positive findings of IAV H14RNA detections.28：The moon of IAV H14RNA detections Property result.29：The positive findings of IAV H15RNA detections.30：The negative findings of IAV H15RNA detections.31：IAV H16RNA The positive findings of detection.32：The negative findings of IAV H16RNA detections.33：The positive findings of IAV H17RNA detections.34：IAV The negative findings of H17RNA detections.35：The positive findings of IAV H18RNA detections.36：The negative findings of IAV H18RNA detections. 37：The positive findings of IAV N1RNA detections.38：The negative findings of IAV N1RNA detections.39：The positive of IAV N2RNA detections As a result.40：The negative findings of IAV N1RNA detections.41：The positive findings of IAV N3RNA detections.42：IAV N3RNA detections Negative findings.43：The positive findings of IAV N4RNA detections.44：The negative findings of IAV N4RNA detections.45：IAV N5RNA are examined The positive findings of survey.46：The negative findings of IAV N5RNA detections.47：The positive findings of IAV N6RNA detections.48：IAV The negative findings of N6RNA detections.49：The positive findings of IAV N7RNA detections.50：The negative findings of IAV N7RNA detections.51： The positive findings of IAV N8RNA detections.52：The negative findings of IAV N8RNA detections.53：The positive knot of IAV N9RNA detections Fruit.54：The negative findings of IAV N9RNA detections.55：The positive findings of IAV N10RNA detections.56：IAV N10RNA detections Negative findings.57：The positive findings of IAV N11RNA detections.58：The negative findings of IAV N11RNA detections.

Certainly, the present invention can also have various embodiments, in the case of without departing substantially from spirit of the invention and its essence, be familiar with Those skilled in the art can be made according to disclosure of the invention it is various it is corresponding change and deformation, but these it is corresponding change and Deformation should all belong to the scope of the claims of the present invention.

Sequence table

<110>Chinese Academy Of Sciences Process Engineering Research Institute

<120>A kind of LAMP primer group and kit for being used to expand influenza A virus

<160> 145

<170> SIPOSequenceListing 1.0

<210> 1

<211> 24

<212> DNA

<213> Artificial

<400> 1

actattattg gacactatta gacc 24

<210> 2

<211> 21

<212> DNA

<213> Artificial

<400> 2

gtacattctg aaaaggaagg c 21

<210> 3

<211> 43

<212> DNA

<213> Artificial

<400> 3

tgcgaaagca taccatggtg ggagacacta taacatttga agc 43

<210> 4

<211> 43

<212> DNA

<213> Artificial

<400> 4

tggaattata acatcagatg ctccagttca aagccccatg agg 43

<210> 5

<211> 18

<212> DNA

<213> Artificial

<400> 5

aatacgattg ggcgagaa 18

<210> 6

<211> 19

<212> DNA

<213> Artificial

<400> 6

tcctattggg aaccgaatc 19

<210> 7

<211> 46

<212> DNA

<213> Artificial

<400> 7

tgcaaaggac cagcaattat tattgaagac gttatagtca cgagag 46

<210> 8

<211> 41

<212> DNA

<213> Artificial

<400> 8

cttgcccaag gggcattact gatctatagg gtgttctgtc t 41

<210> 9

<211> 19

<212> DNA

<213> Artificial

<400> 9

gcaaaagagg acctgctaa 19

<210> 10

<211> 18

<212> DNA

<213> Artificial

<400> 10

actctccctg atgcttga 18

<210> 11

<211> 41

<212> DNA

<213> Artificial

<400> 11

cacgttcagc actgggtatg ttcttcagta gactgaactg g 41

<210> 12

<211> 47

<212> DNA

<213> Artificial

<400> 12

gacaaactat acatctgggg agtacacata caggttggtt tgttctt 47

<210> 13

<211> 17

<212> DNA

<213> Artificial

<400> 13

acccctgcgc aaaatgc 17

<210> 14

<211> 19

<212> DNA

<213> Artificial

<400> 14

ttgttagggt tccgcgtcc 19

<210> 15

<211> 42

<212> DNA

<213> Artificial

<400> 15

agaagtacca cctgggagcc aggtattggc ggagacagga ca 42

<210> 16

<211> 39

<212> DNA

<213> Artificial

<400> 16

gaactggacc cgaagcagca gtggcgccat cttcatgga 39

<210> 17

<211> 18

<212> DNA

<213> Artificial

<400> 17

ggcaaagtgg aagaatgg 18

<210> 18

<211> 21

<212> DNA

<213> Artificial

<400> 18

ggcatactag agtttatcgc c 21

<210> 19

<211> 49

<212> DNA

<213> Artificial

<400> 19

ttgtatgcat attctggagc aatgaagttc ttctggacaa ttttaaagc 49

<210> 20

<211> 41

<212> DNA

<213> Artificial

<400> 20

gtcaagaaag gggactcagc acattggagt ttgacacttg g 41

<210> 21

<211> 20

<212> DNA

<213> Artificial

<400> 21

gtgatgctgt ctgccaaacg 20

<210> 22

<211> 20

<212> DNA

<213> Artificial

<400> 22

tcatccctgt ccatcctcct 20

<210> 23

<211> 42

<212> DNA

<213> Artificial

<400> 23

tggggcattc tccaatccag aggttgctgg agcactaagg ac 42

<210> 24

<211> 41

<212> DNA

<213> Artificial

<400> 24

gcctaagact ggcaactggt ctcagctatg gccccgaaca a 41

<210> 25

<211> 19

<212> DNA

<213> Artificial

<400> 25

accagagatt ccatgacag 19

<210> 26

<211> 19

<212> DNA

<213> Artificial

<400> 26

agtcgtcatc acacttgtg 19

<210> 27

<211> 42

<212> DNA

<213> Artificial

<400> 27

ccagatcaat tgtgtgctgg ttcaagtgtg gtcctataac gc 42

<210> 28

<211> 43

<212> DNA

<213> Artificial

<400> 28

atgaacaaac tatacgaacg agtgagaagc aaccagtgcc atc 43

<210> 29

<211> 19

<212> DNA

<213> Artificial

<400> 29

gtgttaccct ggatctgtg 19

<210> 30

<211> 20

<212> DNA

<213> Artificial

<400> 30

tcataagtgt ctggttcctt 20

<210> 31

<211> 49

<212> DNA

<213> Artificial

<400> 31

tccacctgga atagtcaaat agtctaaatc tagaagaact gaggtttgt 49

<210> 32

<211> 43

<212> DNA

<213> Artificial

<400> 32

gactagatct ggaacgagta aagcggtcaa ccaattgatg ctc 43

<210> 33

<211> 20

<212> DNA

<213> Artificial

<400> 33

caatcaacaa actccaccga 20

<210> 34

<211> 20

<212> DNA

<213> Artificial

<400> 34

cgacctttca acgatgtagg 20

<210> 35

<211> 42

<212> DNA

<213> Artificial

<400> 35

ttgcacacaa catcccattg tgcagagagc aatgttcctg ta 42

<210> 36

<211> 43

<212> DNA

<213> Artificial

<400> 36

atcccctcat tctagacacc tgcctcctag tagtatgtca cag 43

<210> 37

<211> 18

<212> DNA

<213> Artificial

<400> 37

tgagacggta gagagcac 18

<210> 38

<211> 21

<212> DNA

<213> Artificial

<400> 38

attattttct gcctcaatgc t 21

<210> 39

<211> 43

<212> DNA

<213> Artificial

<400> 39

aacattccta ccgggtgaca attgtgtatg aaaggaagaa gct 43

<210> 40

<211> 39

<212> DNA

<213> Artificial

<400> 40

cacctgtttg tgatccgcac tggataacag tgggcaatg 39

<210> 41

<211> 21

<212> DNA

<213> Artificial

<400> 41

ctggatcatt ctgttcaatc a 21

<210> 42

<211> 22

<212> DNA

<213> Artificial

<400> 42

tcttagttct tcttcactct ct 22

<210> 43

<211> 45

<212> DNA

<213> Artificial

<400> 43

cacacatagg gtttcctaat atccatggaa aacaaccaat aagcc 45

<210> 44

<211> 44

<212> DNA

<213> Artificial

<400> 44

actaattgga aagacttcat ggtctttcct gggtaacaga ttcc 44

<210> 45

<211> 21

<212> DNA

<213> Artificial

<400> 45

atggttgacc ttaaaatcag g 21

<210> 46

<211> 18

<212> DNA

<213> Artificial

<400> 46

gttgtcctct cacgagtg 18

<210> 47

<211> 45

<212> DNA

<213> Artificial

<400> 47

atggcccagg tgaatacaat gtcacaattt ccagtccaaa cagat 45

<210> 48

<211> 44

<212> DNA

<213> Artificial

<400> 48

ctgatactct ctcttcagtc accaggccct atattaggct tgaa 44

<210> 49

<211> 18

<212> DNA

<213> Artificial

<400> 49

agtcaccagc tccattga 18

<210> 50

<211> 18

<212> DNA

<213> Artificial

<400> 50

gtagcaaagc ccatgagg 18

<210> 51

<211> 39

<212> DNA

<213> Artificial

<400> 51

tgcactggac tgactccatt tctgatcgag acaaaccac 39

<210> 52

<211> 42

<212> DNA

<213> Artificial

<400> 52

agattgcagc tttgaaggat ggatcaggta tgaccactct ct 42

<210> 53

<211> 18

<212> DNA

<213> Artificial

<400> 53

gccccttgaa gcttgtcg 18

<210> 54

<211> 20

<212> DNA

<213> Artificial

<400> 54

gcttgctagg atgcttctga 20

<210> 55

<211> 40

<212> DNA

<213> Artificial

<400> 55

gtcctgcaaa cggtcacagc cgcaagactg ccacctcatc 40

<210> 56

<211> 42

<212> DNA

<213> Artificial

<400> 56

tcattgaaag gcccactgca gtactctggt aatctgggac gt 42

<210> 57

<211> 18

<212> DNA

<213> Artificial

<400> 57

agctgggaac aagctgat 18

<210> 58

<211> 19

<212> DNA

<213> Artificial

<400> 58

gggctatgaa tgcaccatt 19

<210> 59

<211> 42

<212> DNA

<213> Artificial

<400> 59

ccattcactt tgggcctgtc cagtaggaag ctcaaaatac ca 42

<210> 60

<211> 42

<212> DNA

<213> Artificial

<400> 60

ccgggaggat cgactttcat gaaggtaaaa gtgactgtat cc 42

<210> 61

<211> 19

<212> DNA

<213> Artificial

<400> 61

tggggaatcc accatccag 19

<210> 62

<211> 19

<212> DNA

<213> Artificial

<400> 62

gaaggcctcc attgctctc 19

<210> 63

<211> 42

<212> DNA

<213> Artificial

<400> 63

gcgtttgctc cactcttttg tcgattcgga agcaacagcc ac 42

<210> 64

<211> 41

<212> DNA

<213> Artificial

<400> 64

cgggaccaga ataggtgacg gaaacgttat cctttcgccg g 41

<210> 65

<211> 22

<212> DNA

<213> Artificial

<400> 65

gacagaatat gcataggcta tc 22

<210> 66

<211> 18

<212> DNA

<213> Artificial

<400> 66

aataaccacc cggctaga 18

<210> 67

<211> 43

<212> DNA

<213> Artificial

<400> 67

gctccggtaa ctggaacatt ctgcaaacca aaataaccaa acg 43

<210> 68

<211> 46

<212> DNA

<213> Artificial

<400> 68

aatactagaa accaatcaca acggatgcat gattgtaggt ccaatg 46

<210> 69

<211> 25

<212> DNA

<213> Artificial

<400> 69

gagaaaataa tagacatgac cagat 25

<210> 70

<211> 20

<212> DNA

<213> Artificial

<400> 70

tgttctgcag cattagatgt 20

<210> 71

<211> 42

<212> DNA

<213> Artificial

<400> 71

aaggaagcac catttgtgtc actgatgtga ctacaaacaa cg 42

<210> 72

<211> 46

<212> DNA

<213> Artificial

<400> 72

taaactgggt gcagcaaaac aaagcttatc aaaagtgggt tgcaaa 46

<210> 73

<211> 23

<212> DNA

<213> Artificial

<400> 73

aattaccact atgaagagtg ttc 23

<210> 74

<211> 18

<212> DNA

<213> Artificial

<400> 74

taccccatat gccccatt 18

<210> 75

<211> 45

<212> DNA

<213> Artificial

<400> 75

cgatttgaac catgccaatt gtcctgttat cctgatgcta gtgag 45

<210> 76

<211> 39

<212> DNA

<213> Artificial

<400> 76

cagcggagtt tttggagaca aatactggac cacaactgc 39

<210> 77

<211> 23

<212> DNA

<213> Artificial

<400> 77

aatatggcag attatagcat tga 23

<210> 78

<211> 20

<212> DNA

<213> Artificial

<400> 78

tccttgttga ttgttcttcc 20

<210> 79

<211> 41

<212> DNA

<213> Artificial

<400> 79

ttgctgctgc tagagctatc gcaattatgt gtgctcagga c 41

<210> 80

<211> 41

<212> DNA

<213> Artificial

<400> 80

tgcaaggatc ccaataacga gacatccaaa cgtcatttcc a 41

<210> 81

<211> 22

<212> DNA

<213> Artificial

<400> 81

gacagactcc attaaatcat gg 22

<210> 82

<211> 23

<212> DNA

<213> Artificial

<400> 82

caagtgttgt atctttgttt tgg 23

<210> 83

<211> 45

<212> DNA

<213> Artificial

<400> 83

gtaacagcga caacacaggt ccatactaag aacccaagaa tctga 45

<210> 84

<211> 43

<212> DNA

<213> Artificial

<400> 84

gatggccctg cggctaatag ttattacctt tccttctcgt atc 43

<210> 85

<211> 19

<212> DNA

<213> Artificial

<400> 85

gtgacccaaa cacatgaac 19

<210> 86

<211> 22

<212> DNA

<213> Artificial

<400> 86

ccattgtctt tacttagagg tg 22

<210> 87

<211> 42

<212> DNA

<213> Artificial

<400> 87

gcacattggt gacattgaca aagaaccctg ttcgaacaac ac 42

<210> 88

<211> 45

<212> DNA

<213> Artificial

<400> 88

taccacagta attgaaccct cagcttattg ggcacaagtc tcttc 45

<210> 89

<211> 19

<212> DNA

<213> Artificial

<400> 89

tgccattagg atcttcacc 19

<210> 90

<211> 18

<212> DNA

<213> Artificial

<400> 90

acctcaccac atctgttg 18

<210> 91

<211> 40

<212> DNA

<213> Artificial

<400> 91

ttgccatcat ggcatgctgt caatgcttat caggccaagt 40

<210> 92

<211> 39

<212> DNA

<213> Artificial

<400> 92

gtggctggca gtagggataa catcccccca taatggatt 39

<210> 93

<211> 20

<212> DNA

<213> Artificial

<400> 93

agggtcgagt gcataggatg 20

<210> 94

<211> 20

<212> DNA

<213> Artificial

<400> 94

gggccatctg tcatgactac 20

<210> 95

<211> 40

<212> DNA

<213> Artificial

<400> 95

gatgcattgt tgttcgctcc cggcacgtca tgccacgatg 40

<210> 96

<211> 42

<212> DNA

<213> Artificial

<400> 96

agaaattcca tcatgggccg gaccccttat ggcacacaca tt 42

<210> 97

<211> 20

<212> DNA

<213> Artificial

<400> 97

gcactgcatc aaggaacaac 20

<210> 98

<211> 20

<212> DNA

<213> Artificial

<400> 98

attgtggtag tgagccttcg 20

<210> 99

<211> 41

<212> DNA

<213> Artificial

<400> 99

ttgctgacta caggggggct cacggaacaa tacacgacag g 41

<210> 100

<211> 40

<212> DNA

<213> Artificial

<400> 100

aagcaccagt tgccatgacg gactgtagct gttgcgttgt 40

<210> 101

<211> 19

<212> DNA

<213> Artificial

<400> 101

agtgcaggac gttcttcct 19

<210> 102

<211> 19

<212> DNA

<213> Artificial

<400> 102

agggcccgtt actccaatt 19

<210> 103

<211> 41

<212> DNA

<213> Artificial

<400> 103

taggggctcc ggtctttcac tacccagggt tccttactca a 41

<210> 104

<211> 42

<212> DNA

<213> Artificial

<400> 104

tcacccaatg tgtaccaggc aaccatttct tcccatcgtg gc 42

<210> 105

<211> 18

<212> DNA

<213> Artificial

<400> 105

gactggaact gctaagca 18

<210> 106

<211> 20

<212> DNA

<213> Artificial

<400> 106

aagggtcatt acacttacct 20

<210> 107

<211> 39

<212> DNA

<213> Artificial

<400> 107

ttgagccctg ccaattgtcc gaatgctcat gttacggga 39

<210> 108

<211> 42

<212> DNA

<213> Artificial

<400> 108

agtagcaatg acacacacta gtcaatttgg gtcattcggt cg 42

<210> 109

<211> 19

<212> DNA

<213> Artificial

<400> 109

aggaaaaagg agaaggagg 19

<210> 110

<211> 20

<212> DNA

<213> Artificial

<400> 110

gggttgaaaa aatccgcttc 20

<210> 111

<211> 45

<212> DNA

<213> Artificial

<400> 111

acccattctg agataactca gtcttccaag gtttcatact tgacg 45

<210> 112

<211> 46

<212> DNA

<213> Artificial

<400> 112

tagaacaaat ccctgacggg acggttctct tattagaaaa aagttc 46

<210> 113

<211> 22

<212> DNA

<213> Artificial

<400> 113

tgtaggaaat aggaactacc aa 22

<210> 114

<211> 18

<212> DNA

<213> Artificial

<400> 114

catctggcat tgagcctc 18

<210> 115

<211> 47

<212> DNA

<213> Artificial

<400> 115

cttgacaatc acgtgattca taagcttaag taacagcaca attggga 47

<210> 116

<211> 46

<212> DNA

<213> Artificial

<400> 116

gtgtttttgg attgaaattg ctgcagtaat cagatcattg gatgac 46

<210> 117

<211> 981

<212> DNA

<213> Artificial

<400> 117

gacaccatct gtgtgggcta ccatgcaaac aactctacag acactgttga cacagtactg 60

gaaaagaatg tgaccgtgac tcactcagtg aatttgctcg aagacagcca taatgggaaa 120

ctctgcagcc taaacgggat acctccccta caactgggaa agtgcaatgt ggcgggatgg 180

ctcctgggca atccagagtg tgatcttcta ctcactgcaa actcatggtc ctacataata 240

gaaacttcaa actcagaaaa cggaacatgc taccccggtg aattcataga ttatgaagaa 300

ttaagagagc agctaagttc agtttcttca tttgaaaaat ttgaaatttt cccgaaggca 360

agctcatggc caaatcatga gacaactaaa ggtgttacag ctgcatgctc ttactctgga 420

gccagcagtt tttaccggaa tttgctgtgg ataacaaaga aagggacttc atatccaaaa 480

ctcagcaaat catacacgaa caataagggg aaagaagtgc ttgtgctctg gggggtgcac 540

caccctccaa gtgtcagtga gcaacaaagt ctataccaga atgctgatgc atacgtttca 600

gttggatcgt caaaatacaa ccgaagattc gctccggaaa tagcagctag acctaaagtt 660

agaggacagg caggcagaat gaactattat tggacactat tagaccaagg agacactata 720

acatttgaag ccactgggaa tttgatagca ccatggtatg ctttcgcatt gaataagggg 780

tctgactctg gaattataac atcagatgct ccagttcaca attgtgacac aaggtgccaa 840

acccctcatg gggctttgaa cagcagcctt ccttttcaga atgtacatcc tatcactatt 900

ggagaatgtc ccaaatacgt caagagcacc aaactaagaa tggcaacagg actaagaaat 960

gtcccatcca ttcagtccag a 981

<210> 118

<211> 1410

<212> DNA

<213> Artificial

<400> 118

atgaatccaa atcagaagat aataacaatt ggtgtagtga acactactct atcaacaata 60

gcccttctta ttggagtggg aaacttgatt ttcaacactg tcatacatga gaaaataggg 120

gaccactcga ctgtagtgta cccaacgata acaaccccgg tggtaccaaa ctgcagtgac 180

accataataa catacaacaa cacagtaata aacaacataa caacaacaat aataaccgaa 240

gcagaaagac attttaagcc ctcactgccg ctgtgtccct tcagaggatt tttccctttt 300

cataaggata atgcaatacg attgggcgag aacaaagacg ttatagtcac gagagagcct 360

tacgttagtt gcgacaataa taattgctgg tcctttgcac ttgcccaagg ggcattactg 420

ggaacccaac acagcaatgg aaccattaaa gacagaacac cctatagatc attgattcgg 480

ttcccaatag gaacagcccc ggtactaggg aattacaaag agatatgcat agcttggtca 540

agtagcagct gtttcgacgg gaaggagtgg atgcatgtct gcatgacagg gaacgacaat 600

gatgcaagtg ctcagataat atatgcaggg aaaatgacag actccatcaa atcatggaga 660

agagacatac taagaaccca agagtctgaa tgtcaatgta ttgacgggac ctgtgttgtc 720

gctgttacag atggacctgc tgctaatagt gcagaccata gggtttattg gatacgagaa 780

ggaaaggtga taaagtatga aaacattccc aaaacgaaga tacagcactt ggaggagtgt 840

tcttgttatg tggacattga cgtatactgt gtatgtagag acaattggaa aggttccaac 900

aggccttgga tgagaatcaa caatgagacc atattggaga ccgggtatgt gtgtagcaaa 960

tttcattcgg acacccccag accagccgac ccttcaacag tatcatgtga ttccccaagt 1020

aatgtaaatg gaggacccgg agttaagggg tttggcttca aatccggcaa tgacgtatgg 1080

ttggggagga ctgtatcaac tagcggtaga tcaggctttg aaatcatcaa agtcacagaa 1140

gggtggatca attctcccag ccatgccaaa tcagtcacac aaacattagt gtcaaacaat 1200

gattggtcag gttattcagg tagtttcatt gttgaaaaca acggttgttt ccagccttgc 1260

ttctatattg aacttatacg agggaggccc aataagaatg atgacgtttc ctggacaagc 1320

aatagtatag ttactttctg tggactagac aatgaacctg gatcgggaaa ttggcctgat 1380

ggttccaaca ttgggttcat gcccaagtaa 1410

<210> 119

<211> 1765

<212> DNA

<213> Artificial

<400> 119

agcaaaagca ggggatactt tcattaatca tgaagaccgt tattgcttta agctacattc 60

tctgtctgac tttcggacag gacctcccag ggaatgacaa cagtacagca acactgtgcc 120

tggggcacca tgcagtgccg aatgggacaa tagtgaagac aatcacagat gatcagattg 180

aggtgactaa tgctactgag ctagttcaaa gctcctcaac agggaaaata tgcaacaatc 240

ctcacaggat ccttgatgga agggcctgca cattaataga tgctctactg ggggatcctc 300

attgcgatgt ctttcaaaat gagacgtggg acctttttgt ggagcgaagc aatgctttca 360

gcaactgtta cccttatgat ataccagatt atgcatccct taggtcccta gttgcctcat 420

caggcacatt ggagttcatc actgagggtt tcacctggac aggagtaact cagaatggag 480

ggagcagtgc ttgcaaaaga ggacctgcta acggtttctt cagtagactg aactggttga 540

ctaaatcaga aagcgcatac ccagtgctga acgtgactat gccaaataat gacaattttg 600

acaaactata catctgggga gtacaccacc cgagcacaaa tcaagaacaa accaacctgt 660

atgttcaagc atcagggaga gtcacagtct ctaccaggag aagtcagcag actataatcc 720

cgaatattgg atctagaccc tgggtaaggg gccagcctgg cagaataagc atctattgga 780

caatagttaa acctggggac gtgctggtaa tcaacagtaa tggaaaccta atcgctcctc 840

ggggttactt caagatgcgc actgggaaaa gctcaataat gaggtcagat gcacctattg 900

acacctgtat ctctgagtgc atcactccaa atggaagcat tcccaatgac aagcccttcc 960

aaaatgtaaa caagatcaca tacggagcat gtcccaagta tgttaagcag aacaccctga 1020

agttggcaac agggatgcgg aatgtaccag agaaacaaac cagaggccta ttcggtgcaa 1080

tagcaggttt tatagaaaat ggatgggagg gaatgataga tggctggtat ggcttcaggc 1140

atcaaaattc tgagggtaca ggacaagcag cagaccttaa aagcactcag gcagccattg 1200

accaaatcaa taggaaattg aacagagtga ttgaaaagac gaatgagaag ttccatcaaa 1260

tcgaaaagga attctccgaa gtagaaggga ggattcagga ccttgagaaa tacgttgaag 1320

acacgaaaat agatctctgg tcttacaatg cggaacttct tgttgcccta gagaatcagc 1380

atacaatcga tctggctgat tcagaaatga acaaattatt tgaaaaaacc aggaggcaac 1440

tgagggaaaa tgctgaagac atgggcaatg gttgtttcaa gatataccac aaatgtgaca 1500

atgcttgcat agagtcaatt agaaacggga cttatgatca tgatatatac agagacgagg 1560

cattgaacaa ccggttccag atcaaaggtg tcgaactaaa atctggatac aaagactgga 1620

tcctgtggat ttcctttgcc atatcatgcc ttttgctttg tgttgttttg ctgggtttca 1680

ttatgtgggc ctgccagaga ggcaacatta ggtgcaacat ttgcatttga gtatactaat 1740

gattaaaaac acccttgttt ctact 1765

<210> 120

<211> 1695

<212> DNA

<213> Artificial

<400> 120

atgctatcaa ttgtgatttt gtttctgctc atagcagaga actcttccca aaactacaca 60

ggaaaccctg tgatatgcat gggacatcat gctgtggcca atgggactat ggtaaagacc 120

ctagccgatg atcaagtgga agtggtcact gcacaggaac tggtagaatc acaaaatctc 180

ccggaactat gtccgagccc tctgagacta gttgacggcc agacctgtga tatcatcaat 240

ggagcattgg ggagcccagg gtgtgaccat ttgaatggtg cagaatggga cgttttcata 300

gagaggccca atgcagtaga cacttgctat ccatttgatg tgccagagta ccagagtctg 360

agaagcatac tcgccaacaa tgggaaattc gaattcattg ccgaagaatt ccaatggaac 420

acggtgaagc aaaatggaaa atccggagcc tgcaagaggg caaatgtgga tgatttcttt 480

aacaggctaa actggctggt gaagtcagat gggaatgcgt acccccttca gaatttgaca 540

aagataaaca atggggatta tgcaaggctt tacatctggg gagttcatca cccttcgaca 600

agcacagagc aaaccaacct gtacaagaat aaccctggaa gagtcactgt gtctaccaaa 660

actagtcaaa caagtgtagt gcctgacatt ggcagcagac ccttggtgag aggacaaagt 720

ggcagagtga gtttctactg gactattgta gagcctggag acttgatagt cttcaacaca 780

atagggaatc taattgcccc gagaggccat tacaagttaa acaatcagaa gaagagcaca 840

attctaaata ctgcaattcc cataggctca tgtgtcagca aatgtcacac agacaaaggt 900

tctctctcta caactaagcc cttccaaaat atctcgagga tagcagttgg agactgcccc 960

agatatgtca aacagggctc tctaaaactt gcaacaggga tgaggaacat tcctgaaaag 1020

gcatcaagag ggctttttgg agcaatagct gggttcatag agaatggctg gcaaggtcta 1080

atcgatggtt ggtatggatt cagacaccag aatgcagaag gaacaggaac agcggcagat 1140

ctcaaatcca ctcaggcagc cattgatcaa atcaatggga aacttaaccg tcttattgag 1200

aagacaaacg ataaatacca tcaaatcgaa aaagagttcg agcaagttga aggaagaatt 1260

caagatctgg aaaactatgt tgaggacaca aagattgatt tatggtcata taatgcagag 1320

ctattagtcg ctctagaaaa ccaacacact atagacgtga ctgattcaga aatgaacaaa 1380

ctctttgaaa gagtaaggcg tcaactcaga gagaatgctg aagacaaagg gaatgggtgt 1440

ttcgaaatat tccacaagtg tgataacaac tgcattgaaa gtattcggaa tgggacttat 1500

gatcatgata tttatagaga tgaagcaatc aacaatcgat tccaaatcca gggagtcaaa 1560

ttgacccagg gatataagga catcattctt tggatttcat tctccatatc atgctttttg 1620

ctcgtagcac tacttttagc cttcattttg tgggcttgtc agaacggaaa catccggtgc 1680

cagatttgta tttag 1695

<210> 121

<211> 1760

<212> DNA

<213> Artificial

<400> 121

gcaggggtat aatctgtcaa aatggagaaa atagtgcttc ttcttgcaat agtcagtctt 60

gtcaaaagtg atcagatttg cattggttac catgcaaaca actcgacaga gcaggttgac 120

acaataatgg aaaagaacgt tactgttaca catgcccaag acatactgga aaagacacac 180

aatgggaagc tctgcgatct aaatggagtg aagcctctca ttttgagaga ttgtagtgta 240

gctggatggc tcctcggaaa ccctatgtgt gacgaattca tcaatgtgcc ggaatggtct 300

tacatagtgg agaaggccag tccagccaat gacctctgtt acccagggga tttcaacgac 360

tatgaagaac tgaaacacct attgagcaga acaaaccatt ttgagaaaat tcagatcatc 420

cccaaaagtt cttggtccaa tcatgatgcc tcatcagggg tgagctcagc atgtccatac 480

catgggaggt cctccttttt cagaaatgtg gtatggctta tcaaaaagaa cagtgcatac 540

ccaacaataa agaggagcta caataatacc aaccaagaag atcttttagt actgtggggg 600

attcaccatc ctaatgatgc ggcagagcag acaaagctct atcaaaaccc aaccacttac 660

atttccgttg gaacatcaac actgaaccag agattggttc cagaaatagc tactagaccc 720

aaagtaaacg ggcaaagtgg aagaatggag ttcttctgga caattttaaa gccgaatgat 780

gccatcaatt tcgagagtaa tggaaatttc attgctccag aatatgcata caaaattgtc 840

aagaaagggg actcagcaat tatgaaaagt gaattggaat atggtaactg caacaccaag 900

tgtcaaactc caatgggggc gataaactct agtatgccat tccacaacat acaccccctc 960

accatcgggg aatgccccaa atatgtgaaa tcaaacagat tagtccttgc gactggactc 1020

agaaataccc ctcagagaga gagaagaaga aaaaagagag gactatttgg agctatagca 1080

ggttttatag agggaggatg gcagggaatg gtagatggtt ggtatgggta ccaccatagc 1140

aatgagcagg ggagtggata cgctgcagac aaagaatcca ctcaaaaggc aatagatgga 1200

gtcaccaata aggtcaactc gatcattgac aaaatgaaca ctcagtttga ggccgttgga 1260

agggaattta ataacttgga aaggaggata gagaatttaa acaagcagat ggaagacgga 1320

ttcctagatg tctggactta taatgctgaa cttctggttc tcatggaaaa tgagagaact 1380

ctagactttc atgactcaaa tgtcaagaac ctttatgaca aggtccgact acagcttagg 1440

gataatgcaa aggagctggg taatggttgt ttcgagttct atcacaaatg tgataatgaa 1500

tgtatggaaa gtgtaaaaaa cggaacgtat gactacccgc agtattcaga agaagcaaga 1560

ctaaacagag aggaaataag tggagtaaaa ttggaatcaa tgggaactta ccaaatactg 1620

tcaatttatt caacagtggc gagttcccta gcactggcaa tcatggtagc tggtctatct 1680

ttatggatgt gctccaatgg atcgttacaa tgcagaattt gcatttaaat ttgtgagttc 1740

agattgtagt taaaaacacc 1760

<210> 122

<211> 1745

<212> DNA

<213> Artificial

<400> 122

agcaaaagca ggggaaaatg attgcaatca taatacttgc gatagtggcc tctaccagca 60

aatcagacaa gatctgcatt ggataccatg ccaacaactc gacaacgcaa gtggacacaa 120

tattagagaa gaatgtgaca gtgacgcact cagttgagct cctagagagt caaaaggagg 180

aaagattctg cagagtgtta aataaagcac ctctggatct aaagggttgc accattgaag 240

ggtggattct tggaaacccc caatgtgaca tcttgcttgg ggaccaacgt tggtcttaca 300

tagtggagag acctggagcc caaaatggga tatgctaccc agggatattg aatgaattag 360

aagagctaaa agcactcatt gggtctggag aaagagtaca gaggtttgaa atgtttccca 420

agagcacgtg ggccggagta gacactagca ggggagttac gaaagcttgc ccctacatta 480

gtggatcgtc tttctacgga aatcttttgt ggataataaa aacggagtct gctgcatacc 540

cagtaattaa gggaacatac aataacactg gctcccagcc gatcctgtat ttctggggtg 600

tacaccatcc tccagatacc aatgagcaaa ataccttgta tggctctggt gacagatatg 660

ttaggatggg aactgagagc atgaattttg ccaaaagtcc ggaaatagca gcaagaccag 720

ctgtgaatgg gcaaaggggg cgaattgact attattggtc tgtgttgaag ccaggagaaa 780

ccttaaatgt agaatccaat ggcaatttaa tagctccttg gtatgcttac aaattcacca 840

gttccaacaa taagggagct gtcttcaaat caaaccttcc aattgaaaat tgtgatgctg 900

tctgccaaac ggttgctgga gcactaagga caaacaaaac tttccaaaat gttagccccc 960

tctggattgg agaatgcccc aagtatgtta aaagtgatag cctaagactg gcaactggtc 1020

tgagaaatgt cccacaggca gagacaagag gattgttcgg ggccatagct gggtttatag 1080

aaggaggatg gacagggatg attgacggct ggtacggata ccaccatgag aattcacagg 1140

gatcgggtta tgcagcagat aaagaaagta ctcagaaagc aattgacggg atcaccaata 1200

aagtcaattc catcattgac aagatgaaca cacagtttga ggcagtagac catgaattct 1260

caaatctcga aagaagagta gacaatctaa acaaaagaat ggaagatgga tttttggatg 1320

tgtggacata caatgctgaa cttttagttc tactggaaaa tgagagaacc ttagacctgc 1380

acgatgccaa tgtgaagaac ctatatgaaa aggtgaaatc acaattgaga gacaacgcga 1440

aggatttggg caatgggtgt tttgagtttt ggcataaatg tgatgatgaa tgcatcaact 1500

cagttaaaaa tggcacgtat gactacccaa aataccaaga cgagagcaaa cttaatagac 1560

aggaaataga ttcagtgaaa ctggaaaatc tgggtgtata tcaaattctt gctatttata 1620

gtacggtatc gagcggtcta gttttggtgg ggctgatcat tgccatgggt ctttggatgt 1680

gctcaaatgg ctcaatgcca tgcaagatat gtatataatt agaaaaaaac acccttgttt 1740

ctact 1745

<210> 123

<211> 805

<212> DNA

<213> Artificial

<400> 123

taataagtaa tttgcccttt cagaacataa atagcagggc agtagggaaa tgtccgagat 60

atgttaagca agagagtctg ctgctggcaa cagggatgaa gaatgttccc gaaattccaa 120

agggaagagg cctatttggt gctatagcgg gtttcattga aaatggatgg gaaggtctga 180

ttgatgggtg gtatggcttc aggcatcaaa atgcacaagg ggagggaact gctgcagatt 240

acaaaagcac ccaatcagca attgatcaag taacaggaaa attgaaccgg cttatagaaa 300

aaactaacca acaatttaag ttaatagaca atgaattcac tgaggttgaa aagcaaattg 360

gcaatgtgat aaattggacc agagattcca tgacagaagt gtggtcctat aacgctgaac 420

tcttggtagc aatggagaac cagcacacaa ttgatctggc cgactcagaa atgaacaaac 480

tatacgaacg agtgaagaga caactgagag agaatgctga agaagatggc actggttgct 540

tcgaaatatt tcacaagtgt gatgacgact gtatggccag tattagaaac aacacctatg 600

atcacagcaa gtacagggaa gaggcaatgc aaaatagaat acagattgac ccagtcaaac 660

taagcagcgg ctacaaagat gtgatacttt ggtttagctt cggggcatca tgtttcatac 720

ttctggccat tgcaatgggc cttgtcttca tatgtgtgaa gaatggaaac atgcggtgca 780

ctatttgtat ataagtttgg aaaaa 805

<210> 124

<211> 1698

<212> DNA

<213> Artificial

<400> 124

atggaaaaat tcatcgcaat agcaaccttg gcgagcacaa atgcatacga taggatatgc 60

attgggtacc aatcaaacaa ctccacagac acagtgaaca ctctcataga acagaatgta 120

ccagtcaccc aaacaatgga gctcgtggaa acagagaaac atcccgctta ttgtaacact 180

gatttaggtg ccccattgga actgcgagac tgcaagattg aggcagtaat ctatgggaac 240

cccaagtgtg acatccatct gaaggatcaa ggttggtcat acatagtgga gaggcccagc 300

gcaccagaag ggatgtgtta ccctggatct gtggaaaatc tagaagaact gaggtttgtc 360

ttctccagtg ctgcatctta caagagaata agactatttg actattccag gtggaatgtg 420

actagatctg gaacgagtaa agcatgcaat gcatcaacag gtggccaatc cttctatagg 480

agcatcaatt ggttgaccaa aaaggaacca gacacttatg acttcaatga aggagcttat 540

gttaataatg aagatggaga catcattttc ttatggggga tccatcatcc gccggacaca 600

aaagagcaga caacactata taaaaatgca aacactttga gtagtgttac tactaacact 660

ataaacagaa gctttcaacc aaatattggt cccagaccat tagtaagagg acagcaaggg 720

aggatggatt actattgggg cattctgaaa agaggggaga ctctgaagat caggaccaac 780

ggaaatttaa tcgcacctga atttggctat ctgctcaaag gtgaaagcta cggcagaata 840

attcaaaatg aggatatacc catcgggaac tgtaacacaa aatgtcaaac atatgcggga 900

gcaatcaata gcagcaaacc ctttcagaat gcaagtaggc attacatggg agaatgtccc 960

aaatatgtga agaaggcaag cttgcgactt gcagttgggc ttaggaatac gccttctgtt 1020

gaacccagag gactgtttgg agccattgct ggtttcattg aaggaggatg gtctggaatg 1080

attgatgggt ggtatggatt tcatcacagc aattcagagg gaacaggaat ggcagctgac 1140

cagaaatcaa cacaagaagc catcgataag atcaccaata aagtcaacaa tatagttgac 1200

aagatgaaca gggagtttga agttgtgaat catgagttct ctgaagttga aaaaagaata 1260

aacatgataa acgataaaat agatgaccaa attgaagatc tttgggctta caatgcagag 1320

ctccttgtgc tcttagagaa ccagaaaacg ctagacgaac atgattccaa tgtcaaaaac 1380

ctttttgatg aagtgaaaag gagactgtca gccaatgcaa tagatgctgg gaacggttgc 1440

tttgacatac ttcacaaatg cgacaatgag tgtatggaaa ctataaagaa cggaacttac 1500

gatcataagg aatatgaaga ggaggctaaa ctagaaagga gcaagataaa tggagtaaaa 1560

ctagaagaga acaccactta caaaattctt agcatttaca gtacagtggc ggccagtctt 1620

tgcttggcaa tcctgattgc tggaggttta atcctgggca tgcaaaatgg atcttgtaga 1680

tgcatgttct gtatttga 1698

<210> 125

<211> 1683

<212> DNA

<213> Artificial

<400> 125

atggaaacaa aagcaataat tgctgcactg ctaatggtaa cagcagccaa tgctgataaa 60

atctgcattg gctatcaatc aacaaactcc accgaaactg tcgacacact aacagagagc 120

aatgttcctg taacacacac taaagaattg ctccacacag aacacaatgg gatgttgtgt 180

gcaactgatc taggacatcc cctcattcta gacacctgca ctattgaagg actaatctat 240

ggaaatcctt cctgtgacat actactagga ggaaaagaat ggtcctacat cgttgaaagg 300

tcgtcagccg tcaatggaat gtgttaccca gggaatgtag agaacctaga agagcttagg 360

tcacttttca gttctgcaaa atcttacaaa agaatccaaa tctttccaga taaaacttgg 420

aatgtaacat acagcggaac aagcagagca tgctcaaatt cattctatag gagcatgaga 480

tggctgaccc acaagagtaa ttcctaccct tttcagaacg ctcattacac caacaatgag 540

agggagaaca ttcttttcat gtggggcata caccatccac ctaccgacac agaacagaca 600

gacttataca agaatgccga tacaacaaca agtgtgacaa cagaagatat aaatcgcact 660

ttcaaaccag taatagggcc aaggcccctt gtcaacggcc agcagggaag aatcgattac 720

tattggtctg tactaaaacc tgggcagact ttgagaataa ggtccaatgg aaatctaatt 780

gccccttggt atggacatgt tcttacggga gagagccatg gaagaatcct aaagactgat 840

ttgaacaatg gtaattgtgt ggtacaatgt cagactgaaa aaggaggact caatacaacc 900

ttgccatttc acaatattag caaatatgca tttgggaatt gtcctaagta tgttggagtg 960

aaaagtctca aattgccggt tggtttaagg aatgtgcctg ctgtatctag tagaggactg 1020

ttcggagcaa tagctgggtt catagaggga ggttggccag ggctagtggc cggatggtat 1080

ggttttcaac attcaaacga tcagggagtt gggatggccg cagacaaggg atctacccaa 1140

aaagcaatcg ataaaataac atcaaaagtg aacaacataa tcgataaaat gaacaagcaa 1200

tatgaggtca ttgatcatga attcaatgag cttgaggcca ggctgaacat gatcaacaac 1260

aaaattgatg atcagattca agacatttgg gcttataatg cagagttgct ggtcttgctt 1320

gaaaaccaga aaactctaga tgagcatgat gcaaatgtga acaacctata caacaaagtg 1380

aaaagagcct tgggttccaa tgctgtagag gatggaaacg gatgctttga actgtaccac 1440

aaatgtgatg atcaatgcat ggaaacaatc agaaatggaa cctacgacag gcagaaatac 1500

caagaagaat caagattaga aaggcagaaa atagaagggg taaagctgga atctgagggc 1560

acttacaaga tcctcaccat ttattcgact gtcgcctcat ctcttgtgct tgcaatgggg 1620

tttgctgcct tcctattctg ggccatgtcc aatggctctt gcagatgcaa catttgtata 1680

taa 1683

<210> 126

<211> 1727

<212> DNA

<213> Artificial

<400> 126

agcaaaagca ggggtcacaa tgtacaaagt agtagtaata attgcgctcc ttggagcagt 60

gaaaggtctt gacagaatct gcctaggaca ccatgcggtt gccaatggaa ccattgtgaa 120

gacccttaca aatgaacaag aggaagtgac caatgctact gagacggtag agagcacaaa 180

tttgaataaa ttgtgtatga aaggaagaag ctacaaggac ttgggcaatt gtcacccggt 240

aggaatgttg ataggaacac ctgtttgtga tccgcacttg accgggacct gggacactct 300

cattgagcga gagaatgcca ttgcccactg ttatccaggg gcaaccataa atgaagaagc 360

attgaggcag aaaataatgg aaagtggagg aatcagcaag atgagcactg gcttcactta 420

tgggtcttcc atcacctcag ctgggaccac taaggcatgc atgagaaatg gaggagatag 480

tttctatgca gagctcaaat ggctagtgtc aaagacaaag ggacaaaatt tccctcagac 540

aacaaacacc tatcggaata cggacacagc agaacatctc ataatatggg gaattcatca 600

cccttccagc acacaggaaa agaatgactt atacggaact cagtcactat ctatatcagt 660

tgagagttct acatatcaga acaactttgt tccagttgtt ggggcaagac ctcaggtcaa 720

tggacaaagt gggcgaattg actttcactg gacactagta cagccgggtg acaacataac 780

cttctcagac aatggaggtc taatagcacc aagtcgagtt agcaaattaa ctggaaggga 840

tttgggaatc caatcagaag cgttgataga caacagttgt gaatccaaat gcttttggag 900

agggggttct ataaatacaa agctcccttt tcaaaatctg tcacccagaa cagtaggtca 960

atgccccaaa tacgtaaatc agaggagttt actgcttgca acagggatga ggaatgtgcc 1020

agaagtggtg cagggaaggg gtctgtttgg tgcaatagca gggttcatag aaaacggatg 1080

ggaaggaatg gtagacggct ggtatggttt cagacaccaa aatgcccagg gcacaggcca 1140

agctgctgat tacaagagta ctcaagcagc tattgaccaa atcacaggga aactgaacag 1200

gttgattgag aagaccaaca ctgagtttga gtcaatagaa tctgaattca gtgagactga 1260

gcatcaaatt ggtaacgtca ttaattggac caaagattca ataaccgaca tttggactta 1320

caacgcagag ctattagtgg caatggagaa tcagcacaca attgacatgg ctgattcaga 1380

gatgctaaat ctgtatgaaa gggtaagaaa gcaactcaga cagaatgcag aagaagacgg 1440

aaagggatgt tttgagatat atcatacttg tgatgattcg tgcatggaga gtataaggaa 1500

caatacttat gaccattcac aatacagaga ggaggctctt ctgaatagac tgaacatcaa 1560

cccagtgaaa ctttcttcgg ggtacaaaga catcatactt tggtttagct tcggggaatc 1620

atgctttgtt cttctagccg ttgttatggg tcttgttttc ttctgcctga aaaatggaaa 1680

catgcgatgc acaatctgta tttagttaaa aacaccttgt ttctact 1727

<210> 127

<211> 1698

<212> DNA

<213> Artificial

<400> 127

atggagaaaa cactgctatt tgcagctatt ttcctttgtg tgaaagcaga tgagatctgt 60

atcgggtatt taagcaacaa ctcgacagac aaagttgaca caataattga gaacaatgtc 120

acggtcacta gctcagtgga actggttgag acagaacaca ctggatcatt ctgttcaatc 180

aatggaaaac aaccaataag ccttggagat tgttcatttg ctggatggat attaggaaac 240

cctatgtgtg atgaactaat tggaaagact tcatggtctt acattgtgga aaaacccaat 300

ccaacaaatg gaatctgtta cccaggaact ttagagagtg aagaagaact aagactgaaa 360

ttcagtggag ttttagaatt taacaaattc gaagtattca catcaaatgg atggggtgct 420

gtaaattcag gagtaggagt aaccgctgca tgcaaattcg ggggttctaa ttctttcttt 480

cgaaacatgg tatggctgat acaccaatca ggaacatatc ctgtaataaa gagaaccttt 540

aacaacacca aagggagaga tgtactgatt gtttggggaa ttcatcatcc tgctacactg 600

acagaacatc aagatctgta taaaaaggac agctcctatg tagcagtggg ttcagagacc 660

tacaacagaa gattcactcc agaaatcaac actaggccca gagtcaatgg acaggccgga 720

cggatgacat tctactggaa gatagtcaaa ccaggagaat caataacatt cgaatctaat 780

ggggcgttcc tagctcctag atatgctttt gagattgtct ctgttggaaa tgggaaactg 840

ttcaggagcg aactgaacat tgaatcatgc tctaccaaat gtcaaacaga aataggagga 900

attaatacga acaaaagctt ccacaatgtt cacagaaaca ctatcgggga ttgccccaag 960

tatgtgaatg tcaaatcctt aaagcttgca acaggaccta gaaatgtccc agcaatagca 1020

tcgagaggct tgtttggagc aatagctgga ttcatagaag ggggatggcc tggactgatc 1080

aatggatggt atgggttcca acacagggac gaagaaggaa caggcattgc agcagacaag 1140

gagtcaactc aaaaggcaat agaccagata acatccaagg taaataacat cgttgacagg 1200

atgaatacaa actttgagtc tgtgcaacac gaattcagtg aaatagagga aagaataaat 1260

caattatcaa aacacgtaga tgattctgtg gttgacatct ggtcatataa tgcacagctt 1320

ctcgttttac ttgaaaatga gaagacactg gacctccatg actcaaatgt caggaacctc 1380

catgagaaag tcagaagaat gctaaaggac aatgccaaag atgaggggaa cggatgcttc 1440

accttttacc ataagtgtga caataaatgc attgaacgag ttagaaacgg aacatatgat 1500

cataaagaat tcgaggagga atcaaaaatc aatcgccagg agattgaagg ggtgaaacta 1560

gattctagtg ggaatgtgta taaaatactg tcaatttaca gctgcattgc aagcagtctt 1620

gtattggcag cactcatcat ggggttcatg ttttgggcat gcagtaatgg atcatgtaga 1680

tgtaccattt gcatttag 1698

<210> 128

<211> 1695

<212> DNA

<213> Artificial

<400> 128

atggaaaaat tcatcatttt gagtactgtc ttggcagcaa gctttgcata tgacaaaatt 60

tgcattggat accaaacaaa caactcgact gaaacggtaa acacactaag tgaacaaaac 120

gttccggtga cgcaggtgga agaacttgta catcgtggga ttgatccgat cctgtgtgga 180

acggaactag gatcaccact agtgcttgat gactgttcat tagagggtct aatcctaggc 240

aatcccaaat gtgatcttta tttgaatggc agggaatggt catacatagt agagaggccc 300

aaagagatgg aaggagtttg ctatccaggg tcaattgaaa accaggaaga gctaagatct 360

ctgttttctt ccatcaaaaa atatgaaaga gtgaagatgt ttgatttcac caaatggaat 420

gtcacataca ctgggaccag caaggcctgc aataatacat caaaccaagg ctcattctat 480

aggagcatga gatggttgac cttaaaatca ggacaatttc cagtccaaac agatgagtac 540

aagaacacca gagattcaga cattgtattc acctgggcca ttcaccaccc accaacatct 600

gatgaacaag taaaattata caaaaatcct gatactctct cttcagtcac caccgtagaa 660

atcaatagga gcttcaagcc taatataggg ccaagaccac tcgtgagagg acaacaaggg 720

agaatggatt actactgggc tgttcttaaa cctggacaaa cagtcaaaat acaaaccaat 780

ggtaatctta ttgcacctga atatggtcac ttaatcacag ggaaatcaca tggcaggata 840

ctcaagaata atttgcccat gggacagtgt gtgactgaat gtcaattgaa cgagggtgta 900

atgaacacaa gcaaaccttt ccagaacact agtaagcact atattgggaa atgccccaaa 960

tacataccat cagggagttt aaaattggca atagggctca ggaatgtccc acaagttcaa 1020

gatcgggggc tctttggagc aattgcaggt ttcatagaag gcggatggcc agggctagtg 1080

gctggttggt acggatttca gcatcaaaat gcggagggga caggcatagc tgcagacaga 1140

gacagcaccc aaagggcaat agacaatatg caaaacaaac tcaacaatgt catcgacaaa 1200

atgaataaac aatttgaagt ggtgaatcat gagttttcag aagtggaaag cagaataaac 1260

atgattaatt ccaaaattga tgatcagata actgacatat gggcatacaa tgctgaattg 1320

cttgtcctat tggaaaatca gaagacatta gatgagcatg acgctaatgt aaggaatcta 1380

catgatcggg tcagaagagt cctgagggaa aatgcaattg acacaggaga cggctgcttt 1440

gagattttac ataaatgtga caacaattgt atggacacga ttagaaacgg gacatacaat 1500

cacaaagagt atgaggaaga aagcaaaatc gaacgacaga aagtcaatgg tgtgaaactt 1560

gaggagaatt ctacatataa aattctgagc atctacagca gtgttgcctc aagcttagtt 1620

ctactgctca tgattattgg gggtttcatt ttcgggtgtc aaaatggaaa tgttcgttgt 1680

actttctgta tttaa 1695

<210> 129

<211> 1768

<212> DNA

<213> Artificial

<400> 129

agcaaaagca ggggaaatat taacaatcag aaacaaacaa gatggctctc aatgtcattg 60

caactttgac acttataagt gtatgtgtac atgcagacag aatatgcgtg gggtatctga 120

gcaccaattc atcagaaagg gtcgacacgc tcctggaaaa tggggtccca gtcaccagct 180

ccattgatct gatcgagaca aaccacacag gaacatactg ttctctgaat ggagtcagtc 240

cagtgcattt gggagattgc agctttgaag gatggattgg tggaaaccca gcctgcacca 300

gcaactttgg gatcagagag tggtcatacc tgattgagga ccccgcggcc cctcatgggc 360

tttgctaccc tggagaatta aacaacaatg gtgaactcag acacttgttc agtggaatca 420

ggtcattcag tagaacggaa ttgatcccac ctacctcctg gggggaagta cttgacggta 480

caacatctgc ttgcagagat aacacgggaa ccaacagctt ctatcgaaat ttagtttggt 540

ttataaagaa gaataataga tatccagtta tcagtaagac ctacaacaat acaacgggaa 600

gggatgtttt agttttatgg ggaatacatc acccagtgtc tgtggatgag acaaagactc 660

tgtatgtcaa tagtgatcca tacacactgg tttccaccaa gtcttggagc gagaaatata 720

aactagaaac gggagtccga cctggctata atggacagag gagctggatg aaaatttatt 780

ggtctttgat acatccaggg gagatgatta ctttcgagag taatggtgga tttttagccc 840

caagatatgg gtacataatt gaagaatatg gaaaaggaag gattttccag agtcgcatca 900

gaatgtctag gtgcaacacc aagtgccaga cttcggttgg agggataaac acaaacagaa 960

cgttccaaaa catcgataag aatgctcttg gtgactgtcc caaatacata aagtctggcc 1020

aactcaagct agccactgga ctcagaaatg tgccagctat atcgaataga ggattgttcg 1080

gagcaattgc agggttcata gaaggaggct ggccaggttt aatcaatggt tggtacggtt 1140

ttcagcatca aaatgaacag ggaacaggaa tagctgcaga caaagaatca acacagaaag 1200

ctatagacca gataacaacc aaaataaata acattattga taaaatgaat gggaactatg 1260

attcaattag gggtgaattc aatcaagttg agaagcgtat aaacatgctt gcagacagaa 1320

tagatgatgc cgtgacggac atttggtcat acaatgccaa acttcttgta ttgctggaaa 1380

atgataaaac tttagatatg catgatgcta atgtaaagaa tttacatgag caagtacgaa 1440

gagaattgaa ggacaatgca attgacgaag gaaatggctg ttttgaactc cttcataaat 1500

gcaatgactc ctgcatggaa actataagaa atggaacgta tgaccacact gagtatgcag 1560

aggagtcaaa gttaaagagg caagaaatcg atgggatcaa actcaaatca gaagacaacg 1620

tttacaaagc attatcaata tacagttgca ttgcaagtag tgttgtacta gtaggactca 1680

tactctcttt catcatgtgg gcctgtagta gtgggaattg ccgattcaat gtttgtatat 1740

aagtagaaaa aacacccttg tttctact 1768

<210> 130

<211> 1749

<212> DNA

<213> Artificial

<400> 130

agcaaaagca ggggaaaatg attgcactca tattggttgc actggctctg agccacactg 60

cttattctca gatcacaaat gggacaacag gaaaccccat tatatgcttg gggcatcatg 120

cagtggaaaa cggcacatct gttaaaacac taacagacaa tcacgtagaa gttgtgtcag 180

ctaaagaatt agttgagacg aaccacactg atgaactgtg cccaagcccc ttgaagcttg 240

tcgacgggca agactgccac ctcatcaatg gtgcattggg gagtccaggc tgtgaccgtt 300

tgcaggacac cacttgggat gtcttcattg aaaggcccac tgcagtagac acatgttatc 360

cattcgacgt cccagattac cagagtctca gaagcatcct agcaagcagt gggagtttgg 420

agttcatcgc cgaacaattc acctggaatg gtgtcaaagt tgacggatca agcagtgctt 480

gtttgagggg cggtcgcaac agcttcttct cccgactaaa ctggctaacc aaagcaacaa 540

atggaaacta tggacctatt aacgtcacta aagaaaatac gggctcttat gtcaggctct 600

atctctgggg agtgcatcac ccatcaagcg ataatgagca aacggatctc tacaaggtgg 660

caacagggag agtaacagta tctacccgct cggaccaaat cagtattgtt cccaatatag 720

gaagtagacc gagggtaagg aatcagagcg gcaggataag catctactgg accctagtaa 780

acccagggga ctccatcatt ttcaacagta ttgggaattt gattgcacca agaggccact 840

acaaaataag caaatctact aagagcacag tgcttaaaag tgacaaaagg attgggtcat 900

gcacaagccc ttgcttaact gataaaggtt cgatccaaag tgacaaacct tttcagaatg 960

tatcaaggat tgctatagga aactgcccga aatatgtaaa gcaagggtcc ctgatgttag 1020

caactggaat gcgcaacatc cctggcaaac aggcaaaggg cttatttggg gcaattgctg 1080

gattcattga aaatggttgg caaggcctga ttgatgggtg gtatggattc aggcaccaaa 1140

atgctgaagg aacaggaact gctgcagacc tgaagtcaac tcaggcagcc attgatcaga 1200

taaatggcaa gctgaacaga ttgatagaga agacaaatga aaaatatcac caaatagaaa 1260

aggaattcga acaggtggaa ggaagaatac aagaccttga gaagtacgtt gaggacacta 1320

agattgattt gtggtcatac aatgctgaat tgctagtagc actagagaat cagcacacaa 1380

tagatgtcac agactccgaa atgaacaagc tttttgaaag agtaagaagg caattaagag 1440

agaatgcaga agatcaaggc aacggttgtt tcgagatatt ccatcagtgt gacaacaatt 1500

gtatagaaag cattagaaac ggaacttatg accacaacat ctacagggat gaagccatca 1560

acaatcgaat caaaataaat cctgtcactt tgacgatggg gtacaaggac ataatcctgt 1620

ggatttcttt ctccatgtca tgctttgtct tcgtggcact gattctggga tttgttctat 1680

gggcttgtca aaacgggaat atccgatgcc aaatctgtat ataaagaaaa aacacccttg 1740

tttctactc 1749

<210> 131

<211> 1762

<212> DNA

<213> Artificial

<400> 131

agcaaaagca ggggatacaa aatgaacact caaatcatcg tcattctagt cctcggactg 60

tcgatggtga gatctgacaa gatttgtctc gggcaccatg ccgtagcaaa tgggacaaaa 120

gtcaacacac taactgagaa aggagtggaa gtggtcaatg ccacggagac agtggagatt 180

acaggaataa ataaagtgtg cacaaaaggg aagaaagcgg tggacttggg atcttgtgga 240

atactgggaa ctatcattgg gcctccacaa tgtgactctc atcttaaatt caaagctgat 300

ctgataatag aaagaagaaa ttcaagtgac atctgttacc cagggaaatt cactaatgag 360

gaagcactga gacaaataat cagagaatct ggtggaattg acaaagagcc aatgggattt 420

agatattcag gaataaaaac agacggggca accagtgcgt gtaagagaac agtgtcctct 480

ttctactcag aaatgaaatg gcttttatcc agcaaggcta accaggtgtt cccacaactg 540

aatcagacat acaggaacaa cagaaaagaa ccagccctaa ttgtttgggg agtacatcat 600

tcaagttcct tggatgagca aaataagcta tatggagctg ggaacaagct gataacagta 660

ggaagctcaa aataccaaca atcgttttca ccaagtccag gggacaggcc caaagtgaat 720

ggtcaggccg ggaggatcga ctttcattgg atgctattgg acccagggga tacagtcact 780

tttaccttca atggtgcatt catagcccca gatagagcca cctttctccg ctctaatgcc 840

ccatcgggag ttgagtacaa tgggaagtca ctgggaatac agagtgatgc acaaattgat 900

gaatcatgtg aaggggaatg cttctacagt ggagggacaa taaacagccc tttgccattt 960

caaaacatcg atagttgggc tgtcggaagg tgccccagat atgtaaagca atcaagcctg 1020

ccgctggcct taggaatgaa aaatgtacca gagaaaatac atactagggg actgttcggt 1080

gcaattgcag gattcatcga gaatggatgg gaaggactca ttgatggatg gtatggattt 1140

aggcatcaaa atgcacaggg gcagggaaca gctgctgact acaagagtac tcaggctgca 1200

attgaccaga taacagggaa acttaataga ttaattgaaa aaaccaacac acagtttgaa 1260

ctcatagaca atgagttcac tgaagtggag cagcagatag gcaatgtaat aaactggaca 1320

agggactcct tgactgagat ctggtcatac aatgctgaac ttctagtagc aatggaaaat 1380

cagcatacaa ttgaccttgc agattctgaa atgaacaaac tctatgagag agtgagaaga 1440

cagctaaggg agaatgccga ggaggatgga actggatgtt ttgagatttt ccaccgatgt 1500

gacgatcaat gtatggagag catacgaaat aatacttaca atcacactga atatcgacag 1560

gaagccttac agaataggat aatgatcaat ccggtaaagc ttagtggtgg gtacaaagat 1620

gtgatactat ggtttagctt cggggcatca tgtgtaatgc ttctagccat tgctatgggt 1680

cttattttca tgtgtgtgaa aaacgggaat ctgcggtgca ctatctgtat ataattattt 1740

gaaaaacacc cttgtttcta ct 1762

<210> 132

<211> 1730

<212> DNA

<213> Artificial

<400> 132

ggacattgtc aaaacagcag aaatgatggt caaggtgctc taccttctca tcatagtgtt 60

gggcagatac tcaaaagcag acaaaatatg cataggatac ctgagcaata actcaacaga 120

taaagtagat acactgacag aaaatggagt tcctgtgacc agctcagtgg accttgtaga 180

aacaaaccat actggaacat actgctcatt gaatgggatc agcccggtcc atcttggtga 240

ctgcagtttt gagggatgga ttgtagggaa tccctcctgt gccaccaaca tcaacataag 300

agagtggtcg tatctaatcg aagatcctaa tgctcctaac aagctctgct tcccaggaga 360

gttggataac aatggtgagc tgcggcacct tttcagtgga gtaaactctt ttagcaggac 420

agaattgata agccccagca aatggggggc tgttctggat ggagtcactg cttcatgtct 480

cgacagaggg gcaagcagtt tttacagaaa tctggtctgg ctggtaaaac agaatgagag 540

atatcccgtt gtaagagggg attataacaa cacaacagga cgagatgttt tggtgatttg 600

gggaatccac catccagatt cggaagcaac agccacaaaa ctatatgtca acaaaaaccc 660

ctacacattg gtatcgacaa aagagtggag caaacgctat gagctcgaga tcgggaccag 720

aataggtgac ggacaaaaga gttggatgaa aatatattgg cacctcatgc gccccggcga 780

aaggataacg tttgagagca atggaggcct tctagccccc agatacggat atatcattga 840

gaagtacggt tcaggaagaa ttttccagag tggagtgagg atggcaaaat gcaacacaaa 900

gtgccaaacg tccgtaggtg gaataaacac caataaaact ttccagaaca tagacaggaa 960

tgccctggga gaatgtccga agtacataaa gtctgggcaa ttaaaacttg ctactggcct 1020

gagaaatgtc ccatccatta atggaagagg tctgtttggt gcaattgcag gattcataga 1080

gggagggtgg cctggtctga tcaatgggtg gtatggtttc cagcatcaga atgaacaagg 1140

gactggcatt gctgcagata aagcttccac ccagaaagca ataaatgaaa taacaacgaa 1200

aattaacaac ataatagaaa agatgaacgg aaattatgat tcaataagag gagaattcaa 1260

tcaagtagaa aagagaatca acatgctggc tgatcgagtt gatgatgcag taaccgatgt 1320

ttggtcatac aatgctaaac ttcttgtact gcttgaaaat ggtagaacat tagacttgca 1380

cgatgcaaat gtcagaaact tacatgatca ggtcaaaagg gtgttgaaag acaatgcaat 1440

tgacgaagga aatggttgct tcaatcttct ccacaagtgc aacgactcat gcatggaaac 1500

cattagaaat ggcacctaca atcatgaaga ttacaaggaa gaatcacaac tgaatagaca 1560

ggaaattgag gggataaaac tgaaaactga agacaatgtt tataaaatac tatcgattta 1620

tagctgcatt gcaagcagta ctgtactggt gggtctcata cttgcattta taatgtgggc 1680

atgcagcagt ggtaattgcc ggttcaatgt gtgtatataa tcggaaaaaa 1730

<210> 133

<211> 1784

<212> DNA

<213> Artificial

<400> 133

agcagaagca gggtcactat tactctgtgc tactatggag ctgattatcc tactaatcct 60

tctcaatccc tatacttttg tgttagggga cagaatatgc ataggctatc aagcaaacca 120

aaataaccaa acggttaaca ctctgctaga acagaatgtt ccagttaccg gagcgcagga 180

aatactagaa accaatcaca acggaaaatt atgcagccta aatggagtcc caccattgga 240

cctacaatca tgcactctag ccgggtggtt attggggaat ccgaactgtg acaacctatt 300

ggaagcggaa gaatggtcgt atataaaaat aaatgaaaat gcccctgacg atctttgctt 360

ccctggaaac ttcgaaaact tacaggactt actactagaa atgtcaggag ttcaaaattt 420

caccaaggtg aaactattca acccccaaag tatgactggg gtgactacca ataatgtcga 480

ccagacttgc ccttttgaag ggaaaccatc tttctacaga aacctcaact ggatacaagg 540

gaatagcggt ttacctttca atatagaaat caagaatcca actagcaatc cattgctcct 600

tctctggggg atccacaaca ccaaggatgc agcacaacaa agaaatctct atgggaatga 660

ttactcttat actattttta actttgggga gaaaagtgaa gaatttcgac ctgacatcgg 720

gcaaagagat gaaatcaaag cccatcaaga cagaatcgat tactactggg gaagcctacc 780

agcgcagagc actctaagga tagaatcaac cggaaatcta atcgcaccag aatatggctt 840

ttattacaag cgaaaagagg ggaaaggggg gctaatgaaa agtaagctgc caattagtga 900

ctgctcaacc aaatgccaaa cacccctagg agctctcaac agcaccctgc cctttcaaaa 960

cgtccatcaa caaacaattg gtaattgtcc caaatatgtg aaagctacct ccttgatgct 1020

tgccacaggg ctaagaaaca acccccaaat ggagggaaga ggtctctttg gagccattgc 1080

cgggttcatt gaaggagggt ggcaagggat gattgatggt tggtatggat accaccacga 1140

gaatcaagaa ggaagcggtt acgctgctga caaagaagcc acccaaaagg ctgtcgatgc 1200

cataacgaac aaagttaaca gtattattga caaaatgaac agccaatttg aatccaacat 1260

caaagagttc aacaggttgg aactcaggat acaacaccta agtgatagag ttgatgatgc 1320

gttacttgac atttggtcct ataatactga attacttgtc ctcttagaaa atgaaagaac 1380

ccttgatttc catgatgcta acgtaaagaa cctctttgaa aaagtgaagg cccaattgaa 1440

ggacaatgca atagatgagg gaaatggttg ctttctactt ctacacaagt gtaacaactc 1500

ttgcatggac gacataaaga atgggacata caaatacatg gactataggg aagagtcaca 1560

catcgaaaaa caaaaaatcg atggagtaaa actaactgat tactctagat actataccat 1620

gactctgtac tctaccattg catcatccgt cgtgcttggc tcattgataa tagccgcttt 1680

tctttggggg tgccaaaaag gctcaatcca atgtaaaata tgcatataga gcggtgggat 1740

tagccttgtc attcagaaaa acaaaaaaga cccttgtttc tact 1784

<210> 134

<211> 1771

<212> DNA

<213> Artificial

<400> 134

agcagaagca gggtgattat tattcagaat gattacaata cttatcttgg tactccctat 60

tgttgtaggt gaccaaatat gcattggcta tcattcaaat aattcaacac aaacagtgaa 120

tactctcctt gaatcaaatg taccagtgac ttcctctcac agcatcctag aaaaagaaca 180

caatggtttg ctttgcaagc taaaagggaa agcacccttg gaccttattg actgctctct 240

tcctgcatgg cttatgggaa acccaaaatg tgacgaactc ttaacagcaa gcgaatgggc 300

ctacataaaa gaagacccag aacctgaaaa tggaatctgt tttccaggag attttgattc 360

tttagaggat ctgattttat tggtttctaa cactgaccat ttcagaaaag agaaaataat 420

agacatgacc agattctctg atgtgactac aaacaacgta gacagtgcat gcccatatga 480

cacaaatggt gcttcctttt acagaaatct aaactgggtg cagcaaaaca aaggcaagca 540

actgattttt cactaccaga attctgaaaa caacccactt ttgataattt ggggagtaca 600

ccagacatct aatgctgcag aacaaaacac atactatggc tcacagactg gctcaacaac 660

catcactatt ggggaagaaa caaacactta tccactagtg ataagtgaaa gttctattct 720

taacggtcac tctgatagaa taaattactt ttggggagtt gtcaatccta atcagaattt 780

ttcaattgtc agtacaggga atttcatctg gccagagtac ggatactttt tccaaaaaac 840

aaccaatata agtggaataa taaaatcaag tgaaaagata agtgattgtg acacaatctg 900

ccagacaaaa attggggcaa taaacagcac actgcctttt cagaatatcc atcaaaatgc 960

gattggagat tgtcctaaat atgtgaaagc ccaagaactt gttcttgcaa ctggattaag 1020

gaacaatcca ataaaagaaa caagagggct ttttggtgca attgcaggtt tcatcgaggg 1080

aggatggcaa ggattgattg atggttggta tgggtatcac caccagaact cagaaggttc 1140

aggctatgct gctgacaaag aagcaaccca gaaggctgtt gatgcaataa ccacaaaagt 1200

aaacaacata atagacaaaa tgaacacgca atttgaatca actgccaaag aattcaacaa 1260

aattgaaatg agaataaaac atctcagtga cagagttgat gatggcttct tggatgtttg 1320

gagttacaat gctgaattac tcgttttgct ggaaaatgaa agaactctgg acttccatga 1380

tgcaaatgtt aacaatttgt atcaaaaagt gaaagtccag ctgaaagaca atgcaattga 1440

catgggaaac ggctgtttca agattctaca caaatgcaac aacacatgta tggatgacat 1500

taaaaacgga acatacaatt attatgaata cagaaaggaa agccacttgg agaaacaaaa 1560

aattgacggt gtgaagctat cagaaaacag ctcatataaa ataatgatca tttactcaac 1620

agtggcaagt tcagtagtgc ttggcttgat tatactagcc gcaattgaat ggggctgttt 1680

taaagggaac ctgcaatgca gaatatgtat ttgaggctgt ggtgttagct aatgtcaatc 1740

tattattgca aaaaacaccc ttgtttctac t 1771

<210> 135

<211> 1080

<212> DNA

<213> Artificial

<400> 135

ataatctcaa tatgggttag tcattcaatc caaactggaa gcaagagcca ccctgaaaca 60

tgcaatcaaa gtgtcattac ctacgaaaac aatacgtggg tgaatcaaac ataccttaac 120

ataagtaata ccaatttaat tgcagaacag actgtagctc cagtaacact agcaggcaat 180

tcctctctct gtcccatcag tggatgggct atatacagca aggataatgg tataagaata 240

ggttccaaag gagatgtatt tgtcatcaga gagcctttta tttcatgctc gcacttggaa 300

tgcagaactt tctttctaac tcaaggggcc ttgttgaatg acaagcattc caatggaacc 360

gttaaagaca gaagccctta tagaacccta atgagctgtc ctgttggtga agctccctct 420

ccatacaatt caaggtttga gtctgtcgct tggtcggcaa gtgcttgcca cgatggcatt 480

agctggttga caattggtat ttccggccca gacaatgggg cagtggctgt attgaagtac 540

aatggcataa taacagatac tatcaagagt tggagaaaca acatattgag gacacaagaa 600

tctgaatgtg cctgcattaa tggttcttgc tttaccataa tgactgatgg accaagtaat 660

ggtcaggctt catacaagat tttcaaaata gaaaagggaa aggtagtaaa atcagttgag 720

ttgaatgccc ctaattacca ctatgaagag tgttcctgtt atcctgatgc tagtgaggtg 780

atgtgtgtat gcagggacaa ttggcatggt tcaaatcggc catgggtgtt cttcaatcag 840

aatctagagt atcaaatagg atatatatgc agcggagttt ttggagacaa tccacgcccc 900

aatgatggga caggcagttg tggtccagta tcttctaatg gggcatatgg ggtaaaaggg 960

ttttcgttta gatacggtaa cggtgtttgg ataggaagaa ctaaaagtac tagctcaagg 1020

agtgggtttg agatgatttg ggaccccaat ggatggacag agacggacaa cagtttctct 1080

<210> 136

<211> 1351

<212> DNA

<213> Artificial

<400> 136

accattgcaa cagtatgttt cctcatgcaa attgctatcc tggcaacgac ggtgacattg 60

cacttcaagc aaaatgaatg caacatccct tcgaataatc aagtagtgcc atgtgaccca 120

atcataatag aaagaaacat aacagagata gtgtatttga ataatactgt catagaaaag 180

gaaatttgtc ctaaagcggt agaatacagg aattggtcaa aaccacagtg tcagattgca 240

gggtttgctc ctttctccaa ggacaactca atccggcttt ctgctggtgg agacatttgg 300

gtgacaagag aaccttatgt gtcatgcagc cctgataaat gttatcaatt tgcacttgga 360

cagggaacca cgctagacaa caaacattca aatggtacaa tacatgatag aattcctcat 420

cgaacccttt taatgagcga gttgggtgtt ccattccatt taggaaccaa acaagtgtgc 480

atagcatggt ccagttcaag ctgccatgat gggaaagcat ggttacatgt ttgtgtcact 540

ggagacgaca gaaatgcaac agctagtttc atttacaatg ggatgcttgt tgatagtatt 600

ggttcatggt ctcagaacat cctcagaact caggagtcag agtgtgtttg cattaatgga 660

acttgtacag tagtaatgac tgatggaagt gcgtcaggaa gggctgatac tagaatactg 720

ttcgttatag aggggagaat tattcatatc agcccattat caggaagtgc tcagcatata 780

gaggagtgct cctgctatcc tcgataccca gacgttagat gtgtttgcag agacaattgg 840

aagggctcta ataggcccat tatagatata aatatggcag attatagcat tgattccaat 900

tatgtgtgct caggacttgt tggcgataca ccaaggaacg acgatagctc tagcagcagc 960

aattgcaagg atcccaataa cgagagagga agcccgggag tgaaagggtg ggcctttgac 1020

tatggaaatg acgtttggat gggaagaaca atcaacaagg attcacgctc agggtatgag 1080

actttcagag tcattggtgg ttgggccaca gctaattcca aatcccaggt aaatagacaa 1140

gtcatagttg acaatagcaa ctggtctggt tactctggta ttttctctgt tgaaggcaaa 1200

cactgcatca ataggtgttt ttatgtggag ttgataagag ggaggccaca ggagactaga 1260

gtatggtgga cctcaaatag tatcgttgtg ttctgtggca cttcgggtac ctatggaaca 1320

ggctcatggc ctgatggggc gaatatcaat t 1351

<210> 137

<211> 1410

<212> DNA

<213> Artificial

<400> 137

atgaatccaa atcagaagat aataacaatt ggtgtagtga acactactct atcaacaata 60

gcccttctca ttggagttgg aaacctgatt ttcaacaccg ttatacatga gaaaataggg 120

gaccatcaaa ctgtggtgta tccaacaata acagccccgg tagtaccaaa ctgcagtgac 180

accataatta catacaacaa cactgtgata aacaacataa caacaacaat aataactgaa 240

gcagaaaggc atttcaagcc ctcactgccg ctgtgcccct tcagaggatt tttccctttt 300

cataaggaca atgcaatacg attgggtgag aacaaagacg taatagtcac aagagagcct 360

tacgttagtt gcgacaatga taactgctgg tcctttgctc ttgcccaagg ggcattactg 420

ggaactaaac acagcaatgg aaccatcaaa gacagaacac cctatagatc actgattcga 480

tttccaatag gaacagcccc agtactaggg aattacaaag agatatgtgt agcttggtcg 540

agtagcagtt gcttcgatgg gaaggagtgg atgcatgttt gcatgacagg gaacgacaat 600

gatgcaagtg cccaaataat atatgcaggg aaaatgacag actccattaa atcatggaga 660

agggacatac taagaaccca agaatctgaa tgtcaatgta ttgatgggac ctgtgttgtc 720

gctgttacag atggccctgc ggctaatagt gcagaccata gagtctattg gatacgagaa 780

ggaaaggtaa taaagtatga aaacgtcccc aaaacaaaga tacaacactt ggaggagtgt 840

tcctgttatg tggacactga cgtgtactgc gtgtgtaggg acaattggaa aggttccaac 900

aggccttgga tgaggatcaa caatgagact atactagaaa cagggtatgt atgtagcaaa 960

ttccactcag acactcccag accagccgac ccttcaacag tatcatgtga ttccccaagt 1020

aatgtcaatg gaggaccagg agttaaaggg tttggcttca agacgggcga tgatgtatgg 1080

ttggggagga ctgtatcgac taatgggagg tcaggctttg agatcatcaa agtcacagag 1140

gggtggatca attctcccaa tcatgccaaa tcagttacac aaacattagt gtcaaacaat 1200

gattggtcgg gttactcagg gagcttcatt gttgaaaaca acggctgttt ccagccttgc 1260

ttctatattg aacttatacg agggaggacc aataagaatg atgacgtttc ttggacgagc 1320

aatagtatag ttactttctg tggactagac aatgaacctg gatcgggaaa ttggcctgat 1380

ggttccaaca ttgggtttat gcccaagtaa 1410

<210> 138

<211> 1020

<212> DNA

<213> Artificial

<400> 138

atgaatccaa atcagaaaat cataaccatc ggtagtgcga gtattgtatt aacaacaata 60

gggttgctcc ttcaaataac aagcttatgc tcaatatggt ttagccatta caaccaggtg 120

acccaaacac atgaacaacc ctgttcgaac aacacaacga attactataa tgaaaccttt 180

gtcaatgtca ccaatgtgca gaacaactat accacagtaa ttgaaccctc agcccctgat 240

gtggtccatt actctagtgg aagagacttg tgcccaataa aagggtgggc acctctaagt 300

aaagacaatg gaattagaat tggatcccga ggcgaggtat ttgtcatacg agagcctttc 360

atatcatgtt ccattaatga atgcaggacc tttttcttga ctcagggagc cctcctcaat 420

gacaagcatt cgaacggaac agtaaaagac agaagtccct ttcgtacgtt aatgagttgt 480

ccagtaggag ttgccccctc tcctagcaac agccgttttg aatctgtggc atggtctgcc 540

acagcatgca gtgatggacc cggttggcta acattaggga tcactggccc agatgctact 600

gctgtagcag tactgaaata caatggtata ataacagaca cgttaaaaag ctggaagggt 660

aatatcatgc gaacacaaga gtctgaatgc gtatgccagg atgagttttg ctatacctta 720

ataacagacg gaccgtctga cgcacaagct ttctataaga tactaaagat caggaaaggg 780

aagatagtaa gtgtgaagga tgtggatgca acagggttcc attttgaaga atgttcctgt 840

tacccgagcg ggacagatgt tgaatgtgtc tgtagagaca actggcgggg gagcaatagg 900

ccatggataa gattcaacag tgatcttgat tatcaaattg gctatgtatg tagcgggata 960

tttggggaca atcccaggcc tgtggatggc ataggctcat gtaacagtcc agtcaacaac 1020

<210> 139

<211> 1020

<212> DNA

<213> Artificial

<400> 139

atgaacccaa atcagaaaat aataacaatt ggttctgtgt cattggcact agttgtattc 60

aacatactgc ttcatattgc atcaatagtc ataggaataa tatcagtgac gaaagaaatc 120

agtgtatcat caacctgcaa taccactgag gtttacaatg aaactgtaag gctggaaact 180

atcacaattc ctatcaacaa cactgtttac atagaaagag agtcacatca agaacctgat 240

tttttgaaca atacagaacc tctctgcaat gtgtccgggt tcgcaatagt ttccaaggac 300

aatggaatca gaattgggtc aagaggacat gtgtttgtca taagagaacc attcgtggca 360

tgtggtccca cagaatgtag gacttttttc ctaactcaag gtgccctatt gaacgataaa 420

cactccaaca atacagtaaa agataggagt ccttaccgtg cattgatgag tgtgccatta 480

ggatcttcac ccaatgctta tcaggccaag tttgagtctg ttgcatggtc ggccacagca 540

tgccatgatg gcaaagggtg gctggcagta gggataagtg gtgcagatga cgatgcttat 600

gctgtaatcc attatggggg gatgccaaca gatgtggtga ggtcatggag aaagcaaatt 660

ctaagaacac aggaatcgtc atgtgtgtgt atgaaaggaa actgttattg ggtaatgacg 720

gatggtcctg caaacagtca ggctagttac aagattttca agtctcacaa ggggatggtg 780

acaaatgaaa gagaagtgtc atttcaggga ggtcacattg aagagtgctc ttgctatccc 840

aatttgggta aagtggagtg tgtttgccga gataattgga atggaatgaa tagaccagtt 900

ttgacttttg atgaggactt gaattatgag gtgggttatt tatgtgccgg agtaccgaca 960

gacactccac gggtccaaga caataggttc actggttcct gtactaatgc tgttggaggg 1020

<210> 140

<211> 993

<212> DNA

<213> Artificial

<400> 140

agagggcgac atgcgaatgg gaccatacat gataggagcc cattcagagc tctcataagc 60

tgggaaatgg gtcaggcacc cagtccatat aatgttaggg tcgagtgcat aggatggtca 120

agcacgtcat gccacgatgg catatcaagg atgtcaatat gcatgtcggg agcgaacaac 180

aatgcatcag cagtggtgtg gtacgggggg aggccagtaa cagaaattcc atcatgggcc 240

ggaaatattc ttagaaccca agaatcagaa tgtgtgtgcc ataaggggat ctgcccagta 300

gtcatgacag atggcccagc aaacaatagg gcagcaacta agataatcta tttcaaagag 360

ggaaagatac agaaaatcga agaactggca gggaatactc agcacatcga agaatgttca 420

tgctatggag cagtaggggt gatcaaatgc atatgcaggg acaattggaa gggggcaaat 480

agaccggtaa taactataga tcccgaaatg atgacccaca caagcaagta cttgtgctca 540

aagatcctaa ccgatacaag ccgccccaat gatcccacca atgggaactg tgatgcgcca 600

ataacagggg ggagcccaga tcctggggtg aaggggtttg cattcctcga cagggagaac 660

tcatggctcg gaaggacaat tagcaaagac tccagatcag gctatgaaat gttaaaagtc 720

ccaaatgcag aaaccgacac ccaatcgggg ccaatctcac accaggtgat tgtcaacaac 780

caaaattggt caggatactc aggggcattc atagactact gggcaaacaa agagtgcttc 840

aatccttgtt tttatgtgga attaatcaga gggagaccca aagagagtag tgtactgtgg 900

acttcaaata gcattgtagc tctctgtgga tccaaagagc gattgggatc atggtcctgg 960

catgatggtg ctgaaatcat ctactttaag tag 993

<210> 141

<211> 1113

<212> DNA

<213> Artificial

<400> 141

gacaatgcca taagattcgg tgaaagtgaa caaataatag tgacaagaga gccgtatgtg 60

tcatgtgatc cattaggatg taagatgtac gcactgcatc aaggaacaac cattagaaac 120

aagcattcaa acggaacaat acacgacagg actgctttca gaggcttgat atcaactcct 180

ttggggagcc cccctgtagt cagcaatagt gactttcttt gtgtagggtg gtcaagcacc 240

agttgccatg acggcatcgg gcggatgacc atttgtgtgc agggaaataa tgacaacgca 300

acagctacag tgtactatga ccgaaggctc actaccacaa taaaaacatg ggcaggaaac 360

atccttagga cgcaagaatc ggaatgtgtg tgccacaatg ggacatgtgt agtaataatg 420

accgatggat cggcaagcag ccaggcatat acaaaagttc tgtatttcca caaaggacta 480

gtaataaaag aggaagccct taaaggatca gccagacaca tagaggagtg ctcatgctat 540

gggcacaatt cgaaggtgac ttgtgtatgc agggacaact ggcaaggggc caatagacca 600

gtgattgaaa tagatatgaa tgccatggag catacaagtc aatatctatg tacaggagtt 660

ctcactgaca cgagcagacc atcagataaa tcaatgggag actgtaataa tccaatcact 720

gggagcccgg gagcccctgg ggtcaaagga ttcggcttcc tggatagtag caatacatgg 780

ttgggccgca caataagtcc tcgttccagg agtggttttg agatgttgaa gatacctaat 840

gctgagacag acccaaattc taaaatcacc gagaggcaag aaatagttga caacaacaat 900

tggtcaggat actcaggaag tttcattgac tattgggatg aaagcagtga gtgttacaac 960

ccctgttttt atgttgaatt aataagaggg aggcctgaag aagccaagta tgttgggtgg 1020

acgagcaaca gtttaattgc actatgtgga agcccaatct cagttgggtc cggttccttc 1080

cccgatgggg cacaaatcca atacttttcg taa 1113

<210> 142

<211> 1113

<212> DNA

<213> Artificial

<400> 142

gacaatggaa taagaattgg atcgagaggt catgtttttg ttataaggga accatttgtt 60

tcttgttctc caacagagtg caggacgttc ttccttaccc agggttcctt actcaatgac 120

aaacactcca atggcacagt gaaagaccgg agcccctata gaactttaat gagtgtggaa 180

atagggcaat cacccaatgt gtaccaggca agatttgagg cagtggcatg gtcagccact 240

gcatgccacg atgggaagaa atggatgaca attggagtaa cgggccctga tgccaaagca 300

gtagcagtgg tgcattatgg gggaattccc actgatgtaa tcaattcctg ggcaggagac 360

attctaagaa ctcaggaatc atcatgcact tgcattcaag gtgaatgtta ttgggtaatg 420

acggatggac cagcaaacag acaagcacaa tacagagcat tcaaggccaa gcaggggaaa 480

atagttgggc aaactgagat cagtttcaat ggaagccata tagaagaatg ctcatgctac 540

cctaatgaag gtaaagtgga atgtgtttgt agggacaact ggactggaac caacagacca 600

gtattggtga tttctccaga tttgtcttac agagccggat acttgtgtgc aggtctcccc 660

agtgacactc caagaggaga agatagtcaa ttcacaggat cctgcactag cccggtggga 720

aatcaggggt atggagttaa gggatttgga ttcaggcagg gcaatgatgt atggatggga 780

aggaccatta gcaggacatc aagatcaggg tttgaaatcc tgaaggtcag aaatggctgg 840

gtacaaaata gtaaagagca gatcaaaagg caagttgtgg tcgataattt gaaatggtca 900

ggatacagtg gttccttcac actaccggtg gaattaacaa aaagaaattg tctggtccca 960

tgcttttggg ttgagatgat aagggggaag ccagaagaaa agactatatg gacctcaagt 1020

agctccattg tgatgtgtgg agtagaccat gagattgccg actggtcatg gcacgatgga 1080

gctattcttc cttttgacat cgataagatg taa 1113

<210> 143

<211> 1067

<212> DNA

<213> Artificial

<400> 143

gacccagatg aatgcaggtt ctatgctctc agccaaggaa caacaatcag agggaaacac 60

tcaaacggaa caatacacga taggtcccag tatcgcgccc tgataagctg gccactatca 120

tcaccgccca cagtgtacaa cagcagggtg gaatgcattg ggtggtcaag tactagttgc 180

catgatggca aatccaggat gtcaatatgt atatcaggac caaacaacaa tgcatctgca 240

gtagtatggt acaacagaag gcctgttgca gaaattaaca catgggcccg aaacatacta 300

agaacacagg aatctgaatg tgtatgccac aacggcgtat gcccagtagt gttcaccgat 360

gggtctgcca ctggacctgc agacacaaga atatactatt ttaaagaggg gaaaatattg 420

aaatgggagt ctctgactgg aactgctaag catattgaag aatgctcatg ttacgggaag 480

cgaacaggaa ttacctgcac atgcagggac aattggcagg gctcaaatag accagtgatt 540

cagatagacc cagtagcaat gacacacact agtcaatata tatgcagtcc tgttcttaca 600

gacaatcccc gaccgaatga cccaaatata ggtaagtgta atgaccctta tccaggtaat 660

aataacaatg gagtcaaggg attctcatac ctggatgggg ctaacacttg gctagggagg 720

acaataagca cagcctcgag gtctggatac gagatgttaa aagtgccaaa tgcattgaca 780

gatgatagat caaagcccat tcaaggtcag acaattgtat taaacgctga ctggagtggt 840

tacagtggat ctttcatgga ctattgggct gaaggggact gctatcgagc gtgtttttat 900

gtggagttga tacgtggaag acccaaggag gataaagtgt ggtggaccag caatagtata 960

gtatcgatgt gttccagtac agaattcctg ggacaatgga actggcctga tggggctaaa 1020

atagagtact tcctctaaga tgaagtaaaa agacccttgt ttctact 1067

<210> 144

<211> 1030

<212> DNA

<213> Artificial

<400> 144

cgtatgttag ttgtgacttg acttcctgtt tcaaattctt tattgcttac ggccttagtg 60

caaatcagca cttattgaac acaagtatgg aatgggagga gagcctatac aaaactccaa 120

ttggaagcgc aaatacacta agcacttcag aaatgatcct cccggggagg agttcatcag 180

catgcttcga cgggttaaaa tggaccgtct tggtgtctaa tggcagagac cggaacagct 240

tcataatgat caaatatgga gaggaaataa cagacacttt ctcggccagc agaggaggtc 300

ccctgcgact tcccaattca gaatgcatct gtgtagaagg aagttgcttt gtactggtaa 360

gtgacgggcc taacgtgaat caaagtgtcc accggatcta tgaactccaa aacggaacag 420

tccagaggtg gaagcagcta aatacgactg gcataaactt tgaatacagc acgtgctata 480

caatcaacaa cctgataaag tgcactggaa caaatctctg gaatgatgcc aaaaggcctt 540

tgctccgatt cactaaggac ctcaactatc agatcgtaga gccctgcaat ggagctccta 600

cagatttccc cagaggcggg ctgaccaccc caagttgcaa gatggctcag gaaaaaggag 660

aaggagggat ccaaggtttc atacttgacg agaaaccagc ctggacctca aaaacgaaga 720

ctgagttatc tcagaatggg tttgtattag aacaaatccc tgacgggata gaaagtgagg 780

gaacagtttc attaagctat gaactttttt ctaataagag aaccggaaga agcggatttt 840

ttcaacccaa aggagatctc atttctgaat gccaacgagt ctgtttctgg ctggaaatag 900

aagaccaaac agtaggccta ggaatgattc aagagctcag caccttttgt ggaataaact 960

cacctgttca gaatataaat tgggattcat gaccaatgga caatgaatga aaaaactcct 1020

tgtttctact 1030

<210> 145

<211> 946

<212> DNA

<213> Artificial

<400> 145

aacaaatgaa tgtttacagt gtaaagttag gagaccctcc aacacctgac aagttaaaat 60

ttgaagctgt tggctggagt gccagctcgt gtcatgatgg ctttcagtgg actgtcctgt 120

ccgttgcagg agacggtttt gtgagcatcc tttatggagg aattataact gatacaattc 180

atccaacaaa tggaggccca ctgagaacac aagcttcatc ttgcatatgc aatgatggaa 240

cttgttatac aatcattgct gatggaacca cttacactgc atcttctcac agactttaca 300

gactagtcaa tggaacatct gccggctgga aggcccttga taccacaggg ttcaattttg 360

agtttccgac ttgctactat acaagtggca aagtaaaatg taccggaaca aatctttgga 420

atgatgccaa gaggcccttt cttgaatttg accagtcctt cacttacact ttcaaggagc 480

catgcttggg gttccttggg gacaccccaa gagggattga caccactaat tactgtgaca 540

agacaacaac agagggagag ggtggaatcc aaggtttcat gattgaaggc tcaaactcct 600

ggataggaag aattattaat ccaggatcca agaaaggatt tgaaatttat aagttcctgg 660

gaacattgtt ttctgtccaa actgtaggaa ataggaacta ccaattgtta agtaacagca 720

caattgggag atcaggcctg tatcagcctg cttatgaatc acgtgattgt caagagttgt 780

gtttttggat tgaaattgct gcaactacca aagcaggctt gtcatccaat gatctgatta 840

ctttttgtgg gacaggaggc tcaatgccag atgtcaactg ggggtaagta tatgattaca 900

ttcatatttt aaatggatgt ataagaaaaa actccttgtt tctact 946

Claims

1. a kind of LAMP primer group for being used to expand influenza A virus, it is characterised in that the LAMP primer group includes following One group in 29 groups of primer sets or several groups：

For the primer sets of IAV H1 areas fragment amplification, the primer sets include two pairs of primers, are respectively：H1-F3：Its nucleosides Sour nucleotide sequence is as shown in SEQ ID NO.1, H1-B3：Its nucleotide sequence is as shown in SEQ ID NO.2；And H1-FIP： Its nucleotide sequence is as shown in SEQ ID NO.3, H1-BIP：Its nucleotide sequence is as shown in SEQ ID NO.4；

For the primer sets of IAV H2 areas fragment amplification, the primer sets include two pairs of primers, are respectively：H2-F3：Its nucleosides Acid sequence is as shown in SEQ ID NO.5, H2-B3：Its nucleotide sequence is as shown in SEQ ID NO.6；And H2-FIP：Its core Nucleotide sequence is as shown in SEQ ID NO.7, H2-BIP：Its nucleotide sequence is as shown in SEQ ID NO.8；

For the primer sets of IAV H3 areas fragment amplification, the primer sets include two pairs of primers, are respectively：H3-F3：Its nucleosides Acid sequence is as shown in SEQ ID NO.9, H3-B3：Its nucleotide sequence is as shown in SEQ ID NO.10；And H3-FIP：Its core Nucleotide sequence is as shown in SEQ ID NO.11, H3-BIP：Its nucleotide sequence is as shown in SEQ ID NO.12；

For the primer sets of IAV H4 areas fragment amplification, the primer sets include two pairs of primers, are respectively：H4-F3：Its nucleosides Acid sequence is as shown in SEQ ID NO.13, H4-B3：Its nucleotide sequence is as shown in SEQ ID NO.14；And H4-FIP：Its Nucleotide sequence is as shown in SEQ ID NO.15, H4-BIP：Its nucleotide sequence is as shown in SEQ ID NO.16；

For the primer sets of IAV H5 areas fragment amplification, the primer sets include two pairs of primers, are respectively：H5-F3：Its nucleosides Acid sequence is as shown in SEQ ID NO.17, H5-B3：Its nucleotide sequence is as shown in SEQ ID NO.18；And H5-FIP：Its Nucleotide sequence is as shown in SEQ ID NO.19, H5-BIP：Its nucleotide sequence is as shown in SEQ ID NO.20；

For the primer sets of IAV H6 areas fragment amplification, the primer sets include two pairs of primers, are respectively：H6-F3：Its nucleosides Acid sequence is as shown in SEQ ID NO.21, H6-B3：Its nucleotide sequence is as shown in SEQ ID NO.22；And H6-FIP：Its Nucleotide sequence is as shown in SEQ ID NO.23, H6-BIP：Its nucleotide sequence is as shown in SEQ ID NO.24；

For the primer sets of IAV H7 areas fragment amplification, the primer sets include two pairs of primers, are respectively：H7-F3：Its nucleosides Acid sequence is as shown in SEQ ID NO.25, H7-B3：Its nucleotide sequence is as shown in SEQ ID NO.26；And H7-FIP：Its Nucleotide sequence is as shown in SEQ ID NO.27, H7-BIP：Its nucleotide sequence is as shown in SEQ ID NO.28；

For the primer sets of IAV H8 areas fragment amplification, the primer sets include two pairs of primers, are respectively：H8-F3：Its nucleosides Acid sequence is as shown in SEQ ID NO.29, H8-B3：Its nucleotide sequence is as shown in SEQ ID NO.30；And H8-FIP：Its Nucleotide sequence is as shown in SEQ ID NO.31, H8-BIP：Its nucleotide sequence is as shown in SEQ ID NO.32；

For the primer sets of IAV H9 areas fragment amplification, the primer sets include two pairs of primers, are respectively：H9-F3：Its nucleosides Acid sequence is as shown in SEQ ID NO.33, H9-B3：Its nucleotide sequence is as shown in SEQ ID NO.34；And H9-FIP：Its Nucleotide sequence is as shown in SEQ ID NO.35, H9-BIP：Its nucleotide sequence is as shown in SEQ ID NO.36；

For the primer sets of IAV H10 areas fragment amplification, the primer sets include two pairs of primers, are respectively：H10-F3：Its core Nucleotide sequence is as shown in SEQ ID NO.37, H10-B3：Its nucleotide sequence is as shown in SEQ ID NO.38；And H10- FIP：Its nucleotide sequence is as shown in SEQ ID NO.39, H10-BIP：Its nucleotide sequence is as shown in SEQ ID NO.40；

For the primer sets of IAV H11 areas fragment amplification, the primer sets include two pairs of primers, are respectively：H11-F3：Its core Nucleotide sequence is as shown in SEQ ID NO.41, H11-B3：Its nucleotide sequence is as shown in SEQ ID NO.42；And H11- FIP：Its nucleotide sequence is as shown in SEQ ID NO.43, H11-BIP：Its nucleotide sequence is as shown in SEQ ID NO.44；

For the primer sets of IAV H12 areas fragment amplification, the primer sets include two pairs of primers, are respectively：H12-F3：Its core Nucleotide sequence is as shown in SEQ ID NO.45, H12-B3：Its nucleotide sequence is as shown in SEQ ID NO.46；And H12- FIP：Its nucleotide sequence is as shown in SEQ ID NO.47, H12-BIP：Its nucleotide sequence is as shown in SEQ ID NO.48；

For the primer sets of IAV H13 areas fragment amplification, the primer sets include two pairs of primers, are respectively：H13-F3：Its core Nucleotide sequence is as shown in SEQ ID NO.49, H13-B3：Its nucleotide sequence is as shown in SEQ ID NO.50；And H13- FIP：Its nucleotide sequence is as shown in SEQ ID NO.51, H13-BIP：Its nucleotide sequence is as shown in SEQ ID NO.52；

For the primer sets of IAV H14 areas fragment amplification, the primer sets include two pairs of primers, are respectively：H14-F3：Its core Nucleotide sequence is as shown in SEQ ID NO.53, H14-B3：Its nucleotide sequence is as shown in SEQ ID NO.54；And H14- FIP：Its nucleotide sequence is as shown in SEQ ID NO.55, H14-BIP：Its nucleotide sequence is as shown in SEQ ID NO.56；

For the primer sets of IAV H15 areas fragment amplification, the primer sets include two pairs of primers, are respectively：H15-F3：Its core Nucleotide sequence is as shown in SEQ ID NO.57, H15-B3：Its nucleotide sequence is as shown in SEQ ID NO.58；And H15- FIP：Its nucleotide sequence is as shown in SEQ ID NO.59, H15-BIP：Its nucleotide sequence is as shown in SEQ ID NO.60；

For the primer sets of IAV H16 areas fragment amplification, the primer sets include two pairs of primers, are respectively：H16-F3：Its core Nucleotide sequence is as shown in SEQ ID NO.61, H16-B3：Its nucleotide sequence is as shown in SEQ ID NO.62；And H16- FIP：Its nucleotide sequence is as shown in SEQ ID NO.63, H16-BIP：Its nucleotide sequence is as shown in SEQ ID NO.64；

For the primer sets of IAV H17 areas fragment amplification, the primer sets include two pairs of primers, are respectively：H17-F3：Its core Nucleotide sequence is as shown in SEQ ID NO.65, H17-B3：Its nucleotide sequence is as shown in SEQ ID NO.66；And H17- FIP：Its nucleotide sequence is as shown in SEQ ID NO.67, H17-BIP：Its nucleotide sequence is as shown in SEQ ID NO.68；

For the primer sets of IAV H18 areas fragment amplification, the primer sets include two pairs of primers, are respectively：H18-F3：Its core Nucleotide sequence is as shown in SEQ ID NO.69, H18-B3：Its nucleotide sequence is as shown in SEQ ID NO.70；And H18- FIP：Its nucleotide sequence is as shown in SEQ ID NO.71, H18-BIP：Its nucleotide sequence is as shown in SEQ ID NO.72；

For the primer sets of IAV N1 areas fragment amplification, the primer sets include two pairs of primers, are respectively：N1-F3：Its nucleosides Sour nucleotide sequence is as shown in SEQ ID NO.73, N1-B3：Its nucleotide sequence is as shown in SEQ ID NO.74；And N1- FIP：Its nucleotide sequence is as shown in SEQ ID NO.75, N1-BIP：Its nucleotide sequence is as shown in SEQ ID NO.76；

For the primer sets of IAV N2 areas fragment amplification, the primer sets include two pairs of primers, are respectively：N2-F3：Its nucleosides Acid sequence is as shown in SEQ ID NO.77, N2-B3：Its nucleotide sequence is as shown in SEQ ID NO.78；And N2-FIP：Its Nucleotide sequence is as shown in SEQ ID NO.79, N2-BIP：Its nucleotide sequence is as shown in SEQ ID NO.80；

For the primer sets of IAV N3 areas fragment amplification, the primer sets include two pairs of primers, are respectively：N3-F3：Its nucleosides Acid sequence is as shown in SEQ ID NO.81, N3-B3：Its nucleotide sequence is as shown in SEQ ID NO.82；And N3-FIP：Its Nucleotide sequence is as shown in SEQ ID NO.83, N3-BIP：Its nucleotide sequence is as shown in SEQ ID NO.84；

For the primer sets of IAV N4 areas fragment amplification, the primer sets include two pairs of primers, are respectively：N4-F3：Its nucleosides Acid sequence is as shown in SEQ ID NO.85, N4-B3：Its nucleotide sequence is as shown in SEQ ID NO.86；And N4-FIP：Its Nucleotide sequence is as shown in SEQ ID NO.87, N4-BIP：Its nucleotide sequence is as shown in SEQ ID NO.88；

For the primer sets of IAV N5 areas fragment amplification, the primer sets include two pairs of primers, are respectively：N5-F3：Its nucleosides Acid sequence is as shown in SEQ ID NO.89, N5-B3：Its nucleotide sequence is as shown in SEQ ID NO.90；And N5-FIP：Its Nucleotide sequence is as shown in SEQ ID NO.91, N5-BIP：Its nucleotide sequence is as shown in SEQ ID NO.92；

For the primer sets of IAV N6 areas fragment amplification, the primer sets include two pairs of primers, are respectively：N6-F3：Its nucleosides Acid sequence is as shown in SEQ ID NO.93, N6-B3：Its nucleotide sequence is as shown in SEQ ID NO.94；And N6-FIP：Its Nucleotide sequence is as shown in SEQ ID NO.95, N6-BIP：Its nucleotide sequence is as shown in SEQ ID NO.96；

For the primer sets of IAV N7 areas fragment amplification, the primer sets include two pairs of primers, are respectively：N7-F3：Its nucleosides Acid sequence is as shown in SEQ ID NO.97, N7-B3：Its nucleotide sequence is as shown in SEQ ID NO.98；And N7-FIP：Its Nucleotide sequence is as shown in SEQ ID NO.99, N7-BIP：Its nucleotide sequence is as shown in SEQ ID NO.100；

For the primer sets of IAV N8 areas fragment amplification, the primer sets include two pairs of primers, are respectively：N8-F3：Its nucleosides Acid sequence is as shown in SEQ ID NO.101, N8-B3：Its nucleotide sequence is as shown in SEQ ID NO.102；And N8-FIP： Its nucleotide sequence is as shown in SEQ ID NO.103, N8-BIP：Its nucleotide sequence is as shown in SEQ ID NO.104；

For the primer sets of IAV N9 areas fragment amplification, the primer sets include two pairs of primers, are respectively：N9-F3：Its nucleosides Acid sequence is as shown in SEQ ID NO.105, N9-B3：Its nucleotide sequence is as shown in SEQ ID NO.106；And N9-FIP： Its nucleotide sequence is as shown in SEQ ID NO.107, N9-BIP：Its nucleotide sequence is as shown in SEQ ID NO.108；

For the primer sets of IAV N10 areas fragment amplification, the primer sets include two pairs of primers, are respectively：N10-F3：Its core Nucleotide sequence is as shown in SEQ ID NO.109, N10-B3：Its nucleotide sequence is as shown in SEQ ID NO.110；And N10- FIP：Its nucleotide sequence is as shown in SEQ ID NO.111, N10-BIP：Its nucleotide sequence is as shown in SEQ ID NO.112；

For the primer sets of IAV N11 areas fragment amplification, the primer sets include two pairs of primers, are respectively：N11-F3：Its core Nucleotide sequence is as shown in SEQ ID NO.113, N11-B3：Its nucleotide sequence is as shown in SEQ ID NO.114；And N11- FIP：Its nucleotide sequence is as shown in SEQ ID NO.115, N11-BIP：Its nucleotide sequence is as shown in SEQ ID NO.116.

2. LAMP primer group according to claim 1, it is characterised in that the primer sets include H1~H18 for IAV One group in the primer sets of area's fragment amplification or several groups, and, for one in the primer sets of IAV N1~N11 areas fragment amplification Kind is several.

3. a kind of kit for being used to detect influenza A virus, it is characterised in that the kit includes claim 1 or 2 Described LAMP primer group.

4. kit according to claim 3, it is characterised in that the kit also includes LAMP reaction solutions and LAMP shows Color liquid.

5. kit according to claim 4, it is characterised in that the LAMP reaction solutions include：Bst polymerases, AMV are anti- Transcriptase, LAMP reaction buffers and ultra-pure water.

6. a kind of influenza A virus detection method of non-disease diagnostic purpose, comprises the following steps：

1) take working solution and LAMP reaction solutions, ultra-pure water, the LAMP of sample of nucleic acid, claim 1 or 2 the LAMP primer group close Sealing liquid mixes, and amplification reaction solution is made；

2) obtained amplification reaction solution is taken, 60~65 DEG C of 20~80min of reaction, whether is contained according to colour developing result judgement sample IAV viruses.

7. detection method according to claim 6, it is characterised in that in the working solution of the LAMP primer group, primer H1- F3 and H1-B3 concentration is 0.2~0.4 μM, and primer H1-FIP and H1-BIP concentration are 1~2 μM；

The concentration of the primer H7-F3 and H7-B3 is 0.2~0.4 μM, and primer H7FIP and H7-BIP concentration are 1~2 μ M；

The concentration of the primer H10-F3 and H10-B3 is 0.2~0.4 μM, and primer H10-FIP and H10-BIP concentration are 1~2 μM；

The concentration of the primer H11-F3 and H11-B3 is 0.2~0.4 μM, and primer H11-FIP and H11-BIP concentration are 1~2 μM；

The concentration of the primer H12-F3 and H12-B3 is 0.2~0.4 μM, and primer H12-FIP and H12-BIP concentration are 1~2 μM；

The concentration of the primer H13-F3 and H13-B3 is 0.2~0.4 μM, and primer H13-FIP and H13-BIP concentration are 1~2 μM；

The concentration of the primer H14-F3 and H14-B3 is 0.2~0.4 μM, and primer H14-FIP and H14-BIP concentration are 1~2 μM；

The concentration of the primer H15-F3 and H15-B3 is 0.2~0.4 μM, and primer H15-FIP and H15-BIP concentration are 1~2 μM；

The concentration of the primer H16-F3 and H16-B3 is 0.2~0.4 μM, and primer H16-FIP and H16-BIP concentration are 1~2 μM；

The concentration of the primer H17-F3 and H17-B3 is 0.2~0.4 μM, and primer H17-FIP and H17-BIP concentration are 1~2 μM；

The concentration of the primer H18-F3 and H18-B3 is 0.2~0.4 μM, and primer H18-FIP and H18-BIP concentration are 1~2 μM；

The concentration of the primer N7-F3 and N7-B3 is 0.2~0.4 μM, and primer N7FIP and N7-BIP concentration are 1~2 μ M；

The concentration of the primer N10-F3 and N10-B3 is 0.2~0.4 μM, and primer N10-FIP and N10-BIP concentration are 1~2 μM；

The concentration of the primer N11-F3 and N11-B3 is 0.2~0.4 μM, and primer N11-FIP and N11-BIP concentration are 1~2 μM.

8. application of the LAMP primer group of claim 1 or 2 in the IAV Viral diagnosis of non-disease diagnostic purpose.

9. application of any kits of claim 3-5 in the IAV Viral diagnosis of non-disease diagnostic purpose.