CN1906301A - HIV-1 Indian subtype C vaccine constructs for use in humans - Google Patents
HIV-1 Indian subtype C vaccine constructs for use in humans Download PDFInfo
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- CN1906301A CN1906301A CNA2004800404194A CN200480040419A CN1906301A CN 1906301 A CN1906301 A CN 1906301A CN A2004800404194 A CNA2004800404194 A CN A2004800404194A CN 200480040419 A CN200480040419 A CN 200480040419A CN 1906301 A CN1906301 A CN 1906301A
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
- A61K2039/525—Virus
- A61K2039/5256—Virus expressing foreign proteins
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
- A61K2039/53—DNA (RNA) vaccination
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- C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
- C12N2710/00011—Details
- C12N2710/24011—Poxviridae
- C12N2710/24111—Orthopoxvirus, e.g. vaccinia virus, variola
- C12N2710/24141—Use of virus, viral particle or viral elements as a vector
- C12N2710/24143—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N2740/00—Reverse transcribing RNA viruses
- C12N2740/00011—Details
- C12N2740/10011—Retroviridae
- C12N2740/16011—Human Immunodeficiency Virus, HIV
- C12N2740/16111—Human Immunodeficiency Virus, HIV concerning HIV env
- C12N2740/16122—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2740/00—Reverse transcribing RNA viruses
- C12N2740/00011—Details
- C12N2740/10011—Retroviridae
- C12N2740/16011—Human Immunodeficiency Virus, HIV
- C12N2740/16311—Human Immunodeficiency Virus, HIV concerning HIV regulatory proteins
- C12N2740/16322—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
Abstract
Human Immunodeficiency virus (HIV) vaccine constructs to combat Indian subtype C HIV-1 Infection In humans comprising structural and non structural genes of Indian strain of HIV-1 subtype C mounted onto a DNA plasmid vector.
Description
Invention field
The present invention relates to development of HIV-1 vaccine candidate object and preparation method thereof.Particularly, the present invention relates to India HIV-1C hypotype construct, it comprises coating (gp120) and capsid (gag-proteolytic enzyme), the nef that clones and the tat gene and the virus vector of HIV-1C hypotype in the plasmid DNA carrier.
Background of invention
AIDS threatens one of the most serious human transmissible disease now.Therefore, a kind of vaccine safely and effectively of research and development obviously is to stop the task of top priority of AIDS popular.
All organisms, especially human, have immunity system, its protection body resistance comprises the invasion of the various microorganisms of virus.Yet HIV can invade immune cell and destroy immunity system.Thereby virus is duplicated under the situation that does not have monitoring in vivo.
The vaccine of a kind of effective opposing HIV can stop the propagation of virus in the mankind, and this has become a common objective of scientific circles.Effectively vaccine should be induced long-term and intensive immunne response, can resist HIV so in advance and invade human body.
The method that is used for virus vaccines in the past is as the live virus construct of use attenuation or the virus of deactivation, succeed as poliovirus or Measles virus although these are used for other virus infection, for HIV, then be faced with an outstanding safety problem.
The another kind of method that has been used to prepare the HIV vaccine is a recombinant DNA technology.This method allows scientist to use the part of HIV gene to come induce immune response.These genes are arranged on the vector dna molecule that is called carrier, are injected into acceptor immune response interior and reactance HIV in its body then; The security of height so just is provided.Similarly, these genes also can be arranged on the virus vector, just can induce the HIV specific immune response in animal body at that time when injecting.
Goal of the invention
An object of the present invention is to provide a kind of vaccine that is used for anti-human immunodeficiency virus 1 type (HIV-1) the India C hypotype of human body.
Another object of the present invention provides a kind of vaccine that is used for effectively resisting HIV-1 India C subtype viral infection.
Other purpose and advantage of the present invention will further describe in the following description.
Specification sheets begin the place and hereinafter, will be clear that specification sheets subsequently only is to have illustrated the application's specific form.But above-mentioned specific form of the present invention only is an exemplary embodiment, and not should be understood to be the exemplary embodiment that the present invention instructs, and should not think restrictive.
Summary of the invention
According to the present invention, here provide a kind of and be used to resist the HIV vaccine constructs that the HIV-1C hypotype infects based on recombinant DNA technology, it comprises the structure gene (coating and gap-proteolytic enzyme) and the nonstructural gene (nef and tat) of the HIV-1C hypotype India strain that the mankind are codon optimized, and it is set on plasmid DNA carrier and the virus vector.
The structure gene of described in the present invention India HIV-1C hypotype, gp120 and gap-proteolytic enzyme are the parts of HIV-1C subtype virus nucleic acid.Nonstructural gene is that virus replication is necessary.The sequence of these genes is modified according to the occurrence rate of human codon, but does not change its aminoacid sequence.
After codon optimized structure gene is synthetic, be set on plasmid DNA carrier or the virus vector, it carries these genes and enters in the human body cell.Processing produces the albumen that does not influence cell function to these genes through cell then.
According to the present invention, here the coating gp120, gag-proteolytic enzyme, the nef of India HIV-1C hypotype and the method for gene constructed plasmid DNA vaccine of tat and recombinant viral vector vaccine constructs that also provide a kind of son that accesses to your password to optimize, it can cause the HIV-1 specific immune response in these animals after injecting these vaccine constructs for mouse and monkey.
Detailed Description Of The Invention
1. be present in the amplification of coating gp120 gene, gag-proteinase gene, nef gene and the tat gene of the HIV-1C hypotype (India's strain) in the peripheral blood of three infected asymptomatic individualities.
Use polymerase chain reaction (PCR) technology and use suitable primer amplification separation from the gp120 gene region of the 1.4kb of peripheral blood lymphocytes (PBMCs) DNA of the asymptomatic individuality of three HIV infection and the gag-proteinase gene zone of 1.8kb.Briefly, PBMCs obtains from the peripheral blood separation by ficoll-hypaque density gradient centrifugation.Use ethanol sedimentation DNA then by handling dissolving PBMCs with Proteinase K, from these cells, extract DNA at 72 ℃.Using spin this DNA of column purification then and being dissolved in pH is in 8.0 the TE damping fluid.
For amplification gene, will be by damping fluid, forward and reverse primer, dNTPs (dATPs, dTTPs, dCTPs, dGTPs), MgCl
2The reaction mixture of forming with the Taq archaeal dna polymerase adds in the template DNA (extracting from PBMCs) of thermally denature.The cycling condition of PCR is as follows: 92 ℃ of following sex change 20 seconds, 52 ℃ of following annealing 40 seconds, 68 ℃ of following extensions 4 minutes.Circulate in the manner described above 25 times, then 72 ℃ of last extensions 15 minutes.With the DNA of ethanol sedimentation amplification, then by the gel electrophoresis purifying.The DNA prepared product of these amplifications contains 2 coating gp120 genes and 1 gag-proteinase gene, 1 nef gene and 1 tat gene.
2. order-checking
Use the primer walking method to instruct sequencing reaction to come these 5 amplification genes are checked order.The DNA of these amplification genes comes purifying by ethanol sedimentation, uses Big Dye to stop test kit to each DNA prepared product and carries out sequencing reaction and use PCR System 2400 to carry out cycle sequencing.Then with the sequence data classification, comparison and the analysis that obtain.According to human codon occurrence rate manually carry out codon optimization thereafter.
3. codon optimization
According to human codon occurrence rate these 5 genes are carried out the codon optimization in the hope of better being expressed and causing stable immunne response in human body.When carrying out the codon optimization, should carefully not upset corresponding amino acid code.Under the situation that does not upset described aminoacid sequence, remove " INS " sequences all in the gag-proteinase gene, keep the optimization (stem-ring-stem) that sequence between the nt 1276 to nt 1477 is not carried out codon simultaneously.
4. the structure of codon optimization gene
Make up described codon optimization gene by the pcr amplification method.Gene to this structure checks order on automatic sequencer then.The sequence data of this synthetic gene and expected sequence relatively, these two sequences have 100% homology.With these codon optimization unnamed genes be: env-gp120-29692CO, env-gp 120-49426CO, gap-proteolytic enzyme 49587CO, IND-tatCO, IND-nefCO.
5. the clone of codon optimization gene in plasmid vector
After described 5 genes are synthetic, it is cloned in the mammalian expression vector.Test the plasmid that generates, NK-29692CO, NK4426CO and NK-49587CO, NK49587CO, NK-IND-tatCO and the NK-IND-nefCO vivoexpression in the 293T cell of people source then.With these plasmid transfections 293T cell.After 72 hours, detect expression of gene described in these transfectional cells by immunoblotting.In the lysate of cell and culture supernatant, all detect the particular bands of antigen expressed.Can see the film that virus-like particle and cytolemma one deck on every side are made up of gp120 antigen by transmission electron microscope, further confirm described codon optimization expression of gene.
6. the clone of codon optimization gene in virus vector
After described 5 genes are synthetic, it is cloned among the plasmid shuttle vectors pSC65.The recombinant plasmid that generates is used to make up the recombinant MVA that carries HIV-1 India C hypotype structure gene: VV-29692CO, VV49426CO, VV-49587CO, VV-IND-tatCO and VV-IDN-nefCO.Detect the vivoexpression of these recombinant chous in cell culture system (BHK-21) by western blot analysis, p-24 antigen measuring, indirect immunofluorescence assay (IFA) and transmission electron microscopy (TEM).Western blotting has shown that corresponding HIV specific proteins, IFA have shown the apple green fluorescence on the infected B HK-21 cell surface and TEM has shown the blastogenesis from the virus-like particle of cells infected.
7. for the immunogenicity research of monkey
Use and add the next immune Macaca radiata (bonnet monkey) of strong method first.HIV plasmid DNA vaccine constructs is used for construction of recombinant plasmid body NK-29692CO, NK-49426CO, NK-49587CO, NK-IND-tatCO and the NK-IND-nefCO of single dose animal being carried out intradermal immunization first.All around, intracutaneous gives recombinant MVA construct VV-29692CO, VV-49426CO, VV-49587CO, VV-IND-tatCO and the VV-IND-nefCO of single booster dose.Then, in these animals, measure immunne response at the proteic HIV-1C hypospecificity of HIV-1.Detect dual mode, antibody and the cell response of immunne response respectively by ELISA and ELISPOT.This animal by immunity has produced strong HIV-1C subtype specificity antibody and cell-mediated immune responses, even still has embodiment after immune 42 weeks.
These data show that these above-mentioned vaccine candidate objects have as the immune potentiality that are used for the HIV-1C subgroup vaccine material standed for of human body.
7. experimental program
A) pcr amplification is obtained from coating gp120, gap-proteolytic enzyme, tat and the nef gene of HIV-1C hypotype (India's strain)
Extract DNA from the PBMCs of HIV-1 infected individuals
↓
The complete coating gp120 of pcr amplification is with the gagpol gene and separate amplified production
↓
TA is cloned among the pGEMTeasy with amplified production
↓
Screen recombinant chou through the following steps
※ carries out PCR to env gp120, gag and proteolytic enzyme, tat and nef group
The ※ restrictive diges-tion
B) to env gp120 and the order-checking of gag proteinase gene
With the plasmid DNA of purifying as the template DNA that is used for sequencing reaction
↓
Use different forwards and reverse primer to carry out sequencing reaction
↓
Using Big Dye to stop test kit checks order
↓
The continuous fragment of 500-600bp is spliced into gene
↓
Operation Blast measures homology
↓
Adopting maximum comparability method and contiguous method of attachment to carry out system analyzes
↓
Determine structural motif and read frame
C) structure of codon optimization and optimization gene
D) the optimized gene of clone's codon in the plasmid DNA carrier
I) in the gap-proteolytic enzyme-49597CO gene clone plasmid dna vector with HIV-1C hypotype (India's strain).
To carry out pcr amplification from pGEM-T49587CO clone's gap proteinase gene
↓
PCR product flush end is cloned among the plasmid DNA pNK14
↓
By following steps screening recombinant chou
※ carries out PCR to gag and proteinase gene
The ※ restrictive diges-tion
↓
Select NK14-49587CO
II) with the gp120 29692CO gene clone of HIV-1C hypotype (India's strain) in plasmid DNA carrier pNK14.
To carry out pcr amplification from pGEM-T-29692CO clone's env gp120 296920CO gene
↓
Flush end is cloned among the plasmid DNA carrier pNK14
↓
By following steps screening recombinant chou
※ carries out PCR to coating gp.120 gene
The ※ restrictive diges-tion
↓
Select NK14-29692CO
III) plasmid DNA carrier pNK14 is arrived in the env gp120 49426CO gene clone of HIV-1C hypotype (India's strain).
To carry out pcr amplification from pGEM-T-49426CO clone's env gp120 49426CO gene
↓
Flush end is cloned among the plasmid DNA carrier pNK14
↓
By following steps screening recombinant chou
※ carries out PCR to coating gp.120 gene
The ※ restrictive diges-tion
↓
Select NK14-49426CO
IV) pSC gag proteolytic enzyme 49587, env gp120 29692 and env gp120 49426 genes of being cloned in the plasmid DNA carrier are checked order.
The plasmid DNA of purifying is as the template DNA that is used for sequencing reaction
↓
Use different forwards and reverse primer to carry out sequencing reaction
↓
Using Big Dye to stop test kit checks order
↓
The continuous fragment of 400-500bp is spliced into gene
E) construction of recombinant virus vector construction body
I) shuttle vectors is arrived in the gap-proteolytic enzyme-49597CO gene clone of HIV-1C hypotype (India's strain), among the pSC65.
To carry out pcr amplification from pGEM-T-49587CO clone's gap proteinase gene
↓
PCR product flush end is cloned into shuttle vectors, among the pSC65
↓
By following steps screening recombinant chou
※ carries out PCR to gag and proteinase gene
The ※ restrictive diges-tion
↓
Select pSCgp proteolytic enzyme 49587
II) shuttle vectors is arrived in the gp120 29692CO gene clone of HIV-1C hypotype (India's strain), among the pSC65.
To carry out pcr amplification from pGEM-T-29692CO clone's env gap120 296920 genes
↓
Flush end is cloned among the shuttle vectors pSC65
↓
By following steps screening recombinant chou
※ carries out PCR to coating gP.120 gene
The ※ restrictive diges-tion
↓
Select pSC29692CO
III) shuttle vectors is arrived in the env gp120 49426CO gene clone of HIV-1C hypotype (India's strain), among the pSC65.
To carry out pcr amplification from pGEM-T-49426CO clone's env gp120 49426CO gene
↓
Flush end is cloned among the shuttle vectors pSC 65
↓
By following steps screening recombinant chou
※ carries out PCR to coating gp.120 gene
The ※ restrictive diges-tion
↓
Select pSC49426CO
IV) pSC gag proteolytic enzyme 49587, env gp120 29692 and env gp120 49426 genes of being cloned among the shuttle vectors pSC65 are checked order.
The plasmid DNA of purifying is as the template DNA that is used for sequencing reaction
↓
Use different forwards and reverse primer to carry out sequencing reaction
↓
Using Big Dye to stop test kit checks order
↓
The continuous fragment of 400-500bp is spliced into gene
V) express the generation of the recombinant MVA of gag proteolytic enzyme and env gp120 gene respectively
VI) purifying of recombinant MVA
The 6 blue plaque purifications of taking turns
↓
The recombinant virus of amplification purification
↓
Granulation and and purified virus on 36% sucrose layer
F) in the Bonnet monkey, study immunogenicity
Claims
(according to the modification of the 19th of treaty)
1. be used for resisting human immunodeficiency virus (HIV) vaccine constructs of India C hypotype HIV-1 in the infection of human body, it contains the structure gene and the nonstructural gene of India's strain of the HIV-1C hypotype that is arranged on the DNA plasmid vector.
2. be used for resisting human immunodeficiency virus (HIV) vaccine constructs of India C hypotype HIV-1 in the infection of human body, it contains the structure gene and the nonstructural gene of India's strain of the HIV-1C hypotype that is arranged on the virus vector.
3. according to claim 1 and 2 described HIV-1 vaccine constructs, wherein said structure gene is coating gp120 gene and gag-proteinase gene.
4. according to claim 1 and 2 described HIV-1 vaccine constructs, wherein said nonstructural gene is tat and nef gene.
5. according to claim 1 and 2 described HIV-1 vaccine constructs, the wherein said coating gp120 gene that is obtained from India's strain 29692 is env-gp120-29692.
6. according to claim 1 and 2 described HIV-1 vaccine constructs, the wherein said coating gp120 gene that is obtained from India's strain 49426 is env-gp120-49426.
7. according to claim 1 and 2 described HIV-1 vaccine constructs, the wherein said gag proteinase gene that is obtained from India's strain 49587 is a gag-proteolytic enzyme-49587.
8. according to claim 1 and 2 described HIV-1 vaccine constructs, wherein said tat gene is obtained from the consensus sequence of HIV-1 India C subtype isolates.
9. according to claim 1 and 2 described HIV-1 vaccine constructs, wherein said nef gene is the consensus sequence that is obtained from HIV-1 India C subtype isolates.
10. according to claim 1 and 2 described HIV-1 vaccine constructs, the env gene of wherein said HIV-1C hypotype is codon-optimized and obtain coating gp120-29692CO gene.
11. according to claim 1 and 2 described HIV-1 vaccine constructs, the env gene of wherein said HIV-1C hypotype is codon-optimized and obtain coating gp120 49426-CO gene.
12. according to claim 1 and 2 described HIV-1 vaccine constructs, the gag-proteinase gene of wherein said HIV-1C hypotype is codon-optimized and obtain gag-proteolytic enzyme 49587-CO gene.
13. according to claim 1 and 2 described HIV-1 vaccine constructs, the tat gene of wherein said HIV-1C hypotype is codon-optimized and obtain the IND-tatCO gene.
14. according to claim 1 and 2 described HIV-1 vaccine constructs, the nef gene of wherein said HIV-1C hypotype is codon-optimized and obtain the IND-nefCO gene.
15.HIV-1C the subgroup vaccine construct, it contains recombinant plasmid dna vaccine constructs as described herein basically and recombinant viral vector vaccine constructs.
Claims (24)
1. be used for resisting human immunodeficiency virus (HIV) vaccine constructs of India C hypotype HIV-1 in the infection of human body, it contains the structure gene and the nonstructural gene of India's strain of the HIV-1C hypotype that is arranged on the DNA plasmid vector.
2. be used for resisting human immunodeficiency virus (HIV) vaccine constructs of India C hypotype HIV-1 in the infection of human body, it contains the structure gene and the nonstructural gene of India's strain of the HIV-1C hypotype that is arranged on the virus vector.
3. according to claim 1 and 2 described HIV-1 vaccine constructs, wherein said structure gene is coating gp120 gene and gag-proteinase gene.
4. according to claim 1 and 2 described HIV-1 vaccine constructs, wherein said nonstructural gene is tat and nef gene.
5. according to claim 1 and 2 described HIV-1 vaccine constructs, the wherein said coating gp120 gene that is obtained from India's strain 29692 is env-gp-120-29692.
6. according to claim 1 and 2 described HIV-1 vaccine constructs, the wherein said coating gp120 gene that is obtained from India's strain 49426 is env-gp-120-49426.
7. according to claim 1 and 2 described HIV-1 vaccine constructs, the wherein said gag proteinase gene that is obtained from India's strain 49587 is a gag-proteolytic enzyme-49587.
8. according to claim 1 and 2 described HIV-1 vaccine constructs, wherein said tat gene is the consensus sequence that is obtained from HIV-1 India C subtype isolates.
9. according to claim 1 and 2 described HIV-1 vaccine constructs, wherein said nef gene is the consensus sequence that is obtained from HIV-1 India C subtype isolates.
10. according to claim 1 and 2 described HIV-1 vaccine constructs, the env gene of wherein said HIV-1C hypotype is codon-optimized and obtain coating gp120-29692CO gene.
11. according to claim 1 and 2 described HIV-1 vaccine constructs, the env gene codon optimization of wherein said HIV-1C hypotype and obtain coating gp120 49426-CO gene.
12. according to claim 1 and 2 described HIV-1 vaccine constructs, the gag-proteinase gene of wherein said HIV-1C hypotype is codon-optimized and obtain gag-proteolytic enzyme 49587-CO gene.
13. according to claim 1 and 2 described HIV-1 vaccine constructs, the tat gene of wherein said HIV-1C hypotype is codon-optimized and obtain the IND-tatCO gene.
14. according to claim 1 and 2 described HIV-1 vaccine constructs, the nef gene of wherein said HIV-1C hypotype is codon-optimized and obtain the IND-nefCO gene.
15. one kind is used the coating gp120 of India HIV-1C hypotype and the method that gag-proteolytic enzyme, tat and nef gene prepare the plasmid DNA vaccine constructs, it comprises:
HIV-1C hypotype env gp120 and gag-proteolytic enzyme, tat and the nef gene that will be present in the virus in the infected individuals peripheral blood cells carry out pcr amplification; To 2 env genes, 1 gag-proteinase gene, 1 tat gene and 1 nef gene sequencing; Synthetic codon optimized gene; The gene of described optimization is arranged on the plasmid DNA carrier; In the bonnet monkey, cause immunne response by injection recombinant plasmid dna vaccine constructs.
16. coating gp120, gag-proteolytic enzyme, tat and a nef gene that uses India HIV-1C hypotype prepares the method for recombinant viral vector vaccine constructs, it comprises:
To be present in the HIV-1C hypotype env gp120 of the virus in the infected individuals peripheral blood cells, gag-proteolytic enzyme, tat and nef gene carry out pcr amplification; To 2 env genes, 1 gag-proteinase gene, 1 tat gene and 1 nef gene sequencing; Synthetic codon optimized gene; The gene of described optimization is arranged at the plasmid shuttle vectors, on the pSC65; Use is connected with the shuttle vectors transfection MVA infected B HK-21 cell of HIV-1 codon optimization gene; Select substratum to select to contain the recombinant MVA of HIV-1 gene with BuDR and X-gal; Purification of Recombinant virus; In the bonnet monkey, cause immunne response by injection recombinant viral vector vaccine constructs.
17. according to claim 15 and 16 described methods, wherein said gene amplification step is the polymerase chain reaction.
18. according to claim 15 and 16 described methods, wherein said peripheral blood cells separating step be undertaken by ficoll-hypaque density gradient centrifugation and handle the described cell of dissolving by the proteolytic enzyme under 72 ℃-K and use solvent deposition DNA subsequently, from described cell, extract DNA.
19. according to claim 15 and 16 described methods, wherein said gene checks order by being cloned among the plasmid vector pGEMTeasy.
20. according to claim 15 and 16 described methods, wherein said gene has carried out the codon optimization according to human codon occurrence rate.
21. method according to claim 15, wherein said recombinant plasmid NK-29692CO, NK-49426CO, NK49587CO, NK-IND-tatCO and NK-IND-nefCO are used to study the immunogenicity of bonnet monkey.
22. method according to claim 15, wherein said recombinant MVA, promptly VV-29692CO, VV-49426CO, VV-49587CO, VV-IND-tatCO and VV-IND-nefCO are used to study the immunogenicity of bonnet monkey.
23.HIV-1C the subgroup vaccine construct, it contains recombinant plasmid dna vaccine constructs as described herein basically and recombinant viral vector vaccine constructs.
24. a method for preparing HIV-1C subgroup vaccine construct, described construct contain recombinant plasmid dna vaccine constructs as described herein basically and recombinant viral vector vaccine constructs.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| IN63/DEL/04 | 2004-01-16 | ||
| IN63DE2004 | 2004-01-16 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1906301A true CN1906301A (en) | 2007-01-31 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNA2004800404194A Pending CN1906301A (en) | 2004-01-16 | 2004-09-10 | HIV-1 Indian subtype C vaccine constructs for use in humans |
Country Status (3)
| Country | Link |
|---|---|
| CN (1) | CN1906301A (en) |
| WO (1) | WO2005068634A1 (en) |
| ZA (1) | ZA200605498B (en) |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| TW200613554A (en) | 2004-06-17 | 2006-05-01 | Wyeth Corp | Plasmid having three complete transcriptional units and immunogenic compositions for inducing an immune response to HIV |
-
2004
- 2004-09-10 WO PCT/IN2004/000284 patent/WO2005068634A1/en active Application Filing
- 2004-09-10 CN CNA2004800404194A patent/CN1906301A/en active Pending
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- 2006-07-04 ZA ZA200605498A patent/ZA200605498B/en unknown
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| Publication number | Publication date |
|---|---|
| WO2005068634A1 (en) | 2005-07-28 |
| ZA200605498B (en) | 2007-09-26 |
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