Gene VIb subtype Rubulavirus Newcastle disease virus attenuated strain VIbI4 and construction method thereof
A technology of Newcastle disease virus and construction method, which is applied in the field of gene VIb subtype Newcastle disease virus attenuated strain VIbI4 and its construction, can solve the problems of inconsistency of genotypes of epidemic strains and the like, and achieve the effect of high reproduction performance
Active Publication Date: 2013-11-06
YANGZHOU UNIV
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[0008] The first purpose of the present invention is to invent a gene VIb subtype pigeon-derived Newcastle disease virus attenuated strain, thereby solving the problem of inconsistency between the currently used vaccine strain and the pigeon Newcastle disease epidemic strain genotype
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[0032] The gene Ⅵb subtype pigeon-derived Newcastle disease virus attenuated strain ⅥbI4 constructed in the present invention was deposited in the General Microbiology Center of the Chinese Microbial Culture Collection Management Committee on May 23, 2012. The address of the depository: Institute of Microbiology, Chinese Academy of Sciences, its deposit number CGMCC No: 6149, its length is: 15192bp.
[0033] Construction Step 1: Construction of a full-length expression clone attenuated by pigeon Newcastle disease virus
[0034] Biological material preparation:
[0035] The full-length expression clone pTVT / 071204 of Newcastle disease virus JS / 07 / 04 / Pi strain, and its intermediate plasmids P34 and P58 [Zhu Yanmei, Hu Zenglei, Song Qingqing, etc. Pigeon source Newcastle disease virus JS / 07 / 04 / Pi strain infection Construction of Sexual Molecular Clones and Virus Rescue. Journal of Virology. 2012, 28(1), 67-72.] Constructed and preserved by the Animal Infectious Disease Laboratory of th...
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The invention relates to gene VIb subtype Avian pneumo-encephalitis virus attenuated strain VIbI4 and a construction method thereof. The preserving number of the gene VIb subtype scriber set Avian pneumo-encephalitis virus attenuated strain VIbI4 is CGMCCNo: 6149. The gene VIb subtype Rubulavirus Newcastle disease virus attenuated strain VIbI4 and the construction method thereof relate to a technology for applying reverse genetics; and the method comprises the following steps: carrying out mutation on an F gene by using a transcription carrier Ptvt / 071204 containing Avian pneumo-encephalitis virus JS / 07 / 04 / Pi full gene genome from pigeons so as to obtain to a transcription carrier ApTVT / 071204, and then substituting corresponding parts on the ApTVT / 071204 by the NP, P and L genetic fragments of Avian pneumo-encephalitis virus rNDV / I4 so as to obtain recombinant plasmids ApTVT / VIbI4, wherein the plasmids successfully save a recombinant virus VIbI4 after transfecting a BSR-T7 / 5 cell together with auxiliary plasmids. The virus is relatively high in propagating titre, is suitable for large-scale production of vaccines and can be used for manufacturing vaccines.
Description
Technical field [0001] The present invention relates to the application of reverse genetic technology to produce an attenuated strain VIbI4 of gene VIb subtype Newcastle disease virus, which is used to manufacture vaccines. Background technique [0002] Reverse genetics manipulation technique refers to the construction of infectious molecular clones of RNA viruses in vitro, reverse transcription of viral genome RNA into cDNA, and various in vitro manual manipulations on the DNA molecular level. Genome cDNA and various accessory proteins to assemble a new RNA virus [Leyrer S, Neubert WJ, Sedlmeier R. Rapid and efficient recovery of Sendai virus from cDNA: factors influencing recombinant virus rescue. J Virol Methods 1998, 75( 1):47-58.], also called full-length infectious cDNA cloning technology, and often called "virus rescue". Because the RNA virus "rescued" ultimately comes from cDNA clones, various in vitro manual operations can be performed on the RNA virus genome at the DNA...
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IPC IPC(8): C12N7/01C12N15/85C12R1/93
Inventor 刘秀梵朱艳梅胡增垒胡顺林王晓泉
Owner YANGZHOU UNIV