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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 problems such as inconsistency of genotypes of epidemic strains, and achieve the effect of high reproduction performance

Active Publication Date: 2012-11-14
YANGZHOU UNIV
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Problems solved by technology

[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

Method used

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  • Gene VIb subtype Rubulavirus Newcastle disease virus attenuated strain VIbI4 and construction method thereof
  • Gene VIb subtype Rubulavirus Newcastle disease virus attenuated strain VIbI4 and construction method thereof
  • Gene VIb subtype Rubulavirus Newcastle disease virus attenuated strain VIbI4 and construction method thereof

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Embodiment Construction

[0032] The gene VIb subtype pigeon-derived Newcastle disease virus attenuated strain VIbI4 constructed by the present invention was preserved in the General Microbiology Center of China Microbiological Culture Collection Management Committee on May 23, 2012. The address of the preservation unit is: Institute of Microbiology, Chinese Academy of Sciences, and its preservation number CGMCC No: 6149, its length: 15192bp.

[0033] Construction Step 1: Construction of Attenuated Full-length Expression Cloning of Pigeon Newcastle Disease Virus

[0034] Biomaterial 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, et al. Infection Construction of sexual molecular clones and virus rescue. Acta Virus. 2012, 28 (1), 67-72.] Constructed and preserved by the Laboratory of Livestock and Poultry Infectious Diseases, Ministry of Agriculture, Yangzh...

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Abstract

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 invention relates to the application of reverse genetic technology to produce a gene VIb subtype Newcastle disease virus attenuated strain VIbI4 for making vaccines. Background technique [0002] Reverse genetics manipulation technique (Reverse genetics manipulation technique) refers to the construction of infectious molecular clones of RNA viruses in vitro, the reverse transcription of viral genomic RNA into cDNA, and various in vitro artificial manipulations on the DNA molecular level. Genomic cDNA and various auxiliary proteins to assemble a new RNA virus [Leyrer S, Neubert WJ, SedlmeierR. Rapid and efficient recovery of Sendai virus from cDNA: factors influencing recombinant virus rescue. J Virol Methods 1998, 75( 1):47-58.], also known as full-length infectious cDNA cloning technology, and often referred to as "virus rescue". Since the "rescued" RNA virus is derived from cDNA clones, various in vitro artificial operations can be performed on ...

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Application Information

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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
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