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Gene VII type new castle disease virus weakening strain A-NDV-VII and construction method thereof

A technology of A-NDV-VII, Newcastle disease virus, applied in the direction of virus/phage, biochemical equipment and method, inactivation/attenuation, etc., can solve the problem of inconsistency of genotype, achieve high reproductive titer, wide application Foreground effect

Active Publication Date: 2010-09-08
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The first object of the present invention is to invent a weakened strain of gene VII type Newcastle disease virus, thereby solving the problem that the genotypes of currently used vaccine strains are inconsistent with epidemic strains

Method used

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  • Gene VII type new castle disease virus weakening strain A-NDV-VII and construction method thereof
  • Gene VII type new castle disease virus weakening strain A-NDV-VII and construction method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0060] Construction Step 1: Genome Modification of ZJ1 Strain

[0061] Biomaterial preparation:

[0062] Newcastle disease virus ZJ1 strain was isolated and preserved by the Key Open Laboratory of Livestock and Poultry Infectious Diseases, Ministry of Agriculture, Yangzhou University.

[0063] pNDV / ZJ1: Constructed by the Laboratory of Livestock and Poultry Infectious Diseases of the Ministry of Agriculture of Yangzhou University, and sold publicly.

[0064] pCR2.1 vector: purchased from Invitrogen.

[0065] Three pairs of primers were designed to amplify a fragment of 2849-9552nt about 6700bp in the genome of ZJ1 strain. The primer sequences are as follows:

[0066] Primer number (position of amplified fragment)

Primer sequence (endonuclease)

R1 (2849-5223nt) R2 (5218-7012nt) R3 (7008-9552nt)

5'-GGGTGAAATGACGCTCAATAAACTCTC-3' 5'-ATGGTCTCATCTGTGGCCCGAATACT-3'(Bsa I) 5'-ATACGTCTCACAGATCACCTCCCCTGCATTAAC-3'(Bsm B I) 5'-GTC4GATCTCTGCAACCGGATAGTATCACA-3'(...

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Abstract

A VII gene type of an attenuated strain of Newcastle disease virus A-NDV-VII and a construction method are disclosed. The invention relates to the application of reverse genetics technique. The invention uses the constructed reverse genetics platform of ZJ1 strain of Newcastle disease virus of goose origin. The invention replaces two envelope glycoprotein gene fragments F and HN of an isolated strain JS-5-05-Go of Newcastle disease virus with high reproductive performance with the corresponding fragments of the ZJ1 strain of Newcastle disease virus of goose origin, so that the recombinant virus NDV-VII is obtained. The VII gene type of Newcastle disease virus A-NDV-VII which is highly attenuated is rescued after the attenuated mutation of the F gene of the recombinant virus. And the virushas a higher reproduction titer on chicken embryo. The invention is suitable for a mass production of vaccine, which can be used for the manufacture of vaccine.

Description

technical field [0001] The invention relates to the application of reverse genetic technology to produce a gene VII type Newcastle disease virus attenuated strain A-NDV-VII, which is used 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. A research technique for assembling new RNA viruses from genomic cDNA and various accessory proteins [Neumann G, Whitt M A, Kawaoka Y. Adecade after the generation of a negative-sense RNA virus from cloned cDNA-what have we learned? Journal of General Virology, 2002, 83(11): 2635-2662.], also known as full-length infectious cDNA cloning technology, is often referred to as "virus rescue". Since the "rescued" RNA virus is derived from cDNA clones, va...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/01C12N7/04
Inventor 刘秀梵胡顺林孙庆吴艳涛刘文博
Owner YANGZHOU UNIV