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Recombined Newcastle disease heat-resisting vaccine strain for expressing H5 subtype avian influenza virus truncated HA protein and preparation method

An avian influenza virus and vaccine strain technology, applied in the field of molecular biology, can solve the problem of lack of heat resistance, and achieve the effects of saving storage and transportation costs, reducing dependence, and significantly heat resistance.

Inactive Publication Date: 2016-09-21
INST OF ANIMAL SCI & VETERINARY HUBEI ACADEMY OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The vaccine strain does not have significant heat tolerance and expresses the complete HA protein

Method used

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  • Recombined Newcastle disease heat-resisting vaccine strain for expressing H5 subtype avian influenza virus truncated HA protein and preparation method
  • Recombined Newcastle disease heat-resisting vaccine strain for expressing H5 subtype avian influenza virus truncated HA protein and preparation method

Examples

Experimental program
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Effect test

Embodiment 1

[0025] The first step, construction and identification of recombinant full-length plasmid pTS-HA1 / H5

[0026] 1.1 PCR amplification of HA1 gene of H5 subtype avian influenza virus

[0027] H5N1 avian influenza virus A / chicken / Hubei / 489 / 2004 strain (this virus strain has been reported in the literature, Luo Mengcheng, Prokaryotic expression and antibody preparation of H5N1 subtype avian influenza virus nucleoprotein NP, Chinese Journal of Zoonoses, 2008) proliferated on 9-11-day-old chicken embryos, and harvested chicken embryo allantoic fluid 5 days after inoculation. The harvested allantoic fluid was subjected to RT-PCR amplification of the HA1 gene, and the amplification primers were: upstream primer 5'- AAGCTTGCCACCATG GAGAAAATAGTGCTTCTTCTTGC-3', downstream primer 5'- ATCGGGGCACTCCGA TTCTACCCGTATTTTTCTTAATTATTATCCTCTCTTTTTTCTTCTTCTC-3' (the underlined part is the complementary sequence used for In-fusion cloning connection). The target band detected by agarose gel elec...

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Abstract

The invention discloses a recombined Newcastle disease heat-resisting vaccine strain for expressing H5 subtype avian influenza virus truncated HA protein and a preparation method. The heat-resisting vaccine strain is classified and named rTS-HA1 / H5, and is preserved in China Center for Type Culture Collection in Wuhan University, and the preservation number is CCTCC NO:V201629. The vaccine strain has remarkable heat-resisting performance, dependency on a cold chain system can be remarkably reduced, the preservation and transportation cost is reduced, and the heat stability of vaccines is improved. The incomplete HA protein is expressed by the vaccine strain, only an HA1 region (1-1041nt) with high immunogenicity in the HA protein is selected, and the immunoprotection effect of the vaccines can be enhanced specifically. The vaccine strain can be used for preparing an H5 avian influenza and Newcastle disease bigeminal heat-resisting vaccine.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to a recombinant Newcastle disease heat-resistant vaccine strain expressing truncated HA protein of H5 subtype avian influenza virus and a preparation method thereof. Background technique [0002] H5 subtype avian influenza virus can cause an acute fatal infectious disease of poultry - highly pathogenic avian influenza, its clinical manifestations are high mortality, decreased egg production, and severe respiratory symptoms. H5 subtype avian influenza can also infect humans and cause disease. In the "bird flu" incident in Hong Kong in 1997, the H5 subtype bird flu crossed the interspecies barrier and directly infected humans, causing 18 infections and 6 deaths. As of 2013, H5 subtype avian influenza has caused 648 infections and 384 deaths, including 45 infections and 30 deaths in my country. In addition to poultry, H5 subtype avian influenza strains have also been iso...

Claims

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Application Information

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IPC IPC(8): C12N7/01A61K39/295A61K39/145A61K39/17A61P31/14A61P31/16C12R1/93
CPCC12N7/00A61K39/12A61K2039/525A61K2039/552A61K2039/70C12N2760/16134C12N2760/18121C12N2760/18134C12N2760/18143A61K2300/00
Inventor 邵华斌温国元罗青平潘兹书乔磊罗玲王红琳张腾飞汪宏才张蓉蓉卢琴艾地云
Owner INST OF ANIMAL SCI & VETERINARY HUBEI ACADEMY OF AGRI SCI
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