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APMV (avian paramyxovirus) fusion protein, preparation method and application thereof and APMV vaccine for pigeons

A technology of avian paramyxovirus and fusion protein, applied in the direction of antiviral immunoglobulin, veterinary vaccine, biochemical equipment and methods, etc., can solve the disadvantages of emergency vaccination and reduce the cost of vaccines, have potential infection risks, and are difficult to promote Application and other issues to achieve the effect of preventing the prevalence and spread of infection, good antigenicity, and reducing morbidity and mortality

Active Publication Date: 2019-04-09
天康生物制药有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although inactivated vaccines are safe and stable, they require large doses of vaccination or the application of concentrated antigens; the immunity period is short, and booster vaccination is often required; it takes 2 weeks to produce complete immunity, which is not conducive to emergency vaccination and reduced vaccine costs, and the protective effect is not good
However, attenuated virus vaccines are difficult to popularize and apply in practice due to defects such as high cost, easy reversion to ancestors, and potential risk of infection.

Method used

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  • APMV (avian paramyxovirus) fusion protein, preparation method and application thereof and APMV vaccine for pigeons
  • APMV (avian paramyxovirus) fusion protein, preparation method and application thereof and APMV vaccine for pigeons
  • APMV (avian paramyxovirus) fusion protein, preparation method and application thereof and APMV vaccine for pigeons

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Experimental program
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preparation example Construction

[0051] The present invention also provides a kind of above-mentioned avian paramyxovirus fusion protein, the preparation method of above-mentioned avian paramyxovirus fusion protein or the application of the avian paramyxovirus fusion protein prepared by the preparation method, including the following (x1)-(x6 ) in at least one: (x1) preparation of pigeon subunit dual vaccine; (x2) preparation of vaccine for poultry viral diseases; (x3) preparation of antibody to avian paramyxovirus; (x4) preparation of antigen of avian paramyxovirus (x5) preparing reagents and / or kits for detecting avian paramyxovirus; (x6) preparing reagents and / or kits for detecting avian paramyxovirus antibodies.

[0052] The avian paramyxovirus fusion protein provided by the present invention expresses the HN gene of pigeon type 1 paramyxovirus, that is, Newcastle disease virus and avian paramyxovirus type 7 in series, and the avian paramyxovirus fusion protein not only contains pigeon type 1 paramyxovirus...

Embodiment 1

[0058] Embodiment 1 Expression of avian paramyxovirus fusion protein

[0059] 1.1 Synthesis and amplification of the target gene

[0060] Obtain the whole viral RNA sequence of NDV and APMV-7 through GenBank, and find the DNA sequence encoding the HA gene, wherein the amino acid sequence of the HN gene encoding NDV is shown in SEQ ID NO.1, and the nucleotide sequence is shown in SEQ ID NO.4 shown; the amino acid sequence of the HN gene encoding AMPV-7 is shown in SEQ ID NO.2, and the nucleotide sequence is shown in SEQ ID NO.5.

[0061] After codon optimization of the gene sequence according to the expression host, a flexible fusion protein linking peptide (PPSPS) was selected 2 The HN gene of NDV and the HN gene of APMV-7 were connected, and the sequence was named NDAP-HN, and its amino acid sequence was shown in SEQ ID NO.3, and was sent to a biological company for synthesis.

[0062] 1.2 Expression vector construction

[0063] The synthetic NDAP-HN gene and pMG36e expres...

Embodiment 2

[0076] The preparation of embodiment 2 vaccine

[0077] Clean the vials and wrap them up, sterilize at 121°C for 15 minutes, and dry them for later use. Multi-valve vial stoppers ready to be sterilized. Inoculate the activated single colony into the test tube containing the culture medium, and after cultivation, add the freeze-dried protective agent (20g milk powder + 80g pure water) and mix evenly at a ratio of 1:3; add 2ml of the mixed solution to each bottle and cover it Cork (from the gap between the cork flaps), with the strain number written on the bottle. Store the aliquoted mixture in a -20°C refrigerator overnight. The next day, the mixed solution was frozen in a -80°C refrigerator for 2 h. Then put the vials containing the mixed solution together into a freeze dryer for extraction and pre-freeze drying. Freeze-dry according to the operating procedures of the freeze-dryer. Finally, the vial was taken out, capped, and the effect of freeze-drying was observed, and ...

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Abstract

The invention provides APMV (avian paramyxovirus) fusion protein, a preparation method and an application thereof and an APMV vaccine for pigeons, and relates to the technical field of biology. The APMV fusion protein is obtained by serial connection of an HN gene of PPMV-1 (pigeon paramyxovirus-1) and an HN gene of APMV-7, and comprises an NDV-HN segment and an AMPV-7-HN segment, wherein the NDV-HN segment comprises the amino acid sequence shown in SEQ ID NO.1, and the AMPV-7-HN segment comprises the amino acid sequence shown in SEQ ID NO.2. The APMV fusion protein has the advantages of goodantigenicity, high expression quantity and wide application.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an avian paramyxovirus fusion protein, its preparation method, application and APMV vaccine for pigeons. Background technique [0002] Recently, there have been many large-scale pigeon farms with tens of thousands to tens of thousands of pairs, and some with 100,000 to 200,000 pairs. Only in Guangdong Province, the annual stock of pigeons has reached more than 4 million pairs, and the production of squabs is more than 45-50 million. It is a large province for raising pigeons in China, and the breeding volume in other provinces and cities is about 3-4 times that of Guangdong. Among them, Jiangsu, Shandong, Hebei, Hunan, Guangxi, and southern Xinjiang and other places are more breeding. Based on such a large-scale breeding environment, it brings huge profits to the pigeon industry. [0003] However, in the process of rapid development of meat pigeon breeding, the occurrence and preval...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/74C07K16/10A61K39/17A61P31/14
CPCA61K39/12A61K2039/552A61K2039/70A61P31/14C07K14/005C07K16/1027C07K2319/00C12N15/74C12N2760/18022C12N2760/18122C12N2760/18134
Inventor 贺笋任芳赵毅刘宏张丹李森江叶青
Owner 天康生物制药有限公司
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