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Method of saving chimeric viral strain and chimeric attenuated strain with A/B type chimeric HA plasmids

A chimeric virus and plasmid technology, applied in the biological field, can solve the problems of attenuated live vaccines such as virulence reversion, and achieve the effect of solving virulence reversion, strong immunogenicity and high safety

Inactive Publication Date: 2019-06-21
ACAD OF MILITARY SCI PLA CHINA ACAD OF MILITARY MEDICAL SCI INST OF MILITARY VETERINARY MEDICINE
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Problems solved by technology

[0004] The purpose of the present invention is to provide a chimeric virus strain containing A / B type chimeric HA plasmid and a chimeric attenuated strain rescue method to solve the problem that current attenuated live vaccines are prone to virulence reversion due to vaccine strains

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  • Method of saving chimeric viral strain and chimeric attenuated strain with A/B type chimeric HA plasmids
  • Method of saving chimeric viral strain and chimeric attenuated strain with A/B type chimeric HA plasmids
  • Method of saving chimeric viral strain and chimeric attenuated strain with A/B type chimeric HA plasmids

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Embodiment Construction

[0064] The following will clearly and completely describe the technical solutions in the embodiments of the present invention with reference to the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only some, not all, embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without making creative efforts belong to the protection scope of the present invention.

[0065] The construction method of A / B type chimeric HA plasmid, concrete steps are as follows:

[0066] Step S1, gene synthesis: modify the alkaline cleavage site of the HA sequence of H5 subtype influenza 2.3.4.4 lineage A / cat / Sichuan / SC18 / 2014, such as image 3 As shown, the three basic amino acids near the basic cleavage site were removed, and the three amino acid codons near it were changed, that is, the three basic amino acids from the 342nd to the 344th ...

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Abstract

The invention disclose a method of saving a chimeric viral strain and chimeric attenuated strain with A / B type chimeric HA plasmids. The method comprises: subjecting an HA sequence of H5 subtype influenza 2.3.4.4 lineage to alkaline cleavage site modification such that a highly pathogenic virus is converted into a lowly pathogenic virus; synthesizing an HA gene of the H5 subtype influenza 2.3.4.4lineage; designing and synthesizing an amplification primer to construct A / B type chimeric HA plasmids by using the synthesized HA gene and constructed pBD-Ya1688-HA transflective plasmids as templates; cloning the HA gene of H5 subtype influenza virus directionally to the pBD-Ya1688-HA transflective plasmids to form A / B type chimeric HA plasmids by means of seamless cloning; saving the chimeric viral strain and chimeric attenuated strain with A / B type chimeric HA plasmids through the A / B type chimeric HA plasmids. The problem is solved that an influenza virus vaccine easily causes virulent recovery.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a chimeric virus strain containing an A / B type chimeric HA plasmid expressing H5 subtype 2.3.4.4 lineage influenza virus HA protein and a chimeric attenuated strain rescue method. Background technique [0002] At present, there are mainly inactivated vaccines and cold-adapted attenuated vaccines in clinical practice. Inactivated vaccines can only cause humoral immunity, and regular immunization is required every year; cold-adapted attenuated vaccines can induce mucosal immunity, and the immune effect is good, but the safety is not as good as inactivated vaccines, because it is a live virus, and there is a risk of virulence returning to the ancestors. [0003] Inactivated vaccines and live attenuated vaccines induce different immune responses: inactivated vaccines are generally administered by intramuscular injection, mainly causing humoral immunity; live attenuated vaccines can be inoc...

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Application Information

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IPC IPC(8): C12N15/85C12N7/01C12N7/04C12R1/93
Inventor 高玉伟赵梦琳孙伟洋于志君王铁成冯娜赵永坤张雪梅杨松涛夏咸柱
Owner ACAD OF MILITARY SCI PLA CHINA ACAD OF MILITARY MEDICAL SCI INST OF MILITARY VETERINARY MEDICINE
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