High-infection quasi-influenza virus preparation as well as special coding sequence and application thereof

An influenza virus, special coding technology, applied in the field of biotechnology inventions

CN102234635AInactive Publication Date: 2011-11-09中国疾病预防控制中心病毒病预防控制所
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Publication Date
2011-11-09
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention particularly relates to a high-infection quasi-influenza virus system which is mainly characterized in that carriers of LTR(Long Terminal Repeat) containing HIV(Human Immunodeficiency Virus)-1, promoters, gag-pol and other elements and high-infection influenza virus membrane protein expression plasmids are subjected to co-transfection and packaging to form quasi-influenza viruses which have infection characteristics and antigenicity of high-infection influenza virus membrane protein. The system is used as a convenient and safe platform for researches on the physiological characteristics of highly pathogenic influenza virus membrane protein and researches on neutralizing antibodies. The system is used as an efficient gene introduction carrier, a target gene can be inserted into a gene-containing system reconstructed through packaging signals of the HIV-1, and the target gene can be introduced into various cells through quasi-virus infection according to retrovirus integration rules. After the cells are infected by quasi-viruses, reporter genes are integrated and expressed through gag-pol protein of the HIV-1, and the inhibition to the gag-pol function by medicaments can be reflected through the expression of the reporter genes, thus the system can be used as an evaluation platform for gag-pol medicaments.
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Description

Technical field

[0001] The invention belongs to the field of biotechnology inventions. The present invention specifically relates to a high infectivity pseudo-influenza virus system, which is mainly co-transfected and packaged by a vector containing human immunodeficiency virus LTR, promoter, gag-pol and other elements and a plasmid containing high infectivity avian influenza virus membrane protein The virus-like system has the characteristics of highly infectious avian influenza virus; the highly pathogenic avian influenza membrane proteins (including hemagglutinin HA and neuraminidase) present on the surface of the virus-like virus NA) has the same antigenicity as the ancestor virus, so it is a convenient and safe platform for the study of the physiological characteristics of the highly pathogenic avian influenza virus membrane protein and the study of neutralizing antibodies; the system can be used as a highly efficient gene transfer vector through human immunodeficiency viru...

Examples

Embodiment Construction

[0011] 1. On the basis of the existing lentiviral system for packaging pseudoviral particles, our laboratory has improved the original lentiviral packaging system, adding a multiple cloning site MCS downstream of the reporter gene in the transfer plasmid with the reporter system, The plasmid can insert the target sequence downstream of the reporter gene EGFP, named pTrs-EGFP-v, inserting a specific gene can introduce the target gene into a variety of cells through pseudo-virus infection, and the introduced gene can be realized according to the retrovirus integration law Efficient expression of single or multiple genes.

[0012] 2. Synthesize multiple strains of influenza virus membrane protein expression genes HA and NA, respectively clone them into high-efficiency expression protein vector pWY for compatibility combination, and verify their high-efficiency expression in virus-like packaging cells 293T. pWY is our own plasmid vector, which is used for high-efficiency expression o...