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Recombination bollworm monokaryon capsid polyhedrosis virus for expression polghedrosis and chitinase fusion protein and its preparation method

A nuclear polyhedrosis virus and chitinase technology, applied in the field of microorganisms, can solve the problems of late expression time and slow onset of exogenous proteins, and achieve the effects of improving efficiency and increasing insecticidal speed.

Inactive Publication Date: 2003-07-16
WUHAN INST OF VIROLOGY CHINESE ACADEMY OF SCI
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Problems solved by technology

Solved the problem of late expression of exogenous protein and slow onset of the existing recombinant virus insecticides

Method used

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  • Recombination bollworm monokaryon capsid polyhedrosis virus for expression polghedrosis and chitinase fusion protein and its preparation method

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Embodiment Construction

[0009] 1. The recombinant Helicoverpa armigera mononuclear capsid nuclear polyhedrosis virus HaSNPV-WD6 provided by the present invention. The egt gene, the ecdysone uridine diphosphate glucosyltransferase gene, is deleted from its genome. At the site where the ecdysone uridine diphosphate glucosyl transferase gene is deleted, the virus’s own polyhedrin gene promoter is inserted. Polyhedrin and chitinase fusion gene. And the marker gene controlled by the promoter of the heat shock protein gene (hsp70) is the β-galactosidase gene (LacZ).

[0010] The above chitinase gene can also be derived from other genes with the same effect, such as the cathepsin gene v-cath of baculovirus, gp37 gene, the spindle protein gene fusolin of insectpox virus, and the Bacillus thuringiensis insecticidal crystal protein (IAP). ) Gene replacement. The insertion site of the fusion gene can be at the egt site or at other sites in the viral genome. The marker gene can also be replaced with the green fluore...

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Abstract

A recombinant mononuclear capsid nucleopolyhedrovirus of bollw orm able to express polyhedron and chitinase fusion protein is prepared through PCR amplification of polyhedron gene HaSNPV and chitinase gene, cloning to carrier pUC19 to configure fusion gene, inserted it in eukaryotic carrier pHaWHL5 to obtain eukaryotic transfer carrier pHaWD6, transfecting insection cell by it along with wild virus DNA, and homologous recombination.

Description

Technical field [0001] The present invention relates to the field of microorganisms, in particular to a genetically recombinant cotton bollworm mononuclear capsid nuclear polyhedrosis virus capable of controlling cotton bollworm and tobacco budworm; the present invention also relates to the preparation of the recombinant cotton bollworm mononuclear capsid nuclear polyhedrosis virus strain method. Background technique [0002] Baculovirus has been widely used in the biological control of agricultural and forestry pests since the 1970s. At the same time, the baculovirus-insect (cell) system is still a very good eukaryotic expression system, which has been used in the research and production of foreign genes, diagnostic reagents and vaccines. Like other baculoviruses, Helicoverpa armigera single nucleocapsidnucleopolyhedrovirus (HaSNPV) as a biological insecticide has the disadvantage of slow insecticidal speed, which limits its large-scale application. A variety of recombinant viru...

Claims

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Application Information

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IPC IPC(8): A01N63/00C12N7/01C12N15/33C12N15/62C12N15/79
Inventor 吴东陈新文胡志红
Owner WUHAN INST OF VIROLOGY CHINESE ACADEMY OF SCI
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