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Bombyx mori nuclear polyhydrosis virus (BmNPV) 39k inducible promoter and application thereof

A karyotype polyhedron, inducible technology, applied in the direction of using vectors to introduce foreign genetic material, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems that have not yet been reported on the BmNPV inducible promoter, and achieve inhibition of virus proliferation. Effect

Inactive Publication Date: 2010-12-15
SOUTHWEST UNIVERSITY
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Problems solved by technology

But so far, there is no research report on the BmNPV-inducible promoter

Method used

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  • Bombyx mori nuclear polyhydrosis virus (BmNPV) 39k inducible promoter and application thereof
  • Bombyx mori nuclear polyhydrosis virus (BmNPV) 39k inducible promoter and application thereof
  • Bombyx mori nuclear polyhydrosis virus (BmNPV) 39k inducible promoter and application thereof

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Embodiment Construction

[0020] The preferred embodiments of the present invention will be described in detail below with reference to the accompanying drawings. The experimental methods that do not specify specific conditions in the preferred embodiments usually follow conventional conditions, such as those described in the molecular cloning experiment guide (third edition, J. Sambrook et al., translated by Huang Peitang et al., Science Press, 2002) Or in accordance with the conditions recommended by the manufacturer.

[0021] 1. Selection and cloning of BmNPV inducible promoter

[0022] Download the BmNPV whole genome sequence (NC001962.1) from the National Center for Biotechnology Information (NCBI). According to the cascade regulation characteristics of baculovirus, the delayed early gene promoter (β gene) is affected by the immediate early gene (α gene) product To activate the regulation, select the 39k promoter of BmNPV delayed early gene as a candidate promoter.

[0023] The following PCR-specific p...

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Abstract

The invention discloses a bombyx mori nuclear polyhydrosis virus (BmNPV) 39k inducible promoter and application thereof. A nucleotide sequence of the promoter is shown as SEQ ID No.1; the promoter has obvious BmNPV inducible promotion activity and can make cells start foreign gene expression when the cells are subjected to the BmNPV infection; an RNAi vector using a BmNPV multiplication essential gene as a target can be built by utilizing the promoter; the abundant expression of shRNA can be performed when the bombyx mori is subjected to the BmNPV infection; the shRNA is cut into siRNA in a cell by an enzyme; specific degradation is performed on the BmNPV multiplication essential gene mRNA to initiate the BmNPV multiplication essential gene to be transcribed and then silenced, so the aim of inhibiting the virus multiplication is fulfilled, and the promoter can be used for preventing and controlling the BmNPV infection of the bombyx mori, performing genetic engineering breeding of bombyx mori anti-BmNPV strains and providing a good reference mode for transgenic therapy of other biologic virus diseases.

Description

Technical field [0001] The present invention relates to the field of biology, in particular to the Bombyx mori nuclear polyhedrosis virus (BmNPV) 39k inducible promoter and its application. Background technique [0002] Silkworm is a lepidopteran insect with important economic value. BmNPV is the most common and most harmful pathogen in sericulture production and has a huge impact on sericulture. How to use today's rapidly developing genetic engineering technology to fundamentally improve silkworm virus resistance is an important research topic in the field of sericulture. Gene knockout studies have shown that BmNPV will not be able to proliferate without certain genes necessary for proliferation. Therefore, inhibiting the expression of genes necessary for the proliferation of these viruses will become an effective strategy to prevent BmNPV infection in silkworms. [0003] RNA interference (RNAi) refers to the phenomenon of post-transcriptional gene silencing caused by the degra...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/63C12N1/15C12N1/19C12N1/21C12N5/10A01K67/04
Inventor 潘敏慧鲁成李娜赵爱春夏庆友向仲怀
Owner SOUTHWEST UNIVERSITY
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