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Broad-spectrum anti-apoptosis rhabdovirus expression vector

A technology of baculovirus and expression vector, which is applied in the direction of antiviral agent, virus/bacteriophage, virus, etc., and can solve the problem of no baculovirus expression vector

Active Publication Date: 2018-11-02
陕西杆粒生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In industrial production, the two types of cells are usually used crosswise, but unfortunately, there is no baculovirus expression vector that can resist apoptosis in both Sf cells and Tn cells

Method used

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  • Broad-spectrum anti-apoptosis rhabdovirus expression vector
  • Broad-spectrum anti-apoptosis rhabdovirus expression vector
  • Broad-spectrum anti-apoptosis rhabdovirus expression vector

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Embodiment Construction

[0023] In order to make the object, technical solution and advantages of the present invention more clear, the present invention will be further described in detail below in conjunction with the examples. It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the present invention.

[0024] The application principle of the present invention will be described in detail below in conjunction with the accompanying drawings.

[0025] According to the consensus sequence SEQ ID NO:1 of Sf-caspase-1 and Tn-caspase-1, the present invention designs two target sites of RNAi:

[0026] gccgcactgagacagatggct (563-583)

[0027] gcactgagacagatggctcac (566-586)

[0028] Design four siRNA coding sequences according to these two target sites (DNA sequences are: SEQ ID NO:4 + loop +SEQ ID NO:2 + TTTTT and SEQ ID NO:5 + loop + SEQ ID NO:3 + TTTTT ):

[0029] 563-4T: gctgtattgagatagatggctgcttattaaagccatctgtctcagtgcggcttt...

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Abstract

The invention discloses a broad-spectrum anti-apoptosis rhabdovirus expression vector, and the broad-spectrum anti-apoptosis effect is achieved through the expression of the caspase-1 consensus sequence of target Spodoptera frugiperda and Trichoplusia ni insect cells by the vector. The rhabdovirus vector comprises a section of specific DNA sequence, the DNA sequence comprises the siRNA sequence ofthe consensus sequence of the target Sf-caspase-1 and Tn-caspase-1 transcribed by an RNA polymerase III promoter; the recombinant virus of the vector expresses double-strand small RNA in a host cell,the caspase-1 encoded by the silent host cell is interfered through the RNA, so that the apoptosis of the host cell is inhibited, and the expression level of a foreign protein is obviously improved.The broad-spectrum anti-apoptosis rhabdovirus expression vector can be used in the industrial production of protein preparations and vaccines.

Description

technical field [0001] The invention belongs to the technical field of recombinant protein expression, in particular to a baculovirus expression vector capable of resisting apoptosis in various insect cells. Background technique [0002] Baculovirus is a double-stranded DNA virus that specifically infects arthropods, Autographa californica polynuclear polyhedrosis virus ( Autographa californica nucleopolyhedrovirus, AcMNPV) is the type species of baculovirus. Since Smith GE and others first used baculovirus to express human β-interferon gene in insect cells in 1983, due to the advantages of low cost, high yield, and various post-translational modification systems, the baculovirus expression vector system has been used in many fields. It is widely used in scientific research and production. [0003] The baculovirus expression system is a transient expression system, and insect cells will undergo apoptosis 3-4 days after being infected by the virus. The expression time lim...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/866C12N15/113A61K39/12A61P31/20
CPCA61K39/12A61P31/20C12N15/86C12N2310/14C12N2710/14043C12N2710/14034C12N15/1131C07K14/005C12N2710/14022Y02A50/30
Inventor 不公告发明人
Owner 陕西杆粒生物科技有限公司
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