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Rhabdovirus expression vector capable of relieving high-titer inhibition

A baculovirus and expression vector technology, applied in the field of baculovirus expression vector, can solve the problem that the baculovirus expression system does not have gene dose effect and the like

Active Publication Date: 2020-07-21
陕西杆粒生物科技有限公司
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  • Abstract
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  • Claims
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Problems solved by technology

[0007] The purpose of the present invention is to provide a baculovirus vector that relieves high-titer inhibition, aiming to solve the problem that the baculovirus expression system mentioned in the background technology does not have a gene dosage effect

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  • Rhabdovirus expression vector capable of relieving high-titer inhibition
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  • Rhabdovirus expression vector capable of relieving high-titer inhibition

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Embodiment Construction

[0016] 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.

[0017] The application principle of the present invention will be described in detail below in conjunction with the accompanying drawings (taking the replacement of alanine as an example).

[0018] 1. Knockout of leucine at position 21 of LEF-10 on Bacmid

[0019] Use primers (SEQ ID NO:1 and SEQ ID NO:2) with 50bp homology arms upstream and downstream of the 21st site of LEF-10, and use the prpsL-AMP plasmid as a template to amplify rpsL-AMP (ampicillin-resistant gene) fragment, a 1023bp PCR product was obtained.

[0020] The obtained PCR product was transformed into E. coli strain HS996 with RedET plasmid and Bacmid by ...

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Abstract

The invention discloses a rhabdovirus expression vector capable of relieving high-titer inhibition. A rhabdovirus expression system does not have a significant dose effect, that is, the high virus titer cannot increase the yield of exogenous genes, because the high titer induces aggregation of later-period expression factors LEF-10, and then expression of later-period genes is inhibited. Accordingto the vector, through mutation of the 21st leucine site of the LEF-10, aggregation tendency of the LEF-10 is lowered. Insect cells are infected by high-titer mutation rhabdovirus, thus the expression amount, controlled by later-period promoters, of the exogenous genes can be significantly increased, and through increase of the expression yield, the production cost of enterprises can be significantly lowered. The rhabdovirus expression vector can be used in the field of biological product industry, especially the field of subunit vaccine industry.

Description

technical field [0001] The invention belongs to the technical field of recombinant protein expression, and in particular relates to a baculovirus expression vector capable of releasing high-titer inhibition. Background technique [0002] Baculovirus is a double-stranded DNA virus that specifically infects arthropods. Autographa californica nucleopolyhedrovirus (AcMNPV) is the type species of baculovirus. Since Smith GE et al first expressed human β-interferon gene in insect cells with baculovirus in 1983 (Mol Cell Biol. 1983; 3:2156-65), due to its low cost, high yield, and various post-translational Modification system and other advantages, the baculovirus expression vector system is widely used in scientific research and production. [0003] In the field of protein expression, it is recognized that gene expression has a dosage effect, that is, there is a certain positive correlation between gene copy number and protein expression, but the baculovirus expression system is ...

Claims

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Application Information

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IPC IPC(8): C12N15/866
CPCC12N15/86C12N2710/14043
Inventor 不公告发明人
Owner 陕西杆粒生物科技有限公司
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