Construction and application of mammal cell high-efficiency expression vector

A eukaryotic expression vector and vector technology, which can be applied to cells modified by introducing foreign genetic material, using vectors to introduce foreign genetic material, recombinant DNA technology, etc. Many other problems, to achieve the effect of reducing low expression background clones, overcoming position effects, and improving efficiency
CN103525868AActive Publication Date: 2014-01-22BIOTECH PHARMA CO LTD
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
BIOTECH PHARMA CO LTD
Publication Date
2014-01-22

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Abstract

The invention relates to a mammal cell high-efficiency expression vector pSNEO. The vector is constructed on the basis of neomycin resistance screening gene NEO by combined strategy of weakening screening gene expression and reinforcing target gene expression. The screening gene weakening comprises the following two sides: introducing deletion mutant into the promoter to implement transcription weakening of the screening gene, and introducing a hairpin structure sequence before the translation initial site of the screening gene to implement the translation weakening of the screening gene. The expression reinforcement of the target gene is implemented by adding a transcription control element WPRE sequence between polyclone site and polyA tail of the target gene expression frame. The pSNEO vector can conveniently and quickly complete the construction of the stable high-expression cell strain of the target gene, and provides a new tool for preparing the high-polymer recombinant protein.
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Description

technical field

[0001] The invention relates to biopharmaceutical gene engineering expression technology, in particular to the construction and application of a mammalian cell high-efficiency expression vector. Background technique

[0002] The acquisition of high-efficiency mammalian cell expression system and high-expression engineered cell line is the bottleneck in the research and production of genetically engineered drugs, and expression vector and host cell are the two basic components of mammalian cell expression system. The expression systems currently used to prepare genetically engineered drugs include prokaryotic expression systems with Escherichia coli as hosts and eukaryotic expression systems with yeast, insect cells, plant cells and mammalian cells as hosts.

[0003] The CHO expression system is one of the most widely used eukaryotic expression systems and one of the few animal cells that can be cultured in high-density suspension. The exogenous gene product ...

Claims

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