Construction and application of vector used for developing high-expression cell strain

A eukaryotic expression vector and vector technology, which can be applied in the direction of using vectors to introduce foreign genetic material, cells modified by the introduction of foreign genetic material, and recombinant DNA technology. The effect of overcoming the position effect and improving the efficiency
CN103525867AActive Publication Date: 2014-01-22BEIJING DONGFANG BIOTECH
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Patent Information

Authority / Receiving Office
CN ยท China
Current Assignee / Owner
BEIJING DONGFANG BIOTECH
Publication Date
2014-01-22

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Abstract

The invention relates to a pSGS vector and application of the vector to development of a high-expression cell strain. The construction of the vector comprises the following steps: based on a neomycin resistance screening gene NEO, through combined strategy of weakening screening gene expression and reinforcing target gene expression, constructing an expression vector pSNEO, and in view of the amplification function of a GS system, through designing a primer, replacing the hairpin structure sequence and the NEO gene of the vector pSNEO with a hairpin structure sequence and a GS gene to obtain a universal vector pSGS for screening a high-expression and stable-transfection cell strain. Through the pSGS vector constructured by the invention, the construction of a stable and high-expression cell strain of the target gene can be finished conveniently and quickly, and a new tool is provided for preparation of macromolecular recombinant protein.
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Description

technical field

[0001] The invention relates to the gene engineering expression technology of biopharmaceuticals, in particular to the construction and application of a vector used for the development of high-expression cell lines. 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 the main host cell 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 suspen...

Claims

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