Glutamine synthetase gene and application

A glutamine and synthetase technology, applied in the direction of enzymes, ligases, and the introduction of foreign genetic material using vectors, can solve the problems of difficulty in screening high-expressing cell lines, and the expression efficiency cannot meet production requirements, and achieve the effect of improving expression.

Inactive Publication Date: 2019-07-09
GENSCRIPT NANJING
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  • Abstract
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Problems solved by technology

[0005] However, the existing GS system still has the problems that the expression efficiency cannot meet the production requirements and the high expression cell lines are difficult to screen

Method used

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  • Glutamine synthetase gene and application
  • Glutamine synthetase gene and application
  • Glutamine synthetase gene and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0028] 1. Construction of recombinant expression vector

[0029] The light chain nucleotide sequence of antibody mAb01 is shown in SEQ ID NO:1, and the heavy chain nucleotide sequence is shown in SEQ ID NO:2. The light chain nucleotide sequence of the antibody mAb02 is shown in SEQ ID NO:3, and the heavy chain nucleotide sequence is shown in SEQ ID NO:4. The light chain nucleotide sequence of antibody mAb03 is shown in SEQ ID NO:5, and the heavy chain nucleotide sequence is shown in SEQ ID NO:6.

[0030] The eukaryotic expression vector containing the GS16034 gene and the target antibody sequence can be constructed by the following steps: (1) replace the sequence between the 2136-2931bp of the initial vector pcDNA3.1 (+) site with mGS to obtain the intermediate vector pcDNA3.1-mGS, and then The initial vector pcDNA3.1 (+) site 232-819bp sequence was cloned into the intermediate vector pcDNA3.1-mGS between the 1252-1253bp site to obtain a new vector pcDNA3.1-mGS-DGV, and the v...

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Abstract

The invention provides a new glutamine synthetase gene having a nucleotide sequence shown as SEQ ID NO: 8 and further provides a eukaryotic expression vector containing the same. The glutamine synthetase gene can be used in a GS gene amplification system to promote expression of target protein and screening of high-expression cell lines.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a novel glutamine synthetase gene and the application of the glutamine synthetase gene in the construction of eukaryotic expression vectors. Background technique [0002] According to the time difference of the expression of the target protein and whether the expression process needs to add screening pressure, the expression system can be divided into transient expression system and stable expression system. The transient expression system refers to that the host cells do not add screening pressure after introducing the expression vector containing the foreign gene, and collect the protein produced by the cell. The transfected cells gradually lose the target protein as they divide, and cannot continue to produce protein for a long time. The stable expression system means that the expression vector enters the host cell and is selected and cultivated by screening pressure. ...

Claims

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Application Information

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IPC IPC(8): C12N15/52C12N9/00C12N15/85C12N15/66
CPCC12N9/93C12Y603/01002C12N15/85C12N15/66C07K14/47C12N9/00C12N15/52C12N15/69
Inventor 时红星赵钰陈明月祁碧玉贺伟伟邹晋晋张丽华
Owner GENSCRIPT NANJING
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