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[Gene cloning and expression of CTLA-4 in E. coli]
1Department of Bioengineering, Institute of Medicinal Biotechnology, CAMS, PUMC, Beijing 100050, China.
Objective:
To express hsCTLA-4 in E. coli.
Methods:
The hsCTLA-4 gene was obtained by PCR amplification from pE plasmid which contains CTLA-4 gene and was inserted into the expression vector pGEX-2T. The recombination strain was induced by IPTG with different concentrations and time.
Results:
The sequence of PCR amplified DNA fragments was identical with the reported CTLA-4 gene. SDS-PAGE and Western blot showed that 0.10 mmol/L IPTG can induce higher production of the fusion protein with molecular weight 40,000 after addition of IPTG 4 hours and GST-CTLA4 had immunological activity.
Conclusions:
hsCTLA-4 can be expressed in soluble form high efficiency.