[Gene cloning and expression of CTLA-4 in E. coli].
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.
Wang
C J
CJ
Department of Bioengineering, Institute of Medicinal Biotechnology, CAMS, PUMC, Beijing 100050, China.
Li
Y
Y
D015703:Antigens, CD; D000943:Antigens, Differentiation / Q000096:biosynthesis* / Q000235:genetics; D060908:CTLA-4 Antigen; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics
D015703:Antigens, CD; D000943:Antigens, Differentiation; D060908:CTLA-4 Antigen; D011994:Recombinant Proteins