Cloning of PD-1 gene and its prokaryotic expression in Escherichia coli.
- Author:
Si-yong CHEN
1
;
Kun-ping GUAN
;
Min-zhuo GUO
;
Yao YI
;
Zhi-yuan JIA
;
Tao YU
;
Yu GUO
;
Sheng-li BI
Author Information
- Publication Type:Journal Article
- MeSH: Amino Acid Sequence; Antigens, CD; biosynthesis; chemistry; genetics; isolation & purification; Apoptosis Regulatory Proteins; biosynthesis; chemistry; genetics; isolation & purification; Cloning, Molecular; Electrophoresis, Polyacrylamide Gel; Escherichia coli; genetics; Genetic Vectors; genetics; metabolism; Humans; Polymerase Chain Reaction; Programmed Cell Death 1 Receptor; Prokaryotic Cells; metabolism; Sequence Alignment; Sequence Analysis, DNA
- From: Chinese Journal of Experimental and Clinical Virology 2008;22(1):33-35
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo clone human PD-1 gene, construct a prokaryotic expression plasmid and express in E. coli.
METHODSThe human PD-1 cDNA was cloned by RT-PCR from the total RNA, which was extracted from peripheral blood lymphocyte cell of the patient with chronic hepatitis B. Recombinant PD-1 protein was been expressed and purified after the prokaryotic expression plasmid had been constructed. It was identified by SDS-PAGE, DNA sequencing and amino acid sequencing.
RESULTSThe PD-1 gene was cloned and confirmed by DNA sequencing. The recombinant protein was expressed in E. coli. The purified protein was obtained, then been confirmed by amino acid sequencing.
CONCLUSIONThe human PD-1 gene was successfully cloned and expressed in E. coli, which lays the foundation for further study on the function and application of PD-1.
