[Cloning of PD-1 gene and its prokaryotic expression in Escherichia coli].
OBJECTIVE
To clone human PD-1 gene, construct a prokaryotic expression plasmid and express in E. coli.


METHODS
The 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.


RESULTS
The 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.


CONCLUSIONS
The 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.
Chen
Si-yong
SY
Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China.
Guan
Kun-ping
KP
Guo
Min-zhuo
MZ
Yi
Yao
Y
Jia
Zhi-yuan
ZY
Yu
Tao
T
Guo
Yu
Y
Bi
Sheng-li
SL
D000595:Amino Acid Sequence; D015703:Antigens, CD / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D051017:Apoptosis Regulatory Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D006801:Humans; D016133:Polymerase Chain Reaction; D061026:Programmed Cell Death 1 Receptor; D011387:Prokaryotic Cells / Q000378:metabolism*; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
D015703:Antigens, CD; D051017:Apoptosis Regulatory Proteins; C105992:PDCD1 protein, human; D061026:Programmed Cell Death 1 Receptor