[Gene cloning and prokaryotic expression of the PID domain of human DOC-2 molecule].
OBJECTIVE
To clone the PID domain of human DOC-2 (nDOC-2) and express it in E.coli DH5alpha.


METHODS
The cDNA fragment encoding the PID domain of nDOC-2 was amplified by RT-PCR from normal human ovarian tissue and cloned to pUC19. The DNA fragment from the pUC19-nDOC-2 digested with BamH I and EcoR I was ligated to the BamH I/EcoR I digested prokaryotic expression vector pGEX-4T-1.The expression of fusion protein was induced with IPTG and the expressed product was identified by SDS-PAGE.


RESULTS
(1)The sequencing and endonucleases digestion analysis showed that the fragment of nDOC-2 gene was insert into vectors pUC19 and pGEX-4T-1;(2)SDS-PAGE showed the nDOC-2 gene had been expressed in E.coli DH5alpha.


CONCLUSIONS
The PID domain of nDOC-2 was expressed successfully in prokaryote, which makes preparation for further researching the function of DOC-2 and preparing antibodies to DOC-2 protein.
Liu
Shu-Juan
SJ
Department of Obstetrics and Gynecology,Xijing Hospital, Fourth Military Medical University, Xi'an 710032 China.
Zheng
Wei-Guo
WG
Ji
Zong-Ling
ZL
Chen
Su-Min
SM
Zhang
Xin-Hai
XH
Yang
Li-Jun
LJ
D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D010957:Plasmids