[Cloning, expression, purification and characterization of human endostatin gene].
OBJECTIVE
To procure biologically active human endostatin.


METHODS
Human endostatin gene was acquired by means of reverse transcriptase (RT)-PCR and cloned into PGEM-T vector with subsequent sequence identification. The gene fragment was inserted into the prokaryotic expression vector pBV220 and transformed into E.coli DH5alpha strain. Endostatin expression in the E.coli was identified and the inclusion body isolated, purified and its activity analyzed.


RESULTS
The obtained gene fragment 552 bp in length was identified as the functional section of human endostatin gene by sequence analysis, and SDS-PAGE analysis showed that the expressed product was the target protein with biological activity.


CONCLUSIONS
Human endostatin gene was expressed in E.coli and the protein obtained can inhibit the proliferation of ECV 304 cells.
Yang
Fang
F
Department of Obstetrics and Gynecology, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China.
He
Yuan-Li
YL
Jiang
Xiao-Yu
XY
Liu
Yun
Y
Peng
Dong-Xian
DX
Zong
Li-Li
LL
D003001:Cloning, Molecular; D043169:Endostatins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005822:Genetic Vectors; D006801:Humans; D014162:Transfection; D014471:Umbilical Veins / Q000166:cytology
D043169:Endostatins