[Cloning of human obesity gene and its expression in E. coli].
OBJECTIVE
To clone the obesity gene of Chinese and express human leptin in E.coli.


METHODS
The obesity gene was amplified from the total RNA isolated from cultured human adipocytes of Chinese by reverse transcriptional PCR, inserted into TA-vector and cloned into the expression plasmid pBV220 after sequence identification.


RESULTS
DNA sequencing confirmed that the isolated obesity gene was identical to the previously reported sequence. The recombinant plasmid pBV220-OB was constructed and leptin successfully expressed in E.coli.


CONCLUSIONS
Successful cloning and expression of human obesity gene in E.coli may facilitate further research of the mechanism of fat metabolism and adipocyte differentiation.
Zhu
Ji
J
Institute of Molecular Biology, Southern Medical University, Guangzhou 510515, China.
Ma
Wen-Li
WL
Mao
Xiang-Ming
XM
Li
Ling
L
Song
Yan-Bin
YB
Zheng
Wen-Ling
WL
D017667:Adipocytes / Q000166:cytology* / Q000378:metabolism; D001483:Base Sequence; D002478:Cells, Cultured; D018076:DNA, Complementary / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D020738:Leptin / Q000096:biosynthesis* / Q000235:genetics; D008969:Molecular Sequence Data; D009765:Obesity / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA
D018076:DNA, Complementary; D020738:Leptin