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[Cloning of human obesity gene and its expression in E. coli]
Ji Zhu1, Wen-Li Ma, Xiang-Ming Mao
1Institute of Molecular Biology, Southern Medical University, Guangzhou 510515, China.
Objective:
To clone the obesity gene of Chinese and express human leptin in E.coli.
Method:
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.
Conclusion:
Successful cloning and expression of human obesity gene in E.coli may facilitate further research of the mechanism of fat metabolism and adipocyte differentiation.
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