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[Construction and expression of a prokaryotic vector of recombinant human adiponectin global domain]
1Department of Endocrinology, West China Hospital, Sichuan University, Chengdu, China.
Objective:
To construct and express the recombinant human adiponectin (gAd) global domain.
Methods:
gAd complementary DNA (cDNA) was obtained from human fat tissue by RT-PCR. The PCR product was cloned into the vector pMD18-T and the prokaryotic expression vector pET32a(+). The recombinant vector was identified by digestion with double restriction endonucleases SalI and EcoRI, PCR and sequence analysis. The recombinant plasmid containing gAd gene was transformed into E. coli BL21 (DE3), and the expression of the fusion protein His-gAd was induced by IPTG.
Results:
The gAd cDNA of 412 bp was obtained from the total RNA of the fat tissue and verified by sequence analysis.
Conclusion:
The recombinant plasmid could stably express the 34-kD fusion protein His-gAd in the engineered bacteria in the form of inclusion bodies.
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