Assembly of full-length human resistin gene in vitro and construction of its eukaryotic expression vector

Objective: To assemble the full-length of human resistin gene in vitro by using oligonucleotides and to construct its eukaryotic expression vector.Methods: According to the gene sequence of resistin(GenBank:AF323081),10 oligonucleotides were designed and synthesized,followed by a touch down PCR to assemble the full-length gene.The PCR products were cloned into pSecTag2B vector and confirmed by sequencing.Results: The band of PCR products and gene sequencing showed the insert fragment in pSecTag2B vector was identical to that as designed.Conclusion: The full-length of human resistin coding sequence was successfully assembled and amplified by touch down PCR,and a resistin-expressing eukaryotic vector was constructed.