[Assembly of full-length human resistin gene in vitro and construction of its eukaryotic expression vector]
Xiao-zhi Qiao1, Xian-feng Wang, Dong-jiu Zhao
1The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310003, China. qxz1971@hzcnc.com
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.
