[Assembly of full-length human resistin gene in vitro and construction of its eukaryotic expression vector].
To assemble the full-length of human resistin gene in vitro by using oligonucleotides and to construct its eukaryotic expression vector.



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.



The band of PCR products and gene sequencing showed the insert fragment in pSecTag2B vector was identical to that as designed.



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.
Qiao
Xiao-zhi
XZ
The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310003, China. qxz1971@hzcnc.com
Wang
Xian-feng
XF
Zhao
Dong-jiu
DJ
D001483:Base Sequence; D003001:Cloning, Molecular; D005813:Genes, Synthetic; D005822:Genetic Vectors; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D052243:Resistin / Q000235:genetics* / Q000378:metabolism
C499624:RETN protein, human; D011994:Recombinant Proteins; D052243:Resistin