Assembly of full-length human resistin gene in vitro and construction of its eukaryotic expression vector.
- Author:
Xiao-zhi QIAO
1
;
Xian-feng WANG
;
Dong-jiu ZHAO
Author Information
- Publication Type:Journal Article
- MeSH: Base Sequence; Cloning, Molecular; Genes, Synthetic; Genetic Vectors; Humans; Molecular Sequence Data; Polymerase Chain Reaction; methods; Recombinant Proteins; genetics; metabolism; Resistin; genetics; metabolism
- From: Journal of Zhejiang University. Medical sciences 2007;36(6):588-591
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo assemble the full-length of human resistin gene in vitro by using oligonucleotides and to construct its eukaryotic expression vector.
METHODSAccording 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.
RESULTSThe band of PCR products and gene sequencing showed the insert fragment in pSecTag2B vector was identical to that as designed.
CONCLUSIONThe full-length of human resistin coding sequence was successfully assembled and amplified by touch down PCR, and a resistin-expressing eukaryotic vector was constructed.
