Assembly of full-length human resistin gene in vitro and construction of its eukaryotic expression vector / 浙江大学学报·医学版

ABSTRACT

<p><b>OBJECTIVE</b>To assemble the full-length of human resistin gene in vitro by using oligonucleotides and to construct its eukaryotic expression vector.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>The band of PCR products and gene sequencing showed the insert fragment in pSecTag2B vector was identical to that as designed.</p><p><b>CONCLUSION</b>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.</p>