关岭牛pEGFP-N_3-UCP3重组真核表达载体的构建

Construction of Recombinant Eukaryotic Expression Vector of pEGFP-N_3-UCP3 in Guanling Cattle

旨在构建不含CMV启动子的p EGFP-N3-UCP3重组真核表达载体。利用PCR技术克隆UCP3基因启动子,扩增不含CMV区的p EGFP-N3载体片段,将UCP3启动子与该载体经连接、转化后,挑取阳性克隆进行PCR鉴定及测序鉴定。结果表明,成功地构建了重组载体p EGFP-N3-UCP3,为UCP3基因转录调控机制的研究奠定了基础。

This study was carried out to construct a recombinant eukaryotic expression vector of pEGFP-N3-UCP3 without CMV promoter.UCP3 gene promoter and pEGFP-N3 vector fragment without CMV region was cloned and amplified by PCR assay.Subsequently,the obtained UCP3 gene promoter and the amplified vector were connected and transformed into competent cell DH5αfrom Escherichia coli.The positive clone was selected and identified by PCR assay and sequencing.The recombinant eukaryotic expression vector of pEGFP-N3-UCP3 was successfully constructed,which laid a foundation for uncovering the mechanism underlining the transcriptional regulation of UCP3 gene.