SDF-1α基因真核表达载体的构建与鉴定

Construction and Identification of Eukaryotic Expression Vector of SDF-1α Gene

目的构建SDF-1α真核表达载体,为研究SDF-1α基因功能提供工具。方法从大鼠平滑肌细胞中提取总RNA,采用RT-PCR技术扩增SDF-1α的基因编码区序列,将序列定向克隆至真核表达载体pcDNA3.1上,并用PCR、酶切法和DNA测序鉴定。结果PCR和限制性内切酶分析鉴定,表明获得重组质粒pcDNA3.1/SDF-1α。通过对SDF-1α基因编码区cDNA序列测序证实其与GenBank中的已知序列一致。结论成功构建了SDF-1α真核表达载体,为SDF-1α基因功能研究奠定基础。

Objective To construct an eukaryotic expression vector of SDF-la gene and to provide a tool for studying its function. Methods The cDNA of SDF-1α was amplified by RT-PCR and PCR. After purification, the gene was cloned into eukaryotie expression vector pcDNA3.1. The constructed recombinant plasmid was identified by PCR confirmation, enzyme digestion and DNA sequencing. Results The constructed recombinant plasmid pcDNA3.1/SDF-1α was obtained, positive clones were screened and identified by PCR and digestion with restriction enzyme. The sequence of SDF-1α was in accordance with the expected one. Conclusion The eukaryotic expression vector of SDF-1α gene has been successfully constructed, which may provide a basis for further researches.