人脂联素重组克隆载体构建

Reconstruction of Eukaryotic Expression Vector Containing Human Adiponectin Gene

目的:构建含有人脂联素(adiponectin,ad)基因的重组克隆载体pEGFP-N1-AD,为进一步研究AD与脂质代谢及代谢综合征关系奠定基础。方法:根据Gene-Bank中已经公布的AD设计引物,从含有目的基因的质粒克隆模板中,利用PCR方法钓取目的基因。将AD基因克隆到真核表达载体pEGFP-N1中。酶切、PCR、及测序鉴定。结果:PCR、酶切及测序鉴定证实目的基因正确克隆至真核表达载体pEGFP-N1中,测序结果与Gene-Bank报道一致。结论:成功构建了AD重组克隆真核表达载体。

Objective： To construct the vector （pEGFP-N1-AD） containing human adiponectin （AD）gene so as to investigate the effects of adiponectin in the lipid metabolism and the syndrome of metabolism. Methods： AD gene was amplified from the plasma by polymerase chain reaction （PCR） technique whose primer was designed according to the announced sequences ofAD gene in gene bank, then cut by the endonucleases and directionally cloned into the pEGFP-N1 vector and analyzed by endonuclease cutting,PCR and gene sequencing analysis. Results： Through PCR, endonuclease cutting and gene sequencing,the target gene was verified to be correctly cloned to vector pEGFP-N1. Conclusion： Recombinant eukaryotic expression vector containing human adiponectin protein was successfully constructed.