摘要
目的克隆中国人的肥胖基因,并在大肠杆菌中高效表达人瘦素蛋白。方法用RT-PCR从培养的中国人脂肪细胞提取的总RNA中扩增出肥胖基因,与TA载体连接后进行核酸序列测定,并将该基因定向克隆至高效表达质粒pBV220,在大肠杆菌中表达。结果克隆出中国人肥胖基因,其cDNA序列与文献报道的一致,构建了重组质粒pBV220-OB,并成功地实现了人肥胖基因在大肠杆菌中的表达。结论人肥胖基因的克隆及其在大肠杆菌中的表达,为进一步研究该基因在肥胖症及其相关疾病中的作用,以及在脂肪代谢与脂肪细胞分化中的调控机制奠定基础。
Objective To clone the obesity gene of Chinese and express human leptin in E.coli. Method The obesity gene was amplified from the total RNA isolated from cultured human adipocytes of Chinese by reverse transcriptional PCR, inserted into TA-vector and cloned into the expression plasmid pBV220 after sequence identification. Results DNA sequencing confirmed that the isolated obesity gene was identical to the previously reported sequence. The recombinant plasmid pBV220-OB was constructed and leptin successfully expressed in E.coli. Conclusion Successful cloning and expression of human obesity gene in E.coli may facilitate further research of the mechanism of fat metabolism and adipocyte differentiation.
出处
《第一军医大学学报》
CSCD
北大核心
2005年第4期395-398,共4页
Journal of First Military Medical University
基金
国家自然科学基金(39880032)
广州市重点科技攻关项目(990448022)~~
关键词
肥胖症
肥胖基因
反转录PCR
基因表达
瘦素蛋白
obesity
obesity gene
reverse transcriptional polymerase chain reaction
gene expression
leptin
