摘要
利用E.coli原核表达系统克隆、重组表达截短的人血管紧张素转化酶2(ACE 2)。通过细胞分子生物技术获取截短的人ACE2的基因,并将其克隆到p ET28a载体上,且在大肠杆菌BL21(DE3)中重组表达该截短的人ACE 2蛋白。获取的截短的人ACE2基因,通过Nco I/Xho I双酶切后,将其克隆到经过同样Nco I/Xho I双酶切的p ET28a表达载体上,得到p ET28a-h ACE2重组质粒。然后将p ET28a-h ACE2重组质粒转化到大肠杆菌BL21(DE3)中,在37℃1 mmol/L IPTG诱导下重组表达了截短的人ACE2,通过免疫学方法初步鉴定正确表达了该ACE2。在原核表达系统中,重组表达截短的人ACE2;为研究ACE2的结构与功能的关系奠定了一定研究基础。
Truncated human angiotensin-converting enzyme 2( ACE2) protein is recombinantly expressed in E. coli prokaryotic expression system. A truncated human ACE2 gene of interest was obtained by the method of molecular cell biology. The gene was cloned into the vector p ET28 a. The truncated human ACE2 protein was recombinantly expressed in E. coli BL21( DE3) and characterized with SDS-PAGE and Western blot. The truncated human ACE2 gene was got in this study. After digestion with Nco I / Xho I double enzymes,the gene was successfully cloned into the vector p ET28 a which was also digested with the same Nco I / Xho I. A recombinant vector p ET28a-h ACE2 was formed and transformed into E. coli BL21( DE3). The recombinant h ACE2 was expressed after 4 hours with induction of 1 mmol / L IPTG at 37 ℃ and identified using Western blot. In prokaryotic expression system,the truncated human ACE2 was successfully expressed. It will provide research basis for the relationship between structure and function of ACE2.
出处
《药物生物技术》
CAS
2014年第5期398-401,共4页
Pharmaceutical Biotechnology
关键词
血管紧张素转化酶2
大肠杆菌
克隆
蛋白表达
重组质粒
Angiotensin-converting enzyme 2,E.coli,Clone,Protein expression,Recombinant plasmid
