扎伊尔埃博拉病毒糖蛋白基因的克隆和表达

Cloning and expression of Zaire Ebola virus glycoprotein in prokaryotic and eukaryotic cells

埃博拉病毒属丝状病毒科，能引发动物和人出血热症状，人感染后病死率高达90％以上，目前还没有有效预防和治疗的药物和疫苗。近年来，这种烈性传染病病毒传入我国的可能性不断加大，给我国公共卫生应急体系带来新的挑战。本研究针对埃博拉病毒的最主要结构蛋白———糖蛋白（GP），构建了重组原核表达载体pET28a（＋）‐GP1（33～313 aa）、pET28a（＋）‐GP1（190～313 aa）、pET28a（＋）‐GP2（502～632 aa）、pET28a（＋）‐sGP ，以及重组真核表达载体pcDNA3．1（＋）‐edited GP、pcDNA3．1（＋）‐GP1、pcDNA3．1（＋）‐GP。结果表明，GP1（33～313 aa）、GP1（190～313 aa）和sGP能在大肠埃希菌BL21（DE3）中以包涵体的形式表达，GP、GP1和GP2能在HEK293T细胞中表达，但均不能在BHK21细胞中表达。本研究为进一步探索埃博拉病毒GP的结构和功能及GP抗体制备奠定了基础。

Ebola virus belongs to filoviridae .Zaire Ebola virus has a high mortality rate up to 90% .It is transmitted through blood , secretions , organs or other body fluids of infected people or animals . Unfortunately ,there are still no approved drugs and vaccines for Ebola virus infection .Considering the increasing commercial and personal communication with countries in Africa ,the possibility of Ebola virus invading into China by infected animals and human highlights the necessity for the development of effective vaccines .In this study ,a series of glycoproteins （GPs） were expressed by using pET28a（＋ ） and pcDNA3 .1 （＋ ） in prokaryotic and eukaryotic cells ,respectively .The results showed that GP1（33‐313 aa） ,GP1（190‐313 aa） and nonstructural secretory glycoprotein （sGP） were expressed in Escherichia coli BL21 （DE3）in inclusion bodies ,while no GP2 （502‐632 aa） expression could be detected under the same condition .The edited GP ,GP1 and GP2 were expressed in HEK293T cells ,but not in BHK21 cells .The data provide some clues for the development of antiviral serum and effective drugs for the treatment of Ebola virus infection .