HCV不同编码区重组基因的蛋白表达丰度分析  

The abundance assay of protein expression from different recombinant gene fragments of hepatitis C virus

作　　者：乔伟振[1] 孟继鸿[2] 董晨[2] 谢平[1] 杨燕[1] 李小雯[1] 

机构地区：[1]南京医科大学附属无锡市人民医院中心实验室,江苏无锡214023 [2]东南大学病原生物学与免疫学系,江苏南京210009

出　　处：《南京医科大学学报（自然科学版）》2008年第6期722-725,766,共5页Journal of Nanjing Medical University(Natural Sciences)

基　　金：南京医科大学科技发展基金(NY0583);南京医科大学科技发展基金重点项目(07NMUZ034)

摘　　要：目的:探讨丙型肝炎病毒(HCV)不同基因片段重组载体在pET大肠杆菌系统中的蛋白表达情况。方法:分别将构建好的含有HCVC、NS3、NS5片段基因的pET重组质粒转化至大肠杆菌BL21(DE3)中,通过卡那霉素抗性筛选转化子。经过IPTG诱导表达相应的重组蛋白,通过Ni-NTA凝胶亲和层析纯化各目标蛋白,并分别计算各重组蛋白的浓度。结果:分别诱导表达含有HCVC、NS3、NS5片段基因的重组蛋白,各蛋白的浓度分别为:3.557(1型NS3)、4.238(6型NS3)、2.087(NS5)、1.490g/L(C)。结论:在相同条件下,诱导表达含有HCV不同基因片段的重组载体,所得到的蛋白丰度存在差异。Objective：To explore the abundance of recombinant proteins expressed by the pET/E.coli system from different HCV gene regions. Methods：The four recombinant plasmids（pET-C／pET-NS5／pET-NS3-1／pET-NS3-6） were all transformed into the E.coli BL21（DE3） and screened by Kanamycin（50 μg/ml）. After induced by isopropyl-β-D-thiogalactopyranoside（IPTG）,the four expression products of fusion proteins were detected by 15% sodium dodecyl sulfate（SDS）-polyacrylamide gel electrophoresis（PAGE）,and were purified by Ni-NTA Resin chromalography. The absorbances of the four recombinant proteins were measured by spectro photometer,and then the every protein concentration was calculated,respectively. Results：The four recombinant protein concentrations were 3.557（NS3 type 1）,4.238（NS3 type 6）,2.087（NS5） and 1.490 g/L（C）,respectively. Conclusion：The result indicated that the abundance of four recombinant proteins was apparently different,derived by pET/E.coli system with different HCV gene fragments in the same condition.

关 键 词：丙型肝炎病毒 pET系统 大肠杆菌 蛋白表达 

分 类 号：R392.11[医药卫生—免疫学]