摘要
目的 为了获得大量重组HCVE2蛋白 ,以研究E2抗体潜在的保护作用。方法 利用PCR方法从HCV基因组序列中扩增出 931bp的E2基因片段 ,经EcoRI和Sall双酶切后连接到pET 2 8a表达载体上 ,转化大肠杆菌BL2 1(DE3)菌株 ,得到重组质粒PET -E2 ,工程菌经IPTG诱导培养 ,明显表达出HCVE2蛋白 ,表达产物经固定化金属配体亲和层析纯化 ,用ELISA方法检测生物学活性。结果 表达产物主要以包涵体形式存在 ,表达量达菌体蛋白的 18%以上 ,目的蛋白具有良好的反应原性。
Objective To obtain large amounts of recombinant HCV E2 protein and study on the potential protection of it.Methods A 931bp of E2 gene fragment was amplified from HCV genome and transformed to E.coli BL21(DE3). A recombinant plasmid pET E2 was constructed in this way and expressed under the induction of IPTG. The expressed HCV E2 protein was purified by immobilized metal ion affinity chromatography(IMAC), and the biological activity of it was detected by ELISA. Results More than 18% of somatic protein was expressed. The expressed product showed good reactogenicity,and most of them were inclusion bodies.Conclusion The cloning and expression of HCV E2 gene laid a foundation of further study on HCV E2 protein and DNA vaccine.
出处
《中国生物制品学杂志》
CAS
CSCD
2000年第3期144-146,150,共4页
Chinese Journal of Biologicals
