摘要
目的 克隆HER 2 /neu胞外配体结合区基因 ,并在大肠杆菌中获得高效表达。方法 用PCR技术扩增HER 2 /neu胞外配体结合区的cDNA片段 ,并将该片段插入pET 2 8a(+)原核表达载体中 ,实现插入基因的融合表达 ;用SDS PAGE和蛋白免疫印迹法分别测定表达产物的相对分子质量及特异性。结果 构建的重组质粒在大肠杆菌中获得高效稳定的表达 ,表达产物的相对分子质量与预期值一致 ,且所表达蛋白可被抗HER 2 /neu的特异性抗体识别。结论 获得了HER 2 /neu胞外配体结合区基因在原核系统中的表达 ,为HER 2
Objective To clone and express the gene of HER 2/neu ext racellular domain.Methods The cDNA fragment of HER 2/neu extra cellular domain was amplified by PCR, then cloned into the prokaryotic phagemid pET 28a(+) vector, and expressed as a fusion protein in E.coli . SDS PAGE a nd Western blotting were used to analyze the molecular weight and specificity o f the expressed protein. Results The recombinant protein was express ed highly and stably in E.coli , and the molecular weight of the expression p rodu ct was identical to the expected value. In addition, the protein expressed could react with the specific antibody against protooncoprotein HER 2/neu. Conclusion A plasmid containing HER 2/neu extracellular domain gene was constructed. The expression of the HER 2/neu extracellular protein was obtained in E.coli . It will be helpful to the study of the vaccine for the tumor ove rexpressing HER 2/neu oncoprotein.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2003年第3期183-185,189,共4页
Immunological Journal
基金
国家自然科学基金资助项目 (30 0 70 85 8)
