论文部分内容阅读
目的 在昆虫细胞中表达人乳头瘤病毒 16型 (HPV16 )结构蛋白 ,为HPV16型预防性基因工程亚单位疫苗的研究以及诊断试剂的研制奠定基础。方法 将目的基因克隆至杆状病毒转移载体 ,该重组质粒DNA与线性化的杆状病毒DNA共转染昆虫细胞Sf 9,通过同源重组获得重组杆状病毒 ,用SDS PAGE凝胶电泳和Westernblot法检测重组子在昆虫细胞中表达的目的蛋白。结果 获得了稳定高效表达HPV16结构蛋白的重组杆状病毒 ,L1和L2蛋白的相对分子质量分别为 5 70 0 0和970 0 0 ,L1蛋白的表达产量约占昆虫细胞总体蛋白的 2 5 %~ 30 %。结论 人乳头瘤病毒 16型结构蛋白可在昆虫细胞内高效表达
Objective To express human papillomavirus 16 (HPV16) structural protein in insect cells and lay a foundation for the research of HPV16 preventive genetic engineering subunit vaccine and the development of diagnostic reagents. Methods The target gene was cloned into the baculovirus transfer vector. The recombinant plasmid DNA and the linearized baculovirus DNA were co-transfected into insect cells Sf 9, and the recombinant baculovirus was obtained by homologous recombination. SDS PAGE gel electrophoresis and Western blot Method to detect the recombinant protein expressed in insect cells target protein. Results The recombinant baculovirus expressing HPV16 structural protein was successfully and efficiently expressed. The relative molecular weights of L1 and L2 proteins were respectively 5 70 0 0 and 9 70 0 0. The expression of L1 protein accounted for 25% 30%. Conclusion Human papillomavirus type 16 structural protein can be efficiently expressed in insect cells