人巨细胞病毒pp65蛋白基因的克隆及表达  

Study on clone and express human cytomegalovirus pp65 protein.

作　　者：赵捷[1] 闻良珍[1] 

机构地区：[1]华中科技大学附属同济医院妇产科,武汉430020

出　　处：《中国优生与遗传杂志》2003年第1期40-42,共3页Chinese Journal of Birth Health & Heredity

摘　　要：目的　构建人巨细胞病毒pp6 5蛋白全长基因的原核表达载体 ,并对表达产物进行纯化。方法　PCR扩增pp6 5的全长基因 ,将其插入到原核表达载体 pRSET ,在工程菌BDPS中进行IPTG诱导表达 ,采用Westenblot进行鉴定并通过亲和层析对表达产物进行纯化。结果　成功构建了全长HCMVpp6 5的原核表达载体并诱导其高效表达 ,用镍离子柱亲和层析获取纯度达 90 %以上的表达蛋白。结论　我们获取的HCMVpp6 5原核表达蛋白纯度较高 ,可将其应用于免疫治疗和临床检测的进一步研究。Objective To construct human cytomegalovirus pp65 prokaryotic expressing vector and to express it, purify the product. Methods: The whole cDNA of HCMV pp65 was amplified by PCR and inserted into prokaryotic expressing vector pRSET by gene engineer technique; Recombinant plasmid was transformed into engineering bacteria BDPS and expressed by IPTG inducing; The product was identified use Western blot and purified by affinity chromphotography. Result: The pp65 prokaryotic expressing vector was successfully constructed and could express in engineering bacteria BDPS; Western blot analysis showed the product is just pp65 recombinant protein; High pure recombinant protein was acquired. Conclusion: The high pure recombinant protein is valuable for antibody preparation of human cytomegalovirus and can be used as a kind of potent specific antigen in human cytomegalovirus immunotherapy.

关 键 词：人巨细胞病毒 PP65蛋白 原核表达 

分 类 号：R346[医药卫生—基础医学]