腺相关病毒和人γ-干扰素基因重组体转染淋巴细胞的研究  

Transfection and expression of the recombinant adeno-associated virus carrying human interferon-γ in human lymphocyte

作　　者：朱海红[1] 陈智[1] 章明太[1] 姚航平[1] 丁列明 

机构地区：[1]浙江大学医学院附属一院传染病研究所,杭州310003 [2]美国阿肯萨斯大学病理系

出　　处：《中华传染病杂志》2000年第2期77-79,共3页Chinese Journal of Infectious Diseases

基　　金：国家自然科学基金!( 3 95 70 65 3 )

摘　　要：目的　观察腺相关病毒和人γ 干扰素的重组体 pWIG转染淋巴细胞及在细胞内表达情况。方法　将 pWIG以DNA 磷酸钙共沉淀法转染人淋巴细胞株H9,再分别用聚合酶链反应 (PCR)、逆转录 PCR(RT PCR)和酶联免疫吸附试验 (ELISA)等方法在DNA、RNA和蛋白质水平研究该重组体导入细胞和在细胞内表达的情况。结果　转染 pWIG的H9细胞的DNA和RNA经PCR和RT PCR检测均有 5 0 4bp的DNA条带 ;用ELISA可测出γ 干扰素的表达量为 17.2 pg/ 3× 10 6细胞。结论　重组体 pWIG用DNA 磷酸钙共沉淀法导入细胞后 ,转录HuIFN γmRNA ,并翻译人γ 干扰素蛋白。Objective In order to study the transfection and expression of the recombinant pWIG of Adeno associated virus and Human interferon γ(HuIFN γ) in human lymphocyte.Methods After transfecting pWIG into human lymphocyte via calcium phosphate, DNA, RNA and protein extracted from transfected cells were detected by polymerase chain reaction(PCR), reverse transcription PCR(RT PCR) and ELISA. Results The 504 bp bands were detected from PCR and RT PCR products of the DNA and RNA from the cells transfected, and HuIFN γ protein were detected to be 17.2 pg/3×10 6 cells by ELISA.Conclusion The recombinant pWIG could produce HuIFN γ mRNA after the transfection via calcium phosphate, and translate corresponding protein further. This lays ground for the further research of gene therapy.

关 键 词：十扰素γ 腺 相关病毒 基因治疗 载体系统 pWIG 

分 类 号：R394.8[医药卫生—医学遗传学]