邢念增 张智清 郭应禄 俞莉章 刘漓波 陈庆华 朱丽华 钭理强 100034 北京医科大学第一医院泌尿外科、北京医科大学泌尿外科研究所(邢念增、郭应禄、俞莉章、刘漓波、钭理强)，中国预防医学科学院病毒学研究所病毒基因工程国家重点实验室(张智清、陈庆华)，北京医科大学第一医院检验科(朱丽华) 中华泌尿外科杂志 1998 7 19 7


    关键词：逆转录病毒载体；基因；肾肿瘤；癌 期刊 zhmnwkzz 0 论 著 fur -->


    

摘要 将人B7基因亚克隆到逆转录病毒载体pLXSN中，构建了人B7逆转录病毒载体

    pLXSNB7，经Lipofectin介导转染包装细胞，并通过G418筛选获得较高病毒滴度的细胞株

    PA317/pLXSNB7(3×10⁵ ～1×10⁶ cfu/ml)。用此逆转录病毒基因转移体系将B7基因转移

    到肾癌GRC-1细胞中，经RT-PCR、间接免疫荧光及LSAB细胞免疫组化等检测方法检测到

    转基因细胞中B7mRNA及B7分子的表达。介导人B7基因的逆转录病毒基因转移体系的建立，

    为人肾癌转基因瘤苗的应用研究提供了实验基础。

Construction ofretroviral delivery system mediating human B7 gene and B7 expression in renal cellcarcinoma cells Xing Nianzeng, Zhang Zhiqing, Guo Yinglu, et al. Department ofUrology, the First Affi-liated Hospital, Beijing Medical University. Institute of Urology,Beijing Medical University, Beijing 100034

    Abstract The expressing plasmid pLXSNB7was constructed by subcloning human B7 gene into the retroviral vector pLXSN, pLXSNB7 wasthen transfected into package cell line PA317 using Lipofectin and finally PA317/pLXSNB7cell strains were obtained yielding 3 ×10⁵ ～ 1 ×10⁶ cfu/ml virus titer after G418 selection. B7mRNA of the transfected tumor cells was detected by RT-PCR. Indirect immunofluorescentmicroscopy and immunohistochemistry showed the expression of B7 on the surface of theinfected tumor cells. The experiment laid the foundation to develop B7 gene-gransducedtumor vaccine of renal cell carcinoma.

    Key words Retroviral vector Genes Kidney neoplasms Carcinoma

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