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c-myb反义RNA重组载体构建及其包装细胞株的建立
引用本文:马会慧,姚集鲁,李刚,姚春斓,陈雪娟,杨绍基.c-myb反义RNA重组载体构建及其包装细胞株的建立[J].中山大学学报(医学科学版),2001,22(3):235-237.
作者姓名:马会慧  姚集鲁  李刚  姚春斓  陈雪娟  杨绍基
作者单位:中山医科大学附属第三医院传染科,
基金项目:广东省科委重点攻关项目(1977)
摘    要:【目的】构建反义c-myb重组逆转录病毒载体并建立其包装细胞株。【方法】采用RT-PCR方法,获取目的基因,通过TA克隆方法克隆入pUC19,再反向亚克隆入逆转录病毒载体pDOR中。采用DOTAP法将重组逆转录病毒载体转入包装细胞,以NIH3T3细胞为靶细胞测定产病毒滴度。【结果】序列测定结果与GenBank中序列一致。c-myb基因片段已定向克隆入pDOR。细胞转染表明,已形成稳定的包装细胞株PA317/pDOR-myb,产病毒滴度达5.2×104~9.5×104CFU/mL。【结论】成功构建了含有反义c-myb的重组逆转录病毒质粒,并建立其包装细胞株。

关 键 词:肝纤维化  c-myb  RNA,反义  克隆,分子  转染
文章编号:1000-257X(2001)03-0235-03
修稿时间:2000年8月23日

Construction of c-myb Antisense RNA Recombinant Retroviral Vector and Establishment of Its Packaging Cell Line PA317
MA Hui-hui,YAO Ji-lu,LI Gang,YAO Chun-Lan,CHEN Xue-juan,YANG Shao-ji.Construction of c-myb Antisense RNA Recombinant Retroviral Vector and Establishment of Its Packaging Cell Line PA317[J].Journal of Sun Yatsen University(Medical Sciences),2001,22(3):235-237.
Authors:MA Hui-hui  YAO Ji-lu  LI Gang  YAO Chun-Lan  CHEN Xue-juan  YANG Shao-ji
Abstract:【Objective】To construct the c-myb antisense RNA recombinant retroviral vector and its packaging cell line.【Methods】The segment of c-myb gene was cloned into pUC19 with TA cloning method after amplication by RT-PCR,and then was subcloned into retroviral vector pDOR.The recombinant retroviral vector named pDOR-myb was transfected into retroviral package cell line PA317 after selection with G418.【Results】Sequencing data indicated that the c-myb gene was exactly identical to the sequence in the GenBank.The segment of c-myb gene was inserted directionally into pDOR.Resistant colonies were obtained and the titers of pDOR-myb were 5.2×104~9.5×104 CFU/mL.【Conclusion】The recombinant retroviral vector containing c-myb gene is successfully constructed and its packaging cell line PA317/pDOR-myb was established.
Keywords:liver fibrosis  c-myb  RNA  antisense  cloning  molecular  transfection
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