| DNMT1靶向RNA干扰重组体的构建及序列分析 |
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| 引用本文： | 陈道荣,王丕龙,黄爱龙.DNMT1靶向RNA干扰重组体的构建及序列分析[J].重庆医科大学学报,2004,29(3):269-272. |
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| 作者姓名： | 陈道荣 王丕龙 黄爱龙 |
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| 作者单位： | 重庆医科大学临床学院消化内科,重庆,400016;重庆医科大学病毒性肝炎研究所,重庆,400010 |
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| 摘 要： | 目的:本研究利用RNA干扰(RNA interfering,RNAi)技术,以DNMT1(DNAmethyltransferase 1)为靶基因,设计构建重组体,并进行序列分析,为下一步探索肿瘤基因治疗的新途径打好基础.方法:设计有小发夹结构的两条DNA序列,经退火成互补双链,再克隆至载体pTZU6 1中构建重组体,转化JM109菌株,提取质粒行酶切鉴定后,进行测序分析.结果:将合成的DNA序列退火后克隆到载体上,经测序鉴定确实为所需序列.结论:DNMT1靶向RNA干扰重组体的成功构建和序列分析,可进一步利用克隆所得的小RNA序列干扰DNMT1的mRNA转录,从而达到治疗肿瘤的目的.
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| 关 键 词： | DNMT1 RNA干扰 重组体 序列分析 |
| 文章编号： | 0253-3626(2004)03-0269-04 |
| 修稿时间： | 2003年9月1日 |
Cloning and sequence analyzing of the recombinant plasmid affecting gene DNMT1 translation by RNA interfering |
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| CHEN Daorong,et al.Cloning and sequence analyzing of the recombinant plasmid affecting gene DNMT1 translation by RNA interfering[J].Journal of Chongqing Medical University,2004,29(3):269-272. |
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| Authors: | CHEN Daorong |
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| Abstract: | Objective:To clone the recombinant plasmid affecting gene DNMT1 translation by RNA interfering and analyze the nucleic acid sequence of the recombinant for further searching new gene therapy method of tumor.Methods:Two DNA sequences containing small hairpin structure were designed and synthesized.The complement form was obtained by annealing and inserted into vector pTZU6 1,and the recombinant plasmid was transformed into Jm109 strain.Finally the plasmid identified by restriction enzyme was used for sequence analysis.Results:The recombinant was cloned and the aim sequence was obtained.Conclusion:Successful cloning of the recombinant helps to search new gene therapy method of tumor. |
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| Keywords: | DNMT1 RNA interfering Cloning Sequence analysing |
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