Cloning and sequence analyzing of the recombinant plasmid affecting gene DNMT1 translation by RNA interfering

CHEN Daorong; et al

doi:10.13406/j.cnki.cyxb.2004.03.004

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Cloning and sequence analyzing of the recombinant plasmid affecting gene DNMT1 translation by RNA interfering

更新时间：2024-01-17

- Cloning and sequence analyzing of the recombinant plasmid affecting gene DNMT1 translation by RNA interfering
- Journal of Chongqing Medical University Issue 3, Pages: 269-272(2004)
- 作者机构：
  
  1. 重庆医科大学临床学院消化内科
  2. 重庆医科大学病毒性肝炎研究所,重庆,400016
  3. ,重庆,400010
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.2004.03.004
  CLC： R73-36
- Published：2004，
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[1]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(03):269-272.

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 (3):269-272(2004)
[1]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(03):269-272. DOI： 10.13406/j.cnki.cyxb.2004.03.004.

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 (3):269-272(2004) DOI： 10.13406/j.cnki.cyxb.2004.03.004.

摘要

目的 :本研究利用RNA干扰（RNAinterfering

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JM10 9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

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.

关键词

DNMT1RNA干扰重组体序列分析

Keywords

DNMT1RNA interferingCloningSequence analysing

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