DNMT1、DNMT3b siRNA重组质粒的构建及鉴定

Construction and identification of siRNA recombinant plasmid of DNMTl and DNMT3b genes

摘要: 目的：构建DNA甲基转移酶（DNMT1）、DNMT3b微干扰RNA（siRNA）重组质粒，并鉴定其正确性。方法：构建DNMT1siRNA重组质粒和DNMT3b siRNA重组质粒各4种，分别用BamHⅠ和PstⅠ酶切、测序、鉴定。结果：用BamHⅠ和PstⅠ进行双酶切后所有质粒均为阳性重组载体，每一种载体选择两个克隆进行测序鉴定，测序结果与插入的寡核苷酸的序列完全相符。结论：DNMT1 siRNA重组质粒和DNMT3b siRNA重组质粒正确，可用于后续实验。

Abstract: Objective:To construct and identificate the siRNA recombinant plasmid of DNMT1 and DNMT3b genes.Methods:Four types of siRNA recombinant plasmid of DNMT1 or DNMT3b gene were constructed,and identificated.Results:After digested withBamH Ⅰ and pstⅠ enzymes,all plasmids showed positive for recombinant vector plasmids.Two clones of each type of vector plasimid were selected for,sequencing identification.The results of sequencing identification were consistent with the inserted oligonucleotides.Conclusions:The siRNA recombinant plasmids of DNMT1 and DNMT3b genes were constructed corectly and could be used for further study.