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。
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 with BamH Ⅰ and Pst Ⅰ 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.
出处
《蚌埠医学院学报》
CAS
2012年第1期11-13,共3页
Journal of Bengbu Medical College
基金
湖北省卫生厅科研基金资助项目(JX3B27)