%0 Journal Article
%T PCR Site-Directed Mutagenesis of Long QT Syndrome KCNQ1 Gene in vitro<br>应用PCR技术对先天性长QT综合征KCNQ1 基因进行定点突变的研究PCR Site-Directed Mutagenesis of Long QT Syndrome KCNQ1 Gene in vitro
%A LI Wei
%A YANG Jun-Guo
%A REN Fa-Xin
%A KANG Cai-Lian
%A ZHANG Shou-Yan
%A <br>李伟
%A 杨钧国
%A 任法鑫
%A 康彩练
%A 张守焰
%J 遗传
%D 2004
%I 
%X To study PCR site-directed mutagenesis of long QT syndrome KCNQ1 gene in vitro.The site-directed mutagenesis of LQTS gene KCNQ1 was made by PCR.Two sets of primers were designed according to the sequence of KCNQ1 cDNA,and mismatch was introduced into primers.Mutagenesis was performed in a three-step PCR.The amplified fragments from the third PCR which contained the mutation site were subcloned into the T-vecor PCR 2.1.Then the fragments containing the mutation site was obtained from PCR2.1 with restriction enzyme digestion and was inserted into the same restriction site of pIRES_2-EGFP-KCNQ1.With Effectene Transfection Reagent,pIRES_2-EGFP-KCNQ1 was transfected into HEK293 cell.The sequencing analysis showed that the mutation site was correct.Mutation from T to C in 934 site of KCNQ1 cDNA was found.Under the fluorescence microscope,the green fluorescence was spread in the transfected HEK293 cell,meaning the pIRES_2-EGFP-KCNQ1 containing the mutation site was expressed correctly.
%K KCNQ1
%K PCR<br>长QT综合征
%K 定点突变
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=3E23F50E071DF18E21B2F5AEE4F1FA5E&aid=F8196B4A91A125D0&yid=D0E58B75BFD8E51C&vid=96C778EE049EE47D&iid=94C357A881DFC066&sid=4081E94A71AB3C30&eid=2EAE52BA5B1222A9&journal_id=0253-9772&journal_name=遗传&referenced_num=0&reference_num=7