Site-directed mutagenesis of long QT syndrome KCNQ1 gene in vitro

作     者：Wei LI Junguo YANG Rong DU Li TIAN Bin WANG Qiumei XU Qinmei KE Qing WANG 

作者机构：Geriatric DepartmentUnion HospitalTongji Medical CollegeHuazhong Science and Technology UniversityWuhan 430022China Institute of CardiologyUnion HospitalTongji Medical CollegeHuazhong Science and Technology UniversityWuhan 430022China Center of Cardiovascular Geneticsthe Cleveland Clinic FoundationClevelandOhio 44195USA 

出 版 物：《Frontiers of Medicine》 (医学前沿（英文版）)

年 卷 期：2008年第2卷第1期

页      面：100-104页

核心收录：

学科分类：1002[医学-临床医学] 100214[医学-肿瘤学] 10[医学] 

基　　金：study of LQTS mutation function is the next step.Inthis cxperimcnt,wc construced the eukaryortic expression vector of pIRES2-EGFP-KCNQ1(G983A) expressed it in HEK293 cells successfully which sheds light on the functional studies of the KCNQ1 gene 

主　　题：long QT syndrome KCNQ1 polymerase chain reaction site-directed mutagenesis 

摘      要：To construct a polymerase chain reaction(PCR)site-directed mutagenesis of the long QT syndrome KCNQ1 gene in vitro,two sets of primers were designed according to the sequence of KCNQ1 cDNA and a mismatch was introduced into *** was performed in a two-step *** amplified fragments from the third PCR which contained the mutation site were sub-cloned into the T-vector ***,the fragments containing the mutation site was obtained from pCR2.1 using restriction enzymes digestion and inserted into the same restriction site of *** sequencing analysis shows that the mutation site was *** from A to G in site 983 of KCNQ1 cDNA was *** the Effectene transfection reagent,pIRES2-EGFP-KCNQ1(G983A)was transfected into HEK cells *** results may shed light on further functional study of KCNQ1 gene.