激光生物学报2016，Vol.25Issue(6)：546-552,7.DOI:10.3969/j.issn.1007-7146.2016.06.010
斑马鱼心脏发育候选基因Klhl31的TALEN打靶有效性分析
Targeting Effectiveness Analysis of Heart Developmental Candidate Gene Klhl31 with Gene Knockout Method TALEN
摘要
Abstract
TALEN (transcription activator-like effector nuclease) targeting technology is a method of gene knockout orientation,which can solve the problem that is difficult for in vivo gene knockout in zebra fish at present.Klhl31 is one of human heart developmental candidate genes identified in our laboratory.So far there is no Klhl31 gene knockout strains in zebra fish at home and abroad.In order to identify the biological function of Klhl31 gene in zebra fish,we used TALEN technique to build Klhl3 1 gene knockout strains in zebra fish.Digestion analysis showed that mutational DNA band occurred at the cleavage site in those injected embryos with TALEN expression plasmids,suggesting that the restriction sites were destroyed,which demonstrated that TALENs technology was effective in this experiment.In the selection of F0 generation of mutants,the positive results were confirmed by enzyme digestion.These results suggested that the application of TALEN techniques to knockout Klhl31 gene was successful,and the chimerical Klhl31 knockout zebrafish was obtained.关键词
Klhl31/TALEN/基因突变/酶切位点/基因敲除Key words
Klhl31/TALEN/gene mutations/restriction sites/gene knockout分类
生物科学引用本文复制引用
蔡湾湾,陈发,万永奇,吴秀山,邓云,袁婺洲..斑马鱼心脏发育候选基因Klhl31的TALEN打靶有效性分析[J].激光生物学报,2016,25(6):546-552,7.基金项目
国家自然科学基金(81170229,81370451,31472060,81170088) （81170229,81370451,31472060,81170088）
湖南省研究生科研创新基金(CX2011B195) （CX2011B195）
湖南省生物发育工程及新产品研发协同创新中心(2013-448-6) （2013-448-6）
