文章摘要
胡晓山,余章斌,李萌萌,朱金改,朱 春,韩树萍.miR-19b转基因小鼠的构建及表型分析[J].南京医科大学学报,2013,(5):579~585
miR-19b转基因小鼠的构建及表型分析
Construction and phenotype analysis of miR-19b transgenic mouse
投稿时间:2012-12-07  
DOI:10.7655/NYDXBNS20130504
中文关键词: 转基因小鼠  心脏肥大  miR-19b  心血管疾病
英文关键词: transgenic mice  cardiac hypertrophy  miR-19b  cardiovascular disease
基金项目:国家自然科学基金(81070500);江苏省医学重点人才基金项目(RC2011021)
作者单位
胡晓山 南京医科大学附属南京市妇幼保健院儿科,江苏 南京 210004 
余章斌 南京医科大学附属南京市妇幼保健院儿科,江苏 南京 210004 
李萌萌 南京医科大学附属南京市妇幼保健院儿科,江苏 南京 210004 
朱金改 南京医科大学附属南京市妇幼保健院儿科,江苏 南京 210004 
朱 春 南京医科大学附属南京市妇幼保健院儿科,江苏 南京 210004 
韩树萍 南京医科大学附属南京市妇幼保健院儿科,江苏 南京 210004 
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中文摘要:
      目的:构建心肌细胞特异性过表达miR-19b的转基因小鼠,并对其进行表型分析。方法:先确定载体插入的酶切位点,根据CloneEZ○R PCR Cloning Kit重组原理设计相应引物,以小鼠基因组为模板克隆出带有miR-19b前体序列(pre-miR-19b)的目的片段,利用重组酶连接到带有alpha-MHC 启动子的载体上。将所得载体线性化后,采用显微注射方法注射到小鼠胚胎干细胞内。再将所得的重组胚胎干细胞移植入假孕的C57BL/6J雌鼠子宫内,在所得的子代,即Founder小鼠中采用特异的引物鉴定出转基因小鼠,并选出相应的品系,进行表型分析。结果:成功构建miR-19b转基因小鼠,且miR-19b转基因小鼠舒张期室间隔厚度?舒张期左室游离壁厚度以及射血分数均有升高。结论:心肌特异性过表达miR-19b可以导致心脏肥大。
英文摘要:
      Objective:To construct heart-special miR-19b overexpression transgenic mouse and analyze their phenotype. Methods:Firstly,we found out the suitable restriction enzyme cutting site on the plasmid which carrying alpha-MHC promoter,and design the primers according recombination principle provided by CloneEZR PCR Cloning Kit. We inserted the pre-miR-19b sequence cloned from mice chromosome into the plasmid using recombinase. Secondly,after linearization of recombinant plasmid,they were injected into the embryonic stem(ES) cells. We transplanted the recombinant ES cells into the uterus of the pseudocyesis female C57BL/6J mice. At last,we selected out the lines of transgenic mouse and analyzed the phenotype of TG mouse. Results:We successfully constructed miR-19b transgenic mouse. The scores of interventricular septal thickness,left ventricle posterior wall and ejection fraction of miR-19b transgenic mouse were increased. Conclusion:The overexpression of heart-special miR-19b may lead to cardiac hypertrophy.
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