跨膜蛋白66生物信息学分析及其突变体转基因小鼠构建  

Bioinformatic Analysis of TMEM66 Gene and Construction of TMEM66 Mutant Transgenic Mice

作　　者：李新云[1,2] 王趁芳[1] 任红艳[1] 赵书红[2] 杨述林[1] 崔文涛[1] 牟玉莲[1] 李奎[1] 

机构地区：[1]中国农业科学院北京畜牧兽医研究所家养动物遗传资源与种质创新重点开放实验室,北京100193 [2]华中农业大学动物遗传育种与繁殖教育部重点实验室,武汉430070

出　　处：《畜牧兽医学报》2010年第1期33-38,共6页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家863计划(2006AA10Z135;2008AA10Z143);国家973计划(2006CB102105);国家自然科学基金(30830080);中国农业科学院北京畜牧兽医研究所科技创新团队基金资助项目(ywf-td-1)

摘　　要：跨膜蛋白66(TMEM66)是与细胞凋亡以及癌症发生密切相关的重要功能基因,为了在个体水平研究其生物学功能,我们进行了TMEM66基因突变体(TMEM66V)转基因小鼠的构建。本研究通过原核注射法进行转基因操作,将获得的312个受精卵移植到13只代孕母鼠中,运用PCR、Southern blotting对出生的小鼠进行转基因鉴定,对于转基因阳性小鼠通过传代试验研究外源基因是否稳定整合,并通过反向PCR方法研究外源基因的整合方式。结果显示在出生的55只小鼠中有6只为转基因阳性,其中3只转基因小鼠可以稳定传代,表明这些小鼠中外源基因发生了稳定整合。反向PCR检测结果发现外源片段是以串联重复的方式整合到转基因小鼠的基因组中。本研究成功构建了TMEM66基因突变体转基因小鼠,为进一步研究TMEM66基因的功能奠定了基础。Transmembrane protein 66 play important roles in cell apoptosis and tumorigenesis.In order to investigate the function of TMEM66 gene,the transgenic mouse model containing the mutant TMEM66 gene was constructed in this study.The transgenic mice were generated by using microinjection method and 312 injected zygotes were implanted into 13 foster mothers.The positive transgenic mice were confirmed by PCR and Southern blotting.Also,the stability of the transgene was confirmed by passage and the way of integration of the transgene was studied using inverse PCR method.In this study,6 positive individuals among the 55 neonatal pups were found by Southern blotting.Three positive mice were found to be able to transfer the transgene to next generation.This result indicted that the transgene was integrated into host genome stably.Inverse PCR results indicated that the transgenic DNA fragment was integrated into the host genome in a serial multi-copy manner.Therefore,the transgenic mice model containing the mutant TMEM66 gene was constructed successfully.This study offered useful model for further investigation of the function of TMEM66 gene.

关 键 词：TMEM66基因 跨膜蛋白 转基因小鼠 

分 类 号：Q78[生物学—分子生物学]