几丁质酶结构域内含蛋白1(Chid1)基因剔除小鼠的建立  

Generation of Chitinase Domain Containing 1 (Chid1) Knockout Mice

作　　者：奚骏 吴友兵 孙霞 任建科 

机构地区：[1]上海南方模式生物研究中心,上海201203

出　　处：《中国细胞生物学学报》2013年第3期316-321,共6页Chinese Journal of Cell Biology

基　　金：上海市自然科学基金(批准号:09ZR1422600)资助的课题~~

摘　　要：建立几丁质酶结构域内含蛋白1(chitinase domain containing 1,Chid1)基因剔除小鼠,观察小鼠表型和发育差异。设计了合适的基因剔除策略,成功构建了基因剔除打靶载体。以电穿孔方法将打靶载体导入ES细胞(embryonic stem cell),用G418和Ganciclovoir进行正负筛选,挑选抗药的阳性克隆,提取ES细胞基因组DNA,用长臂PCR鉴定出阳性ES细胞。将阳性ES细胞复苏培养后注入小鼠囊胚,获得嵌合体小鼠。嵌合体小鼠与C57BL/6J小鼠交配后获得Aguoti毛色的杂合子小鼠。在雌雄杂合子交配的后代中获得纯合子小鼠。从脑、脾脏、肝、肺的RNA水平鉴定来看,基因剔除小鼠的Chid1基因未表达,而杂合子、野生型小鼠有明显的该基因条带。经过初步的表型观察发现,Chid1基因剔除小鼠发育正常,未出现胚胎致死,交配繁殖能力无异常。几丁质酶结构域内含蛋白1(Chid1)基因剔除小鼠模型建立成功。Child1基因对于小鼠发育、生殖方面无明显作用。To generate chitinase domain containing 1 （Chidl） knockout mice, mice were observed in phe- notype and development. We designed an appropriate strategy and constructed the targeting vector. After electropo- ration of targeting vector into ES cells, positive clones were grown and selected using G418 and Ganciclovir, then injected into mouse blastulas to obtain chimeric mice. By mating with C57BL/6J mice, we got heterozygous mice with Aguoti coat. Then we got homozygous KO mice from heterozygousmice mating. RT-PCR results showed that Chidl expressed in brain, spleen, liver and lung from heterozygous and wild-type mice, but not in homozygous mice. In general, Chidl knockout mice develope normally and all mice are alive after birth, there is no defect in fertility and development, Chidl knockout mice are successfully generated and can be a good animal model for in- vestigate its biological function in vivo.

关 键 词：几丁质酶结构域内含蛋白1 基因打靶 基因剔除 

分 类 号：Q95-331[生物学—动物学]