Sumf2条件性基因剔除小鼠模型的建立及初步表型分析  

Generation of Sum f2 Conditional Gene Knockout Mouse Model and Its Preliminarily Phenotypic Analysis

作　　者：匡昉哲[1] 庄华 周涛[1] 匡颖 王铸钢[1,2] 

机构地区：[1]上海交通大学医学院,上海200025 [2]上海南方模式生物研究中心,上海201203

出　　处：《实验动物与比较医学》2013年第6期425-432,共8页Laboratory Animal and Comparative Medicine

基　　金：上海市科委项目资助（11DZ2292400）,国家“863”计划重大项目（2007AA022185）

摘　　要：目的建立Sumf2条件性基因剔除小鼠模型，为整体水平研究Sumf2基因的生物学功能创造条件。方法运用生物信息学，采用ET克隆、同源重组、显微注射等技术成功建立Sumf2条件性基因剔除小鼠模型，并通过该模型与Ella-cre转基因小鼠交配，获得Sumf2基因剔除小鼠模型，通过PCR、RT-PCR、WesternBlot等方法在不同水平验证Sumf2基因的表达；对该小鼠模型进行初步的表型分析。结果经胚胎干细胞打靶，获得19个正确同源重组的克隆，重组效率为20％：阳性克隆经囊胚注射，获得2只嵌合体雄鼠；嵌合雄鼠与C57BL／6J小鼠交配，获得28只灰鼠，经PCR鉴定16只为杂合子小鼠，阳性率为57％；与Ella—ere转基因小鼠交配，获得了Sumf2基因剔除小鼠模型，DNA、RNA及蛋白水平证明该基因已成功剔除；初步的表型观察未发现Sumf2基因剔除小鼠生长发育、血常规及血生化指标等出现异常改变。结论成功建立Sumf2条件性基因剔除小鼠模型，该基因纯合缺失未发现明显的生长发育异常，为进一步的基因功能研究奠定了基础。Objective To establish the sulfatase modifying factor 2 （Sumf2） gene conditional knockout （CKO） mouse model and to analyze the biological functions for further in vivo study. Methods Use the technology of bioinformatics, ET cloning, homologous recombination, and microinjection, etc to establish the CKO mouse model. Mating transgenic （TG） homozygous EIIa-cre males with CKO heterozygous females, Sumf2 gene knock out （KO） mouse model was established. The deletion of Sumf2 gene in the KO mouse was confirmed by PCR, RT-PCR and western-blot analysis. Results There are about 20% - 19 ES cells positive clones, 2 chimeras males and about 57% of 28 Aguoti mice-16 heterozy- gous obtained. Based on this result, mating TG homozygous EIIa-cre males and CKO heterozygous females, Sumf2 gene KO mouse model has been established. Analyzed by PCR, RT-PCR and Western Blot, it is verified for the deletion of Sumf2 gene at different levels. Preliminarily phenotypic observations show that the growth and development of Sumf2 KO mice were generally normal, and abnormalities were not found from the data of routine blood tests and some of serum biochemical tests in comparison with those of wild type mice. Conclusions The establishment of Sumf2 CKO mouse model and Sumf2 KO mouse model has paved the way for further study of the functions of this gene.

关 键 词：Sumf2基因 条件性基因剔除小鼠 表型分析 

分 类 号：Q95-33[生物学—动物学]