基于pMTR1质粒的转基因小鼠突变研究模型的建立  

Development of pMTR1 plasmid-based transgenic mice model for studying gene mutation in vivo

作　　者：卢洋[1] 黎怀星[1] 李建秀[2] 傅继梁[1] 

机构地区：[1]第二军医大学基础医学部医学遗传学教研室,上海200433 [2]第二军医大学基础医学部细胞生物学教研室,上海200433

出　　处：《第二军医大学学报》2000年第6期537-540,共4页Academic Journal of Second Military Medical University

基　　金：国家自然科学基金!资助项目 (395 70 40 0 );军队"九五"重点攻关课题!基金资助项目 (96 Z0 33)

摘　　要：目的 :建立在基因组中整合有 p MTR1质粒的 C5 7BL / 6转基因小鼠家系 ,为体内基因突变研究提供有效的动物模型。方法 :利用分子克隆技术构建 p MTR1质粒。将其线性化后 ,用显微注射法注射入 5 72只 C5 7BL / 6小鼠受精卵 ,再将它们分别移入 45只受体母鼠的输卵管中 ,共产仔 44只 ,存活 35只 ,采用 PCR,PCR- Southern和基因组 Southern杂交三级筛选法鉴定子代小鼠。以 4只经基因组 Southern杂交证实在基因组 DNA中整合有多拷贝结构完整的 p MTR1质粒的小鼠作Founder小鼠 ,进行转基因小鼠的建系工作。对每个 Founder小鼠已有的 F1代及 F2代转基因小鼠的基因组 DNA进行p MTR1质粒回收实验。结果 :从转基因小鼠基因组中可以回收出结构、功能完整的 p MTR1质粒。结论 :(1)建立了在基因组 DNA中整合有 p MTR1质粒的 C5 7BL / 6转基因小鼠家系 ;(2 )Objective: To establish C57BL/6 transgenic mice lineages containing pMTR1 plasmids as an efficient animal model for studying gene mutation in vivo . Methods: The linearized pMTR1 DNAs, constructed by molecular cloning, were injected into 572 fertilized eggs of C57BL/6 mice by microinjection. The manipulated embryos were transferred into the oviducts of 45 pseudopregnant mice respectively, from which 35 offsprings were obtained. The genomic DNAs of these offsprings were analyzed with PCR and genomic Southern blotting. Four mice whose genomes integrated with copies of intact pMTR1 vectors were chosen as founders to establish transgenic mice lineages. The recovery of pMTR1 plasmid were conducted on the genomic DNAs of F1 or F2 transgenic offsprings. Results: It was showed that intact and functional pMTR1 plasmid could be efficiently recovered from the genomic DNAs of these mice. Conclusion: (1) The transgenic mouse lineages containing copies of a stably integrated pMTR1 plasmid is established. (2) The transgenic mice are suitable models for studying gene mutation in vivo .

关 键 词：转基因小鼠 基因突变 PMTR1质粒 

分 类 号：R392.2[医药卫生—免疫学]