小鼠LRP16基因打靶载体的构建  

Construction of LRP16 gene targeting vector

作　　者：伍志强[1] 韩为东[1] 孟元光[2] 司艺玲[1] 赵亚力[1] 母义明[3] Masatoshi Nomura 

机构地区：[1]解放军总医院分子生物学室,北京100853 [2]解放军总医院妇产科,北京100853 [3]解放军总医院内分泌科,北京100853 [4]日本九州大学医学院医学与生物调控实验室

出　　处：《军事医学科学院院刊》2006年第4期315-318,共4页Bulletin of the Academy of Military Medical Sciences

基　　金：国家自然科学基金(30471813;30471990);北京市自然科学基金(7052061)资助项目

摘　　要：目的:为利用基因敲除技术进行LRP16基因生理功能的研究,本研究构建了针对小鼠LRP16基因的打靶载体。方法:通过PCR方法筛选129品系小鼠基因组文库中LRP16克隆,在第5外显子内插入SA-R IES-βgeo序列,构建插入失活型打靶载体。结果:将包含第5至11外显子的LRP16基因组片段亚克隆到pB luescript SKⅡ+载体,SA-IRES-βgeo序列正确插入第5外显子中。结论:成功构建了第5外显子插入失活型LRP16基因打靶载体,为下一步筛选同源重组ES细胞奠定基础。Objective： To construct a LRP16 gene targeting vector. Methods： The targeting sequence of LRP16 gene was obtained from 129 mouse genomic library by means of polymerase chain reaction screening. The SA-RIES-β geo fragment was designed and inserted into the LRP16 fifth exon to inactivate LRP16. Results： The LRP16 fragment,including exons 5 to 11 ,was then subcloned into the pBluescript SK Ⅱ^＋ vector. The restriction map demonstrated that the SA-IRES-β geo fragment was correctly inserted into the LRP16 fifth exon. Conclusion： A LRP16 gene targeting vector was successfully constructed, The study facilitates the screening of the homologous recombinant.

关 键 词：LRP16 基因打靶 载体 

分 类 号：Q78[生物学—分子生物学]