学术
小鼠LRP16基因同源重组ES细胞克隆的鉴定
韩为东[1] 伍志强[1] 赵亚力[1] 司艺玲[1] 母义明[2] 孟元光[3] Masatoshi Nomura[4]
[1]解放军总医院分子生物室,中国北京100853 [2]解放军总医院内分泌科,中国北京100853 [3]解放军总医院妇产科,中国北京100853 [4]九州大学医学院第三内科,日本福岗812-8582
摘 要:
目的：为深入研究LRP16基因的生理功能，本研究将小鼠LRP16基因特异打靶载体pBA16转染小鼠胚胎干细胞（embryonic stem cell），并筛选同源重组克隆。方法：通过电转染方法将LRP16插入失活型打靶载体导入ES细胞，经G418筛选，挑取抗性克隆，Southern blot方法鉴定同源重组ES细胞克隆。结果：打靶载体成功转染ES细胞，Southern blot筛选100个抗性克隆，结果显示有一个打靶序列同源重组型插入的Es细胞克隆。结论：筛选到同源重组型ES细胞，为下一步建立LRP16缺失型小鼠并为从整体水平研究LRP16基因的生理功能奠定基础。[著者文摘]
文章出处:
《军医进修学院学报》-2006年27卷6期 -406-408页
分 类 号:
文献标识码:
A
文章编号:
1005-1139（2006）06-0406-03
Identification of homologously recombinant clone of ES cells for mouse LRP16 gene
HAN Wei-Dong, WU Zhi-Qiang , ZHAO Ya-Li, SI Yi-Ling, MU Yi-Ming, MENG Yuan-Guan, Masatoshi Nomura （ 1. Research Laboratory of Molecular Biology, PLA General Hospital, Beijing 100853, China; 2. Endocrinology, PLA General Hospital, Beijing 100853, China; 3. Gynaecology and Obstetrics, PLA General Hospital, Beijing 100853, China; 4. Kyushu University, Japan, 812-8582）
Abstract:
Objective：Previous studies have demonstrated that LRP16 is an estrogen-responsive gene, the expression level of which is tightly linked with proliferation and invasive growth of human breast cancer ceils. To further explore the physiological function of LRP16, here, we transfected LRP16 targeting vector pBA16 into ES ceils and screened the ES clone with inactive LRP16 gene. Methods： ES ceils were screened with G418 and the homologously recombinant clone was identified from 100 clones by southern blot analysis. Results：There was an ES clone with targeting sequence homolougously inserted. Conclusion： A homologous recombinant was obtained. The study lays the foundations of preparing mouse models of LRP16 knockout.[著者文摘]
Key words:
LRP16 ; gene targeting; embryonic stem cells
基金资助:
国家自然科学基金资助项目（30471813）,北京市自然科学基金资助项目（7052061,5052024）
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