成骨细胞特异性表达Cre重组酶转基因小鼠建立.pdfVIP

HEREDITAS (Beijing) 2007 10 , 29(10): 1237―1242 ISSN 0253-9772 研究报告 DOI: 10.1360/yc-007-1237 Cre 程萱, 翁土军, 谭晓红, 侯宁, 王健, 林福玉, 黄培堂, 杨晓 , 100071 : 构建了含有骨钙素基因启动子、Cre 重组酶基因和人生长激素基因 polyA 的转基因载体pOC-Cre, 以显 微注射的方法将4.6 kb 的转基因片段OC-Cre 导入小鼠受精卵。16 只子代小鼠中经PCR 和Southern 杂交鉴定, 有 2 只小鼠携带外源基因, 整合率为 12.5%。为了检测OC-Cre 在转基因小鼠中表达的组织特异性, 将转基因首建 者小鼠与基因组上携带有LoxP 位点的条件性Smad4 基因敲除小鼠交配, PCR 结果显示, 仅在子代纯合型小鼠 骨组织基因组中扩增出了Cre 介导重组后的片段。将OC-Cre 转基因小鼠与ROSA26 报告小鼠交配, 利用LacZ 染色对双转基因阳性子代小鼠进行检测, 结果显示 Cre 重组酶在成骨细胞中特异性表达并介导 ROSA 基因座 LoxP 位点间的重组。所有这些结果说明：所建立的OC-Cre 转基因小鼠在成骨细胞中特异性表达Cre 重组酶, 并 能在体内介导成骨细胞基因组上LoxP 位点间的重组, 是一种理想的研制成骨细胞特异性基因敲除小鼠的工具 小鼠。 : Cre; 转基因小鼠; 成骨细胞; 显微注射 Establishment of osteoblast-specific Cre transgenic mice CHENG Xuan, WENG Tu-Jun, TAN Xiao-Hong, HOU Ning, WANG Jian, LIN Fu-Yu, HUANG Pei-Tang, YANG Xiao Genetic Laboratory of Development and Diseases, Beijing Institute of Biotechnology, Beijing 100071, China Abstract: An osteoblast-specific Cre transgenic construct (pOC-Cre) containing the osteocalcin promoter, Cre recombi- nase gene and polyA of human growth hormone gene was generated. The 4.6 kb DNA fragment of OC-Cre was introduced into fertilized zygotes by microinjection. 2 out of 16 offspring were identified as founders carrying the transgene by PCR and Southern blot analysis, and the integration efficiency is 12.5 %. To check the tissue distribution of the OC-Cre, the OC-Cre transgenic founder mice were bred with the mice carrying Smad4 conditional alleles. PCR results showed that the genomic DNA fragments after Cre m