摘要
建立 FBXL15 基因条件型敲除小鼠模型,为研究该基因所发挥的重要生理功能提供材料和思路。采用KO first策略,构建打靶载体,将1stloxP插入1~2号内含子,在3~4号内含子之间插入FRT-SA-IRES-lacZ-loxP-neo-loxP元件,通过Long Range PCR及Southern blot筛选出中靶克隆,随后进行囊胚注射,并将发生同源重组的ES细胞注射进C57BL/6J小鼠囊胚,移入受体小鼠子宫,最后将得到的嵌合体雄鼠与C57BL/6J雌鼠交配获得FBXL15-LoxP小鼠。PCR结果显示FBXL15的Loxp小鼠模型构建成功,该模型可为进一步研究FBXL15在胚胎发育和骨代谢中的调控作用提供工具。
To generate F-box and Leucine-rich repeat protein 15 conditional knockout mice model,and confirm the genotype for further functional research. Using KO first strategy,to construct the conditional knockout vector targeting exon2 and exon3.In additional, FRT-SA-IRES-lacZ-loxp-neo-loxp element between intron 3 and intron 4 was also inserted.By Long Range PCR and Southern blot methods the positive clones were selected. Therefore the dual selected ES cells were microinjected into blastula of C57BL/6J mice, then blastula transplantations into the host mice,Chimerical mice were generated and the FBXL15-Lox Ptargeted mice were identified by PCR method. This animal model established the foundation for further study of FBXL15＇s pivotal roles in regulation upon embryos development and bone metabolism
出处
《中国生物工程杂志》
CAS
CSCD
北大核心
2015年第4期74-79,共6页
China Biotechnology
基金
国家自然科学基金资助项目(2011CB910602
2012CB910702)
