【摘要】 对利用基因诱捕技术进行小鼠基因剔除做了初步的探索 ,为进一步应用该技术进行小鼠基因功能研究奠定了基础 .利用基因诱捕载体转染小鼠ES细胞 ,获得了 36株neo基因单拷贝整合的诱捕ES细胞 ,其中 14株细胞表达有活性的 β半乳糖苷酶 .将 3株诱捕ES细胞分别经显微注射引入到受体囊胚中 ,再植入假孕母鼠的子宫中使其发育成小鼠 .两株细胞得到了程度不同的嵌合体小鼠 ,其中一株诱捕ES细胞整合至生殖系 .利用质粒拯救实验获得了诱捕载体整合位点附近的基因组序列 ,通过序列比对发现被诱捕的基因可能是一个新基因 .X gal染色结果显示 ,该基因的表达局限于小鼠腹部及肢芽的部位 .更多还原

【Abstract】 A gene trap construct was used to transfect mouse ES cells. 36 ES colonies trapped by one copy of neo gene were obtained. The β-galactosidase activity was detectable in 14 ES colonies. ES cells from 3 trapped lines were introduced into blastocysts by microinjection. Two chimeric mice lines were generated. One trapped mutation went through the germline. Genomic sequences adjacent to integration sites of the constructs were isolated by plasmid rescue. The results of sequencing suggested that the trapped gene is possibly a novel gene. The expression pattern of this gene shown by β-galactosidase expression was restricted in abdomen and lib bud of E10 5 mouse embryo.更多还原