逆转录病毒转导的重组酶Cre在体外对LoxP标记的小鼠骨髓细胞DNA重组  

Recombination of LoxP-floxed gene by Cre recombinase expressed by retrovirus system in vitro

作　　者：陆群[1] 吴补领[2] 周学东[1] 王冀姝[2] 韩骅[2] 

机构地区：[1]四川大学华西口腔医学院,四川成都610041 [2]第四军医大学基础部医学遗传学与发育生物学教研室,陕西西安710033

出　　处：《第四军医大学学报》2003年第9期785-787,共3页Journal of the Fourth Military Medical University

基　　金：全国高等学校骨干教师资助计划

摘　　要：目的 :研究逆转录病毒体系转导的重组酶Cre在体外对LoxP标记的基因组进行重组的能力 .方法 :用逆转录病毒表达Cre,体外感染条件性基因剔除 (即LoxP标记的基因 )小鼠来源的原代培养的骨髓细胞 .从被感染的骨髓细胞中提取基因组DNA ,利用PCR的方法显示LoxP标记的基因组长度的剪切前后改变 ,从而反映Cre在体外对LoxP标记的基因组的重组力 .结果 :被表达有Cre的逆转录病毒感染后的骨髓细胞 ,其特定的基因组长度发生了预期的变化 ,被标记的片段部分发生缺失 .结论 :在体外 ,用逆转录病毒体系转导重组酶Cre能够有效地对LoxP标记的基因组进行重组 .AIM: To study the recombination LoxP floxed gene by Cre recombinase expressed by retrovirus system in vitro . METHODS: Cre recombinase was expressed by retrovirus system, the bone marrow cells which containing LoxP floxed gene from conditional gene knock out mice were infected, genome DNA from the bone marrow cells infected by retrovirus was extracted and the changes of the LoxP floxed gene length were observed by PCR. RESULTS: LoxP floxed gene of the cells infected by retrovirus was deleted. CONCLUSION: Cre recombinase expressed by retrovirus system has high recombinase activity in vitro . This system is helpful in further studying the functions of specific gene.

关 键 词：CRE重组酶 条件性基因剔除 逆转录病毒 

分 类 号：R595.1[医药卫生—内科学]