小鼠LIF编码基因的克隆与真核表达载体pcDNA3．1-LIF的构建  

Cloning of mouse full-lenth LIF gene and construction of eukaryotic expression vector pcDNA3.1-LIF

作　　者：姜秋[1] 艾永华[1] 聂代邦[2] 孙宏晨[1] 欧阳红生[2] 

机构地区：[1]吉林大学口腔医院儿童牙病科,吉林长春130041 [2]吉林大学畜牧兽医学院生物技术系,吉林长春130062

出　　处：《吉林大学学报（医学版）》2008年第5期782-785,F0002,共5页Journal of Jilin University：Medicine Edition

基　　金：吉林省科技厅科技发展计划项目资助课题(200705347)

摘　　要：目的:通过克隆小鼠LIF编码基因,构建pcDNA3.1-LIF真核表达载体,为下一步建立转基因动物模型奠定基础。方法:采用ICR雌性小鼠子宫组织,提取总RNA,运用RT-PCR技术,扩增出小鼠LIF基因全长编码序列,采用同源重组方法构建具有新霉素抗性的小鼠LIF真核表达载体pcDNA3.1-LIF。结果:通过PCR获得了约612bp大小的特异性cDNA片段。经核苷酸序列分析,扩增得到的片段与小鼠LIFcDNA序列同源性为100%。经PCR及酶切鉴定筛选阳性克隆菌,表明LIF编码序列正确连入pcDNA3.1载体中。结论:成功克隆了小鼠LIF编码基因,并构建了真核表达载体pcDNA3.1-LIF。Objective To construct an eukaryotic expression vector pcDNA3.1-LIF by cloning mouse LIF genes and provide basis for establishment of transgenic animal models. Methods Total RNA was extracted from the tissue of the ICR mouse uterus . The full-lenth LIF gene of the mouse was amplified by RT-PCR and. an eukaryotic expression vector peDNA3.1-LIF with neomycin resistant was constructed by homologous recombination. Results The amplfied fragments was LIF eDNA about 612 bp, it had 100% homogeneity with mouse LIF cDNA sequence by nueleotide sequence analysis. An eukaryotie expression vector pcDNA3. 1-LIF with neomycin resistant of the mouse was successfully constructed by the identification of enzyme digestion. Conclusion The mouse LIF gene is successfully cloned and an eukaryotic expression vector pcDNA3. 1-LIF is successfully constructed.

关 键 词：白血病抑制因子 聚合酶链反应 基因克隆 真核表达载体 小鼠 近交ICR 

分 类 号：Q782[生物学—分子生物学]