小鼠LIF编码基因的克隆与真核表达载体pcDNA3.1-LIF的构建

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612 bp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LIF cDNA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。

Cloning of mouse full-lenth LIF gene and construction of eukaryotic expression vector 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 pcDNA3.1-LIF with neomycin resistant was constructed by homologous recombination.Results The amplfied fragments was LIF cDNA about 612 bp,it had 100% homogeneity with mouse LIF cDNA se...