中文摘要：

目的克隆Klf4基因并对其重组蛋白进行原核表达及分析。方法提取胎鼠皮肤mRNA后反转录为cDNA序列,用一对两端引入特定酶切位点引物,从该cDNA中扩增出Klf4基因编码区序列,将其克隆到pEasy-T3载体上。对质粒双酶切并回收其中Klf4基因片段后,克隆入pET-52b（＋）载体后转化Origmai B（DE3）型大肠杆菌,用IPTG诱导表达,最后采用SDS-PAGE对重组蛋白进行鉴定及分析。结果对所克隆的Klf4mRNA蛋白编码区的DNA序列分析表明,klf4CDS区包括终止密码子在内为1452 bp,与参照序列对比仅有四处存在差异,不仅其同源性达到99.72%,且其氨基酸序列同源性为100%;在IPTG诱导下pET-52b（＋）-Klf4重组质粒可表达与预期相符的约为57×103的蛋白质;经IPTG刺激后重组蛋白表达明显上调,其中IPTG为0.4 mol/L时效果最佳。结论从胎鼠皮肤中克隆的Klf4基因可在原核中表达。

英文摘要：

Objective As a member of Krüppel-like family of transcript factors,Klf4（Krüppel-like factor 4） plays an important and conditional role in cell proliferation,differentiation,apoptosis,embryo development and tumorigenesis in a cell content-dependent mode.The aim of this study was to clone Klf4 gene and analyzed its expression in E.coli,and to make an effort on the further study of its function.Methods Klf4 gene was cloned from the cDNA amplified from fetal mouse skin mRNA,and ligated with linear vector pEasy-T3.The recombinant plasmid was digested and segment with Klf4 gene was purified,then subcloned into prokaryotic expressing vector PET-52b（＋）.Results The recombinant plasmid was cloned into PET-52b（＋） vector,and transformed the E.coli strain Origmai B（DE3）.Its induced expression by IPTG was identified and analyzed by SDS-PAGE.Conclusion Klf4 gene cloned from the fetal mouse skin can be expressed in procaryotic organisms.The coding region of Klf4 in a length of 1452 bp and recombinant protein with 484 amino acids were successfully obtained just as anticipated.