【摘要】 目的克隆大鼠肝再生增强因子基因编码序列 ,并在原核细胞表达。方法按文献报道大鼠肝再生增强因子核苷酸序列合成引物 ,从 2周龄大鼠肝组织提取RNA ,利用RT PCR扩增大鼠肝再生增强因子基因编码区 ;将PCR扩增片段亚克隆到pBV2 2 1质粒 ,构建原核表达重组载体 ,导入到大肠杆菌后热诱导表达目的蛋白。结果从大鼠肝脏的RNA中扩增到长约 4 0 0bp的肝再生增强因子cDNA编码区基因 ,序列分析表明与文献报道一致 ;重组克隆经热诱导表达出分子量约 15kD大小的蛋白质 ,与预期值一致。结论从 2周龄大鼠肝组织中成功克隆肝再生增强因子基因 ,并获得了原核细胞高效表达。更多还原

【Abstract】 ObjectiveTo clone the gene of rat augmenter of liver regeneration (ALR),and express the protein encoded by ALR in the prokaryotic cells.MethodsThe coding region of ALR gene was amplified by a coupled one step reverse transcription PCR(RT PCR).The PCR fragment of ALR gene was subcloned to pBV221 plasmid,and constructed a prokaryotic expression vector,and then the recombinant plasmid was heat induced to express protein after transducted to E.coli.ResultsALR cDNA of 400bp was amplified successfully,which was consistent with the gene reported in literature after nucleotid sequence was detected.The specific expression band was observed in SDS PAGE analysis,and its moleculor weight was about 15 kD which was in accordance with the expecting value.ConclusionThe augementer of liver regeneration gene was cloned from the liver tissue of 2 week old SD rat and the protein encoded by the gene was expressed in the prokaryotic cells.更多还原