摘要
目的 采用RT -PCR一步法 ,克隆人肝再生增强因子 (hALR)编码序列。方法 按文献报道人肝再生增强因子核苷酸序列合成引物 ,提取人胎肝组织总RNA ,利用一步RT -PCR法扩增人肝再生增强因子编码区基因 ;将PCR扩增片断亚克隆到pBV2 2 1质粒 ,构建原核表达载体。结果 获得长约 40 0bp的hALPcDNA编码区基因 ,序列分析表明与文献报道一致。结论 通过RT -PCR一步法成功克隆人肝再生增强因子基因 。
Objective To clone the gene of human augmenter of liver regeneration(ALR) by using a coupled one-step reverse transcription PCR procedure.Methods Total RNA was extracted from the liver tissue of 4-month-ole fetus. The coding region of ALR cDNA was amplified by a coupled one-step reverse transcription PCR. According to the nuclear acid sequence of human ALR cDNA, three primers of P1, P2and P3 were designed to clone and identify the ALR gene. The PCR fragment was subcloned to pBV221 plasmid, and constructed a prokaryotic expression vector.Results ALR cDNA of 400bp was amplified successfully, which was consistent with the gene reported in literature after nucleotide sequence was detected.Conclusions The human augmenter of liver regeneration gene was cloned, which provided a sound basis for attaining the product of genetic engineering and studying many kinds of biological functions of human ALR.
