人肝再生增强因子的克隆及表达

Cloning and Expression of Human Augmenter of Liver Regeneration

根据同源克隆的原则，依照大鼠ALRcDNA编码区两端核苷酸序列合成一对引物，以人胎肝细胞mRNA的RT产物为模板扩增出一条380bpcDNA片段，测序结果表明与大鼠ALR有85％同源性，将人ALRcDNA插入PBV220质粒转化E．coliJM109感受态细胞，筛选到的阳性克隆经温度诱导后，SDS-PAGE结果表明目的蛋白含量占菌体总蛋白的25％左右。

A 38ohp cDNA fragment was amplified using the reverse transcriptionproduct of human fetal liver cell mRNA as template. The primers were designed according to thenucleotid sequence of 3' and 5' ends of coding region of rat augmenter of liver regeneration(ALR) cDNA. Sequencing proved that 85% of the sequences of the fragments werehomologous to those of rat ALR. The huamn ALR cDNA was inserted into plashhed PBV220 andtransformed to permissive E. colt JM109 cell, then the positive clones were screened out andinduced by temperature. SDS-PAGE proved that about 25% of total somatic protein wereexpressed.