摘要
目的分离酒精性肝硬化病人的高表达基因 E251,并研究它的表达.方法与结果从我们以前在酒精性肝硬化大鼠中分离到的一个新基因 E251中,设计一对引物用逆转录 PCR 法从酒精性肝硬化病人的肝组织中分离到人肝 E251基因,DNA 序列对比证实与大鼠肝的 E251为同源基因,有大于95%的同源性.它的 mRNA 表达在酒精性肝硬化病人中比正常人高约2倍,说明在酒精刺激下,E251基因的表达增强.人肝 E251约0.95Kb 大小,含有 AATAAA的特别序列,并含有两个蛋白激酶 C 磷酸化激活位点.结论 1.存在与大鼠 E251同源的人类 E251基因;2.在酒精性肝硬化病人中,E251的表达增强.
Objective The aim of this study is to isolate a human homologue to E251(Rat),a previously unreported cDNA that we isolated from rat cDNA library by subtraction hybridization(Clinical Research 1994,42:241A).Methods and Results Two 20-mers approximately 500 base pairs on the rat E251 cDNA was synthesized.Reverse Transcription-Polymerase Chain Reaction was carried out using these paimers and total RNA from human alcoholic liver was as template.A single band of the appropriate size was obtained and inserted into PCR script(Stratagene). This was sequenced using the dideoxynucleotide method.The 500 nucleotide insert had approximately 95% homolgy with rat E251.The cDNA codes a histine-rich peptide as well as two consensus activation sites for protein kinasc C phosphorylation.Northern blot analysis of human liver revealed a single band of approximately 0.95 kb and alcoholic cirrhotic liver contained approcimately twice as much E251 mRNA as did normal human liver. Conclusion There was a human homologue gene increased expression in human alcoholic cirrhosis to rat E251.
出处
《中华肝脏病杂志》
CAS
CSCD
1997年第1期22-23,共2页
Chinese Journal of Hepatology
