摘要
本文应用差文库（ｓｕｂｔｒａｃｔｉｖｅｌｉｂｒａｒｙ）富集酒精性肝硬化的高表达基因。从酒精性肝硬化大鼠建立单方向ｃＤＮＡ基因文库，并提取单键ｃＤＮＡ。以４０倍的正常鼠肝ｍＲＮＡ标记生物素后，对肝硬化鼠肝的单链ｃＤＮＡ进行差杂交，滤掉表达相似序列。共得约９０个克隆，其中１０个克隆分析了约５００个碱基对。一个克隆的ＤＮＡ序列分析了８００个碱基对。查Ｇｅｎｅｂａｎｋ，证实为一新的基因。Ｎｏｒｔｈｅｒｎ印迹分析显示肝硬化时，其ｍＲＮＡ的表达增加数倍。
The effect of ethanol on hepatic gene expression has not been thoroughly investigated. To better understand the altered hepatic gene expression in alcoholic liver disease, we constructed a cDNA library from the liver of a rat which had been fed ethanol for 16 weeks using the intragastric ethanol feeding model (Tsukamoto-French Model). Liver histology demonstrated focal necrosis with periportal and pervenular fibrosis. A subtraction library was obtained by subtracting excess biotinylated normal rat liver mRNA from the alcoholic rat liver cDNA library. Approximately 90 colonies were obtained. Several colonies were selected for further study. One clone was sequenced in both directions using the dideoxynucleotide method. Comparison of this 800 base sequence with sequences in gene bank did not reveal a significent match. On northern analysis, this cDNA hybridized with a 1. 8 kd mRNA which was expressed at several fold higher levels in alcoholic rat liver compared with normal and with control-fed rat liver. In conclusion, we have isolated a cDNA clone which codes for a differentially expressed mRNA in alcoholic rat liver. This sequence has not been previously reported to gene bank.
出处
《胃肠病学和肝病学杂志》
CAS
1995年第1期17-19,共3页
Chinese Journal of Gastroenterology and Hepatology
关键词
基因文库
差文库
酒精性肝硬化
gene library subtractive library alcoholic cirrhosis.
