摘要
白蛋白mRNA已被证实是反映白蛋白合成变化的重要指标。实验探讨大鼠白蛋白基因片段的克隆及白蛋白mRNA半定量检测方法的建立。使用一步法逆转录聚合酶链反应扩增的白蛋白cDNA片断 ,连接入pGEM T载体中 ,经ABIPRISM 3 77型全自动核苷酸分析仪检测。同时用不同浓度的内毒素处理肝细胞后检测其白蛋白mRNA的表达。扩增的基因片段经DNA序列分析 ,证实其由40 1个碱基组成。同时发现 ,内毒素可以抑制白蛋白mRNA的表达 ,且抑制作用与内毒素的剂量相关。提示 。
Objective To clone and analyze rat albumin gene. Methods Total RNA was prepared from rat hepatocytes. cDNA fragment encoding rat albumin was amplified by RT-PCR, and then was cloned into pGEM-T vector. Inserted rat gene was sequenced by ABI PRISM 377DNA Sequencer. Meanwhile, the albumin mRNA expression was observed in hepatocytes treated with 1ng/ml and 1g/ml LPS. Results Rat albumin gene was successfully cloned. The expression of albumin mRNA was inhibited by LPS, and the inhibition was dose dependent. Conclusion The cloned rat albumin gene was composed of 401 base pairs and had 100% nucleotide homology with that reported in Genbank. The albumin expression was decreased in hepatocytes treated with LPS. A stable method of semi-quantitative analysis of rat albumin mRNA was established.
出处
《解放军医学杂志》
CAS
CSCD
北大核心
2003年第7期611-612,共2页
Medical Journal of Chinese People's Liberation Army
基金
全军医学科研"十五"计划重点项目 (编号 0 1Z0 1 1 )
江苏省青年科技基金 (编号BQ2 0 0 0 0 1 4 )资助课题
关键词
白蛋白
基因表达
克隆
逆转录聚合酶链反应
albumin
gene expression, clone
reverse transcriptase polymerase chain reaction
