MOLECULAR CLONING AND SEMI-QUANTITATIVE ANALYSIS OF RAT ALBUMIN GENE
- VernacularTitle:大鼠白蛋白基因片段的克隆及其mRNA半定量检测方法的建立
- Author:
Xinying WANG
;
Weiqin LI
;
Ning LI
- Publication Type:Journal Article
- Keywords:
albumin;
gene expression, clone;
reverse transcriptase polymerase chain reaction
- From:
Medical Journal of Chinese People's Liberation Army
2001;0(07):-
- CountryChina
- Language:Chinese
-
Abstract:
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.
