Construction and evaluation of subtractive libraries in association with heat adaptation differentially expressed genes in the rat liver
- VernacularTitle:大鼠肝细胞热适应差异表达基因消减文库的构建与鉴定
- Author:
Jun XIAO
;
Fei ZOU
;
Shaoxi CAI
;
Yue JIN
- Publication Type:Journal Article
- Keywords:
Heat adaptation;
Genes;
Suppression subtractive hybridization;
Rats
- From:
Chinese Journal of Pathophysiology
1989;0(06):-
- CountryChina
- Language:Chinese
-
Abstract:
AIM: To construct subtractive libraries in association with heat adaptation differential expressed genes. METHODS: The experiment was carried out with heat adapted rat model and normal temperature control. mRNA was extracted from liver tissue and reverse-transcripted into cDNA. After cut and ligation, suppression subtractive hybridization was executed with each group of cDNA as tester and the other one as driver to construct subtractive libraries. Specificity of the libraries was evaluated by approach of comparing G3PDH gene PCR products with template from the library and unsubtractive sample. Reliability of the libraries was evaluated by primarily isolation and screening. RESULTS: PCR results confirmed that G3PDH concentration was greatly reduced compared with control group, which suggested that specificity of the libraries was high. 300 target segments were isolated from the library, 27 of them were verified to be differential expressed genes, which suggested that the libraries were reliable and efficient. CONCLUSION: This study founded the basis of further investigation on heat adaptation mechanism by approach of constructing subtractive libraries in association with heat adaptation differential expressed genes.
