AIMTo explore the molecular mechanism of heat stress by isolating differentially expressed genes occurred during the course of heat stress and filtrating functional gene segments concerned with heat stress with Suppression Subtractive Hybridization technique.

METHODSThere were 2 groups in the experiment: heat stress group and normal temperature control group. mRNAs were isolated from the liver tissue individually and then transcripted into cDNAs. After cut with endonuclease and ligation with adaptors, SSH was carried out with cDNAs of heat stress group as tester while cDNAs of control group as driver. Products were cloned into T/A vector to form recombined plasmids to transfect competence cells for filtration and appraisal.

RESULTSmRNAs were abundant and of high quality, cDNAs were reverse-transcripted and then cut into segments about 500 bp successfully. Ligation efficiency was satisfied. The subtractive hybridization library was constructed successfully after effective hybridization and 180 segments were primarily isolated from it.

CONCLUSIONConstruction of heat stress differentially expressed genes subtractive library has established the foundation of filtrating heat stress concerned genes, it is of active significance for exploring functional genes that controlled heat stress.