title: [Applying suppression subtractive hybridization technique to investigate gene differential expression profiles in rats liver during the course of heat stress].
abstract: OBJECTIVE
  To 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.
  
  
  METHODS
  There 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.
  
  
  RESULTS
  mRNAs 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.
  
  
  CONCLUSIONS
  Construction 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.
authors.affiliation: Department of Tropical Medicine, First Military Medical University, Guangzhou 510515, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Xiao
authors.forename: Fei
authors.initials: F
authors.lastname: Zou
authors.forename: Shao-Xi
authors.initials: SX
authors.lastname: Cai
mesh_terms: D000818:Animals; D018076:DNA, Complementary; D015870:Gene Expression; D018882:Heat Stress Disorders / Q000235:genetics*; D008099:Liver / Q000378:metabolism; D008297:Male; D009693:Nucleic Acid Hybridization / Q000379:methods*; D051381:Rats; D017207:Rats, Sprague-Dawley; D059467:Transcriptome
chemical_list: D018076:DNA, Complementary