Molecular Cloning of the cDNA of Heat Shock Protein 88 Gene from the Entomopathogenic Fungus, Paecilomyces tenuipes Jocheon-1

The full-length heat shock protein 88 (HSP88) complementary DNA (cDNA) of Paecilomycestenuipes Jocheon -1 was obtained by screening the Paecilomyces tenuipes (P. tenuipes) Jocheon-1 Uni-Zap cDNA library and performing 5' RACE polymerase chain reaction(PCR). The P. tenuipes Jocheon-1 HSP88 cDNA contained an open reading frame (ORF) of2,139-basepair encoding 713 amino acid residues. The deduced amino acid sequence of theP. tenuipe s Jocheon-1 HSP88 cDNA showed 77% identity to Nectria haematococca HSP88and 45-76% identity to other fungal homologous HSP88s. Phylogenetic analysis and BLASTprogram analysis confirmed that the deduced amino acid sequences of the P. tenuipesJocheon-1 HSP88 gene belonged to the ascomycetes group within the fungal clade. The P. tenuipes Jocheon-1 HSP88 also contained the conserved ATPase domain at the N-terminalregion. The cDNA encoding P. tenuipes Jocheon-1 HSP88 was expressed as an 88 kilodalton(kDa) polypeptide in baculovirus-infected insect Sf9 cells. Under higher temperature conditionsfor the growth of the entomopathogenic fungus, mRNA expression of P. tenuipes Jocheon-1HSP88 was quantified by real time PCR (qPCR). The results showed that heat shock stressinduced a higher level of mRNA expression compared to normal growth conditions. KCI Citation Count: 0