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http://dx.doi.org/10.7852/ijie.2014.28.2.71

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

Liu, Ya-Qi (Department of Life Science & Environmental Biochemistry, College of Natural Resources and Life Science, Pusan National University)
Park, Nam Sook (Department of Life Science & Environmental Biochemistry, College of Natural Resources and Life Science, Pusan National University)
Kim, Yong Gyun (Department of Life Science & Environmental Biochemistry, College of Natural Resources and Life Science, Pusan National University)
Kim, Keun Ki (Department of Life Science & Environmental Biochemistry, College of Natural Resources and Life Science, Pusan National University)
Park, Hyun Chul (Department of Life Science & Environmental Biochemistry, College of Natural Resources and Life Science, Pusan National University)
Son, Hong Joo (Department of Life Science & Environmental Biochemistry, College of Natural Resources and Life Science, Pusan National University)
Hong, Chang Ho (Department of Life Science & Environmental Biochemistry, College of Natural Resources and Life Science, Pusan National University)
Lee, Sang Mong (Department of Life Science & Environmental Biochemistry, College of Natural Resources and Life Science, Pusan National University)
Publication Information
International Journal of Industrial Entomology and Biomaterials / v.28, no.2, 2014 , pp. 71-84 More about this Journal
Abstract
The full-length heat shock protein 88 (HSP88) complementary DNA (cDNA) of Paecilomyces tenuipes 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) of 2,139-basepair encoding 713 amino acid residues. The deduced amino acid sequence of the P. tenuipe s Jocheon-1 HSP88 cDNA showed 77% identity to Nectria haematococca HSP88 and 45-76% identity to other fungal homologous HSP88s. Phylogenetic analysis and BLAST program analysis confirmed that the deduced amino acid sequences of the P. tenuipes Jocheon-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-terminal region. 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 conditions for the growth of the entomopathogenic fungus, mRNA expression of P. tenuipes Jocheon-1 HSP88 was quantified by real time PCR (qPCR). The results showed that heat shock stress induced a higher level of mRNA expression compared to normal growth conditions.
Keywords
Paecilomyces tenuipes; heat shock protein 88; HSP88 cDNA; baculovirus-infected insect Sf9 cells;
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