CLONING AND EXPRESSION ANALYSIS OF SEVERAL Beauveria bassiana GENES INVOLVED IN ALKANE degradation.

NICOLÁS PEDRINI AND M. PATRICIA JUAREZ

Córdoba

Congreso; 44 Reunión Anual- Soc. Argentina de Investigac Bioquim. y Biol. Mol.; 2008

Soc. Arg. Investigac Bioquim.

BIOCELL 32 (Suppl.), pp. 32, 2008 CLONING AND EXPRESSION ANALYSIS OF SEVERAL Beauveria bassiana GENES INVOLVED IN ALKANE DEGRADATIONGENES INVOLVED IN ALKANE DEGRADATION 2 2 1 Pedrini NL , Keyhani NO , Juarez MP .Pedrini NL , Keyhani NO , Juarez MP . 1 2 INIBIOLP (CONICET-UNLP), Argentina. University of Florida, USA. E-mail: nicopedrini@yahoo.comINIBIOLP (CONICET-UNLP), Argentina. University of Florida, USA. E-mail: nicopedrini@yahoo.com Entomopathogenic fungi have the ability to degrade insect cuticular hydrocarbons and to utilize them for energy production and incorporation into cellular components. The first oxidation round is carried out by a cytochrome P450 (P450alk), after successive steps the corresponding metabolites will eventually provide the appropriate fatty acyl-CoA for complete degradation in the peroxisomes, the site of b-oxidation in fungi. Eight gene fragments displaying high homology to cytochrome P450alk as well as the peroxisomal enzymes catalase and acyl CoA oxidase were identified in a B. bassiana expressed sequence tagged (EST) collection. Full-length sequences for each gene were isolated by 5’ and 3’-rapid amplification of cDNA ends (RACE). Expression analysis of the genes by quantitative real-time RT PCR using fungal cells grown on n-hexadecane (C16), n-eicosane (C20), ntetracosane (C24) or n-octacosane (C28) revealed overlapping but differential expression of the isolated genes. These data indicate thatb-oxidation in fungi. Eight gene fragments displaying high homology to cytochrome P450alk as well as the peroxisomal enzymes catalase and acyl CoA oxidase were identified in a B. bassiana expressed sequence tagged (EST) collection. Full-length sequences for each gene were isolated by 5’ and 3’-rapid amplification of cDNA ends (RACE). Expression analysis of the genes by quantitative real-time RT PCR using fungal cells grown on n-hexadecane (C16), n-eicosane (C20), ntetracosane (C24) or n-octacosane (C28) revealed overlapping but differential expression of the isolated genes. These data indicate thatB. bassiana expressed sequence tagged (EST) collection. Full-length sequences for each gene were isolated by 5’ and 3’-rapid amplification of cDNA ends (RACE). Expression analysis of the genes by quantitative real-time RT PCR using fungal cells grown on n-hexadecane (C16), n-eicosane (C20), ntetracosane (C24) or n-octacosane (C28) revealed overlapping but differential expression of the isolated genes. These data indicate that B. bassiana is likely to contain multiple hydrocarbon degradative pathways with overlapping substrate specificities.is likely to contain multiple hydrocarbon degradative pathways with overlapping substrate specificities.