Cloning a novel catalase gene of Sporothrix schenckii with degenerate PCR and RACE

Abstract

Abstract: Objective To isolate a novel catalase homologous gene from yeast-form Sporothrix schenckii and to make a designation.Methods Oligonucleotide primers were designed according to the conserved areas of the other 7 fungal catalase genes.Partial Sscat cDNA was amplified by PCR,and special primers were designed by RACE method to amplify the 3′cDNA and 5′cDNA.Results The full-length Sscat cDNA sequence was 1746 bp with an open reading frame of 1500 bp encoding 499 amino acids.The predicted molecular mass of Sscat was 56.07 kDa.The deduced amino acid sequence of Sscat showed 66.3% and 56.6% identity with those of Aspergillus oryzae and A.clavatus.An intron was identified within the 933-1063 bp Sscat genomic DNA sequence of S.schenckii.Conclusions Degenerate PCR combined with RACE is effective in searching and isolating novel genes of S.schenckii.

Key words: Sporothrix schenckii, catalase, degenerate PCR, RACE

CLC Number:

R379.9

WANG Xiao-hui, LIU Wei, LI Ruo-yu. Cloning a novel catalase gene of Sporothrix schenckii with degenerate PCR and RACE[J]. Chinese Journal of Mycology, 2011, 6(5): 271-275.

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