Biotechnology1682-296x1682-2978Asian Network for Scientific Information10.3923/biotech.2009.194.203Trichoderma virens UKM1]]>OhS.S.L. BakarF.D.A. AdnanA.M. MahadiN.M. HassanO. MuradA.M.A. 2200982In this study, the isolation and characterization of Trichoderma virens glyceraldehyde-3-phosphate dehydrogenase gene (GPD1) and its promoter is described. A cDNA clone of a partial GPD1 had been identified from an ongoing work on T. virens Expressed Sequence Tag (EST) analysis. This led to the isolation of a 2.9 kb T. virens GPD1 that encompasses the 5-regulatory flanking region (1,364 bp), open reading frame (1,448 bp) and 3-regulatory flanking region (31 bp) by DNA walking. Based on this sequence, a 1.017 kb cDNA fragment encompassing the Open Reading Frame (ORF) that encodes for GPD1 was subsequently isolated by reverse transcription-polymerase chain reaction. Comparison of the GPD1 and its cDNA sequences demonstrated that the complete gene sequence encodes a polypeptide chain of 338 amino acids interrupted by 2 introns. Sequence comparison analysis of the 5 non-coding region with the 5 flanking sequences of other fungal GPD genes show several regions of similar sequence. The segments from positions -68 bp relative to the start codon is potentially a transcription start site and is mapped within the pyrimidine rich region. The presumptive TATA and CAAT boxes are mapped at -363 to -358 and -109 to -105 from the initiation of translation sites, respectively. The deduced protein product is 71 to 96% identical to glyceraldehyde-3-phosphate dehydrogenases of other filamentous fungi. Phylogenetic analysis based on deduced amino acid sequences shows that GPD1 of T. virens forms a cluster with filamentous ascomycetes. 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