Isolation and characterization of glyceraldehyde-3-phosphate dehydrogenase gene of Trichoderma virens UKM1

S. S L Oh, Farah Diba Abu Bakar, A. M. Adnan, N. M. Mahadi, Osman Hassan, Abdul Munir Abd. Murad

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

In this study, the isolation and characterization of Trichoderma virens glyceraldehyde-3-phosphate dehydrogenase gene (GPD1) and its promoter is described. AcDNAclone 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. The sequence of this gene and its promoter can be used for the development of genetic tools in molecular studies of T. virens and in the expression of heterologous genes.

Original languageEnglish
Pages (from-to)194-203
Number of pages10
JournalBiotechnology
Volume8
Issue number2
DOIs
Publication statusPublished - 2009

Fingerprint

Glyceraldehyde-3-Phosphate Dehydrogenases
Trichoderma
Genes
5' Flanking Region
Nucleic Acid Regulatory Sequences
Open Reading Frames
Sequence Analysis
Complementary DNA
Fungal Genes
3' Flanking Region
Ascomycota
TATA Box
Initiator Codon
Transcription Initiation Site
Expressed Sequence Tags
Introns
Reverse Transcription
Walking
Amino Acid Sequence
Fungi

Keywords

  • Glyceraldehyde-3-phosphate dehydrogenase
  • Promoter
  • Trichoderma virens

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Isolation and characterization of glyceraldehyde-3-phosphate dehydrogenase gene of Trichoderma virens UKM1. / Oh, S. S L; Abu Bakar, Farah Diba; Adnan, A. M.; Mahadi, N. M.; Hassan, Osman; Abd. Murad, Abdul Munir.

In: Biotechnology, Vol. 8, No. 2, 2009, p. 194-203.

Research output: Contribution to journalArticle

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