Cloning, sequence and structure of a gene encoding an antifungal glucan 1,3-beta-glucosidase from Trichoderma atroviride (T. harzianum)

Gene. 2001 Oct 17;277(1-2):199-208. doi: 10.1016/s0378-1119(01)00681-3.

Abstract

A gene (gluc78) encoding an antifungal glucan 1,3-beta-glucosidase was cloned from strain P1 of the biocontrol fungus Trichoderma atroviride (formerly T. harzianum). A putative regulatory sequence upstream from the coding region was cloned using single-strand extension from a primer in the known portion of the gene, circularized with T4 ligase, and then reamplified with PCR to generate double-stranded DNA. The entire genomic DNA sequence consisted of 3440 bp, with 559 and 579 bp, respectively, in 5' and 3' untranslated regions. The transcription unit contains a single intron, positioned in the 5' untranslated region. The gene encodes for a protein of 770 aa, including a 40 aa signal peptide. Symmetry between the first and second halves of the mature protein was found. The gene is present as a single copy in T. atroviride and a similar gene also is present in T. harzianum and T. virens. The encoded protein has similarity to a small group of sequences from filamentous fungi and no significant similarity to 1,3-beta-glucanases or glucosidases from other organisms. Northern analysis indicates that the gene is repressed in the presence of 3% glucose and expressed in media containing 0.1% of the sugar. Laminarin (0.1%) enhances expression after 18 h and other polymers such as scleroglucan and pustulan may enhance expression after 40 h of growth.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • 5' Flanking Region / genetics
  • Amino Acid Sequence
  • Blotting, Northern
  • Blotting, Southern
  • Cloning, Molecular
  • DNA, Fungal / chemistry
  • DNA, Fungal / genetics
  • Exons
  • Fungi / enzymology
  • Gene Expression Regulation, Enzymologic
  • Gene Expression Regulation, Fungal
  • Genes / genetics*
  • Glucan 1,3-beta-Glucosidase
  • Introns
  • Molecular Sequence Data
  • RNA, Fungal / genetics
  • RNA, Fungal / metabolism
  • Regulatory Sequences, Nucleic Acid / genetics
  • Sequence Alignment
  • Sequence Analysis, DNA
  • Sequence Homology, Amino Acid
  • Transcription Initiation Site
  • Trichoderma / enzymology
  • Trichoderma / genetics*
  • beta-Glucosidase / genetics*

Substances

  • DNA, Fungal
  • RNA, Fungal
  • beta-Glucosidase
  • Glucan 1,3-beta-Glucosidase

Associated data

  • GENBANK/AF253421