Cloning and Characterization of the abfB Gene Coding for the Major alpha-L-arabinofuranosidase (ABF B) of Aspergillus Niger

Curr Genet. 1993 Dec;24(6):525-32. doi: 10.1007/BF00351717.

Abstract

Based on amino-acid sequence data from Aspergillus niger alpha-L-arabinofuranosidase B (ABF B), and cyanogen bromide fragments derived thereof, deoxyoligonucleotide mixtures were designed to be employed as primers in a polymerase chain reaction (PCR) on A. niger genomic DNA. This resulted in amplification of three related PCR products. The abfB gene encoding ABF B was isolated from a genomic library using such an amplification product as a probe. A 5.1-kb BamHI fragment was subcloned to result in plasmid pIM991. Upon introduction by co-transformation into both A. niger and A. nidulans uridine auxotrophic strains, pIM991 was shown to contain the functional gene since prototrophic transformants overproduced ABF B upon growth on the inducing carbon source sugar beet pulp. A plate assay was developed enabling quick selection of ABF B-overproducing transformants. The sequence of a 4122-bp long BamHI/SstI fragment was determined. The abfB gene does not contain introns and codes for a protein of 499 amino acids. The mature ABF B, 481 amino acids in length, has a deduced molecular weight of 50.7 kDa. A. niger abfB is the first eukaryotic gene encoding an ABF to be characterized.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Aspergillus nidulans / enzymology
  • Aspergillus nidulans / genetics
  • Aspergillus niger / enzymology
  • Aspergillus niger / genetics*
  • Base Sequence
  • Cloning, Molecular
  • DNA Primers
  • DNA, Fungal / metabolism
  • Gene Expression
  • Genes, Fungal*
  • Glycoside Hydrolases / biosynthesis
  • Glycoside Hydrolases / chemistry
  • Glycoside Hydrolases / genetics*
  • Molecular Sequence Data
  • Peptide Fragments / chemistry
  • Peptide Mapping
  • Plasmids
  • Polymerase Chain Reaction
  • Restriction Mapping

Substances

  • DNA Primers
  • DNA, Fungal
  • Peptide Fragments
  • Glycoside Hydrolases
  • alpha-N-arabinofuranosidase

Associated data

  • GENBANK/L23502
  • GENBANK/X70022
  • GENBANK/X70023
  • GENBANK/X70024
  • GENBANK/X70025
  • GENBANK/X70026
  • GENBANK/X70027
  • GENBANK/X70028
  • GENBANK/X71679
  • GENBANK/X71680
  • GENBANK/X74777