Cloning and characterization of the abfB gene coding for the major α-l-arabinofuranosidase (ABF B) of Aspergillus niger

Michel J A Flipphi, Margreet van Heuvel, Peter van der Veen, Jaap Visser, Leo H. de Graaff

Research output: Article

58 Citations (Scopus)

Abstract

Based on amino-acid sequence data from Aspergillus niger α-l-arabinofuranosidase B (ABF B), and cyanogen bromide fragments derived thereof, deoxyoligonucleotide mixtures were designed to the 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.

Original languageEnglish
Pages (from-to)525-532
Number of pages8
JournalCurrent Genetics
Volume24
Issue number6
DOIs
Publication statusPublished - dec. 1993

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Aspergillus niger
Organism Cloning
Genes
Amino Acids
Cyanogen Bromide
Polymerase Chain Reaction
Beta vulgaris
Genomic Library
Uridine
Introns
Amino Acid Sequence
Plasmids
Carbon
Molecular Weight
DNA
Growth
Proteins

ASJC Scopus subject areas

  • Genetics

Cite this

Cloning and characterization of the abfB gene coding for the major α-l-arabinofuranosidase (ABF B) of Aspergillus niger. / Flipphi, Michel J A; van Heuvel, Margreet; van der Veen, Peter; Visser, Jaap; de Graaff, Leo H.

In: Current Genetics, Vol. 24, No. 6, 12.1993, p. 525-532.

Research output: Article

Flipphi, Michel J A ; van Heuvel, Margreet ; van der Veen, Peter ; Visser, Jaap ; de Graaff, Leo H. / Cloning and characterization of the abfB gene coding for the major α-l-arabinofuranosidase (ABF B) of Aspergillus niger. In: Current Genetics. 1993 ; Vol. 24, No. 6. pp. 525-532.
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