Cloning and sequence analysis of the glyceraldehyde-3-phosphate dehydrogenase gene from the zygomycetes fungus Rhizomucor miehei

Mária Vastag, Zsolt Kasza, Klára Ács, T. Papp, Helmut Schwab, C. Vágvölgyi

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Rhizomucor miehei is important from a biotechnological aspect in consequence of its content of aspartic proteinase, which has high milk-clotting activity. A genomic library of R. miehei NRRL 5901 has been constructed in a phage (Lambda Fix II) vector. The glyceraldehyde-3-phosphate dehydrogenase (gpd) gene was isolated from this library by hybridization of the recombinant phage clones with a gpd-specific gene probe generated by polymerase chain reaction. The complete nucleotide sequence encodes a putative polypeptide chain of 336 amino acids interrupted by 5 introns. The predicted amino acid sequence of this gene shows a high degree of sequence similarity to the glyceraldehyde-3-phosphate dehydrogenase proteins from yeast and filamentous fungi. The promoter region, containing a consensus TATA box, and 246-bp downstream from the putative stop codon were also determined. The possibility of using the gpd promoter in the construction of new transformation vectors is discussed.

Original languageEnglish
Pages (from-to)111-119
Number of pages9
JournalAntonie van Leeuwenhoek, International Journal of General and Molecular Microbiology
Volume86
Issue number2
DOIs
Publication statusPublished - Aug 2004

Fingerprint

Rhizomucor
Glyceraldehyde-3-Phosphate Dehydrogenases
Sequence Analysis
Organism Cloning
Fungi
Genes
Aspartic Acid Proteases
Bacteriophage lambda
TATA Box
Fungal Proteins
Genomic Library
Terminator Codon
Genetic Promoter Regions
Bacteriophages
Introns
Libraries
Amino Acid Sequence
Milk
Clone Cells
Amino Acids

Keywords

  • Cloning
  • Glyceraldehyde-3-phosphate dehydrogenase gene
  • Polymerase chain reaction
  • Rhizomucor miehei

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

Cite this

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AU - Vastag, Mária

AU - Kasza, Zsolt

AU - Ács, Klára

AU - Papp, T.

AU - Schwab, Helmut

AU - Vágvölgyi, C.

PY - 2004/8

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N2 - Rhizomucor miehei is important from a biotechnological aspect in consequence of its content of aspartic proteinase, which has high milk-clotting activity. A genomic library of R. miehei NRRL 5901 has been constructed in a phage (Lambda Fix II) vector. The glyceraldehyde-3-phosphate dehydrogenase (gpd) gene was isolated from this library by hybridization of the recombinant phage clones with a gpd-specific gene probe generated by polymerase chain reaction. The complete nucleotide sequence encodes a putative polypeptide chain of 336 amino acids interrupted by 5 introns. The predicted amino acid sequence of this gene shows a high degree of sequence similarity to the glyceraldehyde-3-phosphate dehydrogenase proteins from yeast and filamentous fungi. The promoter region, containing a consensus TATA box, and 246-bp downstream from the putative stop codon were also determined. The possibility of using the gpd promoter in the construction of new transformation vectors is discussed.

AB - Rhizomucor miehei is important from a biotechnological aspect in consequence of its content of aspartic proteinase, which has high milk-clotting activity. A genomic library of R. miehei NRRL 5901 has been constructed in a phage (Lambda Fix II) vector. The glyceraldehyde-3-phosphate dehydrogenase (gpd) gene was isolated from this library by hybridization of the recombinant phage clones with a gpd-specific gene probe generated by polymerase chain reaction. The complete nucleotide sequence encodes a putative polypeptide chain of 336 amino acids interrupted by 5 introns. The predicted amino acid sequence of this gene shows a high degree of sequence similarity to the glyceraldehyde-3-phosphate dehydrogenase proteins from yeast and filamentous fungi. The promoter region, containing a consensus TATA box, and 246-bp downstream from the putative stop codon were also determined. The possibility of using the gpd promoter in the construction of new transformation vectors is discussed.

KW - Cloning

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KW - Polymerase chain reaction

KW - Rhizomucor miehei

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