Molecular cloning, nucleotide sequence and expression of the gene encoding prepro-polygalacturonaseII of Aspergillus niger.
PolygalacturonaseII of Aspergillus niger was fragmented using CNBr and the NH2-terminal fragment and another fragment were partially sequenced. The polygalacturonaseII (pgaII) gene was then isolated by using an oligonucleotide mixture based on the internal amino acid sequence as a probe. The nucleotide sequence of the pgaII structural gene was determined. It was found that polygalacturonaseII is synthesized as a precursor having an NH2-terminal prepro-sequence of 27 amino acids. The cloned gene was used to construct polygalacturonaseII over-producing A. niger strains. PolygalacturonaseII was isolated from one such strain and was determined to be correctly processed and to be fully active.
Bussink
H J
HJ
Department of Genetics, Agricultural University, Wageningen, The Netherlands.
Kester
H C
HC
Visser
J
J
D000595:Amino Acid Sequence; D001234:Aspergillus niger / Q000201:enzymology / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D004271:DNA, Fungal / Q000235:genetics; D004792:Enzyme Precursors / Q000235:genetics* / Q000302:isolation & purification; D015870:Gene Expression; D005800:Genes, Fungal; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D010957:Plasmids; D011096:Polygalacturonase / Q000235:genetics* / Q000302:isolation & purification; D015183:Restriction Mapping
D004271:DNA, Fungal; D004792:Enzyme Precursors; D015345:Oligonucleotide Probes; D011096:Polygalacturonase