Cloning and sequence analysis of the Pichia pastoris TRP1, IPP1 and HIS3 genes.
The Pichia pastoris TRP1 and HIS3 genes were cloned by complementation of the Saccharomyces cerevisiae trip1 and his3 mutants, respectively, and their nucleotide sequence was determined. The P. pastoris TRP1 gene includes an open reading frame (ORF) of 714 nucleotides corresponding to a polypeptide of 237 amino acids whose sequence shares about 40% identity with that of TRP1 encoding proteins in other yeast species. DNA sequencing showed that an ORF of 858 nucleotides, encoding a protein of 285 amino acids with high homology to inorganic pyrophosphatases (IPP1), is located downstream of the P. pastoris TRP1 gene. Both genes converge in this chromosomal region, showing a genetic organization analogous to that found in the Kluyveromyces lactis genome. The P. pastoris HIS3 gene possesses an ORF of 675 nucleotides, encoding a polypeptide of 224 amino acids which shows 74.1% identity to the homologous S. cerevisiae protein. The hexameric consensus GCN4 binding sequence (TGACTC), characteristic of many amino acid biosynthetic genes, is present in the promoter region.
Cosano
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Departamento de Microbiología II, Facultad de Farmacia, Universidad Complutense, Madrid, Spain.
Alvarez
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P
Molina
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Nombela
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C
D019747:Aldose-Ketose Isomerases; D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics*; D005800:Genes, Fungal; D006836:Hydro-Lyases / Q000737:chemistry / Q000235:genetics*; D043564:Inorganic Pyrophosphatase; D008969:Molecular Sequence Data; D010843:Pichia / Q000235:genetics*; D011755:Pyrophosphatases / Q000737:chemistry / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D015003:Yeasts / Q000235:genetics
D005656:Fungal Proteins; D029701:Saccharomyces cerevisiae Proteins; D011755:Pyrophosphatases; D043564:Inorganic Pyrophosphatase; D006836:Hydro-Lyases; C021434:imidazoleglycerolphosphate dehydratase; D019747:Aldose-Ketose Isomerases; C490353:TRP1 protein, S cerevisiae