The Pichia pastoris HIS4 gene: nucleotide sequence, creation of a non-reverting his4 deletion mutant, and development of HIS4-based replicating and integrating plasmids

A clone of the Pichia pastoris HIS4 gene was obtained and its nucleotide sequence was determined. Based upon its deduced amino-acid sequence, the product of the P. pastoris HIS4 gene has the same structural organization as the Saccharomyces cerevisiae His4 protein and appears to encode a trifunction...

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Bibliographic Details
Published inCurrent genetics Vol. 265
Main Authors Crane, D.I, Gould, S.J. (Kennedy Krieger Research Inst., Baltimore (USA))
Format Journal Article
LanguageEnglish
Published 1995
Subjects
GENE
GENES
HIDROLASAS
HYDROLASE
MARCADORES GENETICOS
MARQUEUR GENETIQUE
MUTACION INDUCIDA
MUTATION PROVOQUEE
OXIDORREDUCTASAS
OXYDOREDUCTASE
PICHIA
PLASMIDE
PLASMIDIOS
SECUENCIA NUCLEOTIDICA
SEQUENCE NUCLEOTIDIQUE
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Summary:A clone of the Pichia pastoris HIS4 gene was obtained and its nucleotide sequence was determined. Based upon its deduced amino-acid sequence, the product of the P. pastoris HIS4 gene has the same structural organization as the Saccharomyces cerevisiae His4 protein and appears to encode a trifunctional enzyme catalyzing the second (phosphoribosyl-ATP pyrophosphohydrolase), third (phosphoribosyl-AMP cyclohydrolase), and tenth (histidinol dehydrogenase) steps in histidine biosynthesis. The chromosomal copy of the HIS4 gene was disrupted by homologous recombination, creating the strain SGY58. The his4Delta deletion mutation in this strain lacks the entire coding region of this gene and has a reversion rate that is undetectable. A set of complementary plasmids that carry the HIS4 gene was also developed. Among these are nine Escherichia coli-P. pastoris shuttle vectors that transform the his4 deletion mutant at high efficiency and an integration vector for creating site-specific alterations of the P. pastoris genome.
Bibliography:97B6050
F30
ISSN:0172-8083
1432-0983