Molecular Cloning of the Imidazoleglycerolphosphate Dehydratase Gene of Trichoderma Harzianum by Genetic Complementation in Saccharomyces Cerevisiae Using a Direct Expression Vector

Overview

Journal Mol Gen Genet

Specialties Genetics
Molecular Biology

Date 1992 Sep 1

PMID 1406594

Citations 7

Authors

G H Goldman

J Demolder

S Dewaele

A Herrera-Estrella

R A Geremia

M Van Montagu

R Contreras

Affiliations

Soon will be listed here.

Abstract

The Trichoderma harzianum imidazoleglycerolphosphate dehydratase gene (igh) has been isolated by complementation of a Saccharomyces cerevisiae his3 mutant using a direct expression vector. This Escherichia coli-yeast shuttle vector was developed to allow efficient cloning and expression of cDNA libraries. The cDNA is 627 nucleotides long and codes for a protein of 209 amino acids with an apparent molecular mass of 22,466 daltons. The predicted protein sequence showed 63.6%, 58.7%, and 38.4% identity respectively to the corresponding enzymes from S. cerevisiae, Pichia pastoris and E. coli. Northern analysis showed that the expression of the igh gene in T. harzianum is not inhibited by external histidine and the level of igh mRNA was about threefold higher in cells starved of histidine.

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