Yeast thioredoxin genes

Based on the conserved protein sequence of thioredoxins from yeast and other organisms, two primers were synthesized for polymerase chain reaction of yeast genomic DNA. A 34-base pair (bp) sequence around the active site of yeast thioredoxin was obtained from the polymerase chain reaction product. T...

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Bibliographic Details
Published in:The Journal of biological chemistry 1991-01, Vol.266 (3), p.1692-1696
Main Author: Gan, Z.R. (Merck Sharp and Dohme Research Laboratory, West Point, PA)
Format: Article
Language:English
Subjects:
ADN
Amino Acid Sequence
Base Sequence
BIOCHIMIE
Biological and medical sciences
BIOQUIMICA
Blotting, Southern
CLONACION
CLONAGE
cloning
Cloning, Molecular
DNA, Fungal - genetics
Fundamental and applied biological sciences. Psychology
Fungal Proteins - genetics
genes
Genes, Fungal
Genes. Genome
Molecular and cellular biology
Molecular genetics
Molecular Sequence Data
NUCLEOTIDE
nucleotide sequence
NUCLEOTIDOS
Polymerase Chain Reaction
prediction
PROTEINE SOUFREE
SACCHAROMYCES CEREVISIAE
Saccharomyces cerevisiae - genetics
Species Specificity
SULFOPROTEINAS
thioredoxin I
thioredoxin II
Thioredoxins - genetics
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Summary:Based on the conserved protein sequence of thioredoxins from yeast and other organisms, two primers were synthesized for polymerase chain reaction of yeast genomic DNA. A 34-base pair (bp) sequence around the active site of yeast thioredoxin was obtained from the polymerase chain reaction product. This specific sequence was used as a probe in Southern blot analysis of total yeast genomic DNA digested with various restriction enzymes. Under conditions of high stringency, more than one DNA species hybridized with the probe, suggesting that more than one gene encodes yeast thioredoxin. The probe was used to screen a yeast genomic library. Two Sau3A1 fragments, 825 and 2045 bp, respectively, from two different clones were cloned into pUC13. Sequence analysis of these fragments gave two different open reading frames without introns. The 825-bp Sau3A1 fragment encodes a 103-amino acid residue protein named thioredoxin I. The 2045-bp Sau3A1 fragment contains a sequence encoding thioredoxin II which has 102 amino acid residues. This is the first report of the cloning and sequencing of eukaryotic thioredoxin genes from any source. Both yeast thioredoxins contain a dithiol active site sequence, Cys-Gly-Pro-Cys. Thioredoxins I and II show 78% amino acid sequence identity. They display more amino acid sequence similarity with mammalian thioredoxin than with Escherichia coli and plant chloroplast thioredoxins
ISSN:0021-9258
1083-351X
DOI:10.1016/S0021-9258(18)52350-4