Cloning and characterization of a thioredoxin gene, CpTrx1, from the chestnut blight fungus Cryphonectria parasitica.
A differential display for the expression profiles of wild-type Cryphonectria parasitica and its virally-infected isogenic hypovirulent strain revealed several transcripts of interest, which evidenced significant matches with fungal genes of known function. Among which, we have further analyzed an amplified PCR product with significant sequence similarity to the known fungal stress-responsive thioredoxin gene from Neurospora crassa. The product of the cloned thioredoxin gene, CpTrx1, consists of 117 amino acids, with a predicted molecular mass of 13.0 kDa and a pI of 5.4. Sequence comparisons demonstrated that the deduced protein sequence of the CpTrx1 gene evidenced a high degree of homology to all known thioredoxins, with the highest degree of homology with trx1, a thioredoxin gene from Saccharomyces cerevisiae, and evidenced a preservation of the conserved hall mark-residues (Trp-Cys-Gly-Pro-Cys) at the active site of thioredoxin. The E. coli-generated CpTRX1 manifested thioredoxin activity, according to the insulin reduction assay, which indicates that the cloned gene does indeed encode for the C. parasitica thioredoxin.
Kim
Ji-Hye
JH
Basic Science Research Institute, Institute for Molecular Biology and Genetics, Chonbuk National University, Jeonju 561-756, Republic of Korea.
Kim
Dae-Hyuk
DH
D000595:Amino Acid Sequence; D001203:Ascomycota / Q000145:classification / Q000235:genetics*; D015152:Blotting, Northern; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal / Q000187:drug effects; D008969:Molecular Sequence Data; D010802:Phylogeny; D011994:Recombinant Proteins / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013879:Thioredoxins / Q000235:genetics* / Q000378:metabolism; D024483:Vitamin K 3 / Q000494:pharmacology; D014815:Vitamins / Q000494:pharmacology
D005656:Fungal Proteins; D011994:Recombinant Proteins; D014815:Vitamins; D013879:Thioredoxins; D024483:Vitamin K 3