Tag Archives: cDNA

Identification and heterologous expression of the cytochrome P450 oxidoreductase from the white-rot basidiomycete Coriolus versicolor.

Ichinose H, Wariishi H, Tanaka H.

Faculty of Agriculture, Kyushu University, Fukuoka, Japan.

Abstract

A cDNA encoding cytochrome P450 oxidoreductase (CPR) from the lignin-degrading basidiomycete Coriolus versicolor was identified using RT-PCR. The full-length cDNA consisted of 2,484 nucleotides with a poly(A) tail, and contained an open reading frame. The G+C content of the cDNA isolated was 60%. A deduced protein contained 730 amino acid residues with a calculated molecular weight of 80.7 kDa. The conserved amino acid residues involved in functional domains such as FAD-, FMN-, and NADPH-binding domains, were all found in the deduced protein. A phylogenetic analysis demonstrated that C. versicolor CPR is significantly similar to CPR of the basidiomycete Phanerochaete chrysosporium and that they share the same major branch in the fungal cluster. A recombinant CPR protein was expressed using a pET/ Escherichia coli system. The recombinant CPR protein migrated at 81 kDa on SDS polyacrylamide gel electrophoresis. It exhibited an NADPH-dependent cytochrome c reducing activity.

PMID: 12226721 [PubMed – indexed for MEDLINE]

http://www.ncbi.nlm.nih.gov/pubmed/12226721

CLONING OF SEQUENCES INDUCED AND SUPPRESSED BY ADMINISTRATION OF PSK, ANTITUMOR PROTEIN-BOUND POLYSACCHARIDE

Kunitaka Hirose, Michinori Hakozaki, Kenichi Matsunaga, Chikao Yoshikumi, Tetsuya Hotta, Masaaki Yanagisawa, Mikio Yamamoto, Hideya Endo.

To elucidate the effects of PSK, a protein-bound polysaccharide from Coriolus versicolor, on gene expression in tumor cells, we prepared cDNA clone libraries from PSKtreated and untreated cells of a rat ascites hepatoma line, AH66, which was previously shown to be susceptible to the antitumor action of this compound. Two PSK-induced and one suppressed cDNA clones were selected from these libraries by using a differential colony hybridization and RNA blot hybridization. PSK was thus shown to have a direct effect on the transcription and consequently on the translation of tumor cells.

Full article: http://mushroomstudies.co/wp-content/uploads/2010/09/Cloning-of-sequences-induced-and-suppressed-by-administration-of-PSK-antitumor-protein-bound-polysaccharide.pdf