[Characterization of 60 kDa heat shock protein [HSP60] gene in pathogenic dermatophyte Microsporom canis]

The stress-inducible Heat Shock Proteins [i.e. HSP60] constitute one of the highly conserved protein and gene families. As one of the molecular chaperone proteins, they play essential roles in protein metabolism and protein translocation under both stress and non-stress conditions. In the present study, we try to characterize the 60 kDa Heat Shock Protein [HSP60] gene in dermatophyte pathogen Microsporom Canis [M. canis]. This dermatophyte is one of the most important causative agents of dermatophytosis in human and animals. Some properties of M. canis have been investigated in molecular level; however, no information is available regarding the HSP60 in this dermatophyte. In the present study, we try to characterize the 60 kDa Heat Shock Protein [HSP60] gene in dermatophyte pathogen Microsporom Canis [M. canis]. M. canis was obtained from patients with dermatophytosis and cultured in appropriate conditions. High molecular weight DNA was isolated from obtained mycelial mass by standard methods. Pairs of 21 nt primers were designed from highly conserved regions of the HSP60 genes in other eukaryotic cells. Mentioned primers were utilized in PCR using isolated genomic DNA template of M. canis. Predicted molecules have been amplified and were submitted for sequencing. By the time 1550 nucleotides of this gene are sequenced and analysed, that encoding a 497 amino acids protein. In the present study, we report the identification and molecular characterization of a M. canis gene encoding a protein belongs to the 60 kDa Heat Shock Protein family which will here be referred to as McHSP60. Analysis of the amino acid sequence of this gene revealed a considerable identity with other eukaryotic HSP60 such as those of C.immitis [97%], Aspergillus fumigatus [92%] and S.cerevisiae [74%]. Investigation of amino acid composition in HSP60 revealed Alanine, Glutamic acid and Glysine as the most common amino acids in this protein. The amino acid composition of McHSP60 indicates the amount of Cysteine and Tryptophan are poor

Study of griseofulvin effects on the expression of ATPase-subunit G gene in dermatophyte pathogen Trichophyton rubrum

Trichophyton rubrum is one of the anthropophilic dermatophytes with worldwide distribution. This fungus is a common causative agents of Tinea cruris, Tinea corporis, Tinea pedis and tinea manuum. Several properties of this fungus have been studied so far; however few investigations were carried out in the field of molecular biology of this microorganism. The main goal of this survey was the evaluation of ATPase-subunit G gene expression of this fungus due to different amounts of griseofulvin. Serial dilutions of griseofulvin with fungal growth were prepared and compared with control. In this study, the control and griseofulvin-treated samples were micro scopically investigated and the RNAs were then extracted. Consequently RT-PCR was performed simultaneously in order to evaluate griseofulvin influence on ATPase-subunit G gene expression. It was indicated that the rate of shortened and twisted mycelia in 10 micro g/ml of griseofulvin was much higher than micro g/ml. Meanwhile, in micro g/ml of this drug there is an obvious up-regulation in ATPase-subunit G gene in comparison with 10 micro g/ml. It can be concluded that in certain amounts of griseofulvin, a significant increment in ATPase-subunit G gene expression as well as mycelial abnormalities could be occurred