Expression Plasmids for Use in Candida glabrata
نویسندگان
چکیده
We describe a series of CEN/ARS episomal plasmids containing different Candida glabrata promoters, allowing for a range of constitutive or regulated expression of proteins in C. glabrata. The set of promoters includes three constitutive promoters (EGD2pr, HHT2pr, PDC1pr), two macrophage/phagocytosis-induced promoters (ACO2pr, LYS21pr), and one nutritionally regulated promoter (MET3pr). Each promoter was cloned into two plasmid backbones that differ in their selectable marker, URA3, or the dominant-selectable NAT1 gene, which confers resistance to the drug nourseothricin. Expression from the 12 resulting plasmids was assessed using GFP as a reporter and flow cytometry or quantitative reverse-transcription polymerase chain reaction to assess expression levels. Together this set of plasmids expands the toolkit of expression vectors available for use with C. glabrata.
منابع مشابه
Evaluation of miR-146a expression level in macrophages exposed to Candida glabrata
Background and Purpose: MicroRNAs are small non-coding RNAs with 1924- nucleotides in length. Up- or downregulation of many miRNAs has been shown by stimulation of Toll-like receptors (TLRs) in the innate immune system. Up-regulation of miR-146a has been reported by both TLR and heat-killed Candida albicans. In this study, we aimed to evaluate the expression of miR-146a in cultured monocyte-...
متن کاملA system for gene cloning and manipulation in the yeast Candida glabrata.
The opportunistic pathogenic yeast, Candida (Torulopsis) glabrata, is an asexual imperfect fungus that exists largely as a haploid. Besides being a clinically important pathogen, this yeast also provides a model system for understanding basic biological mechanisms such as metal-activated metallothionein-encoding gene transcription. To facilitate molecular genetic studies in C. glabrata, we isol...
متن کاملIn vitro Antifungal Effect of Aqueous Garlic (Allium Sativum) Extract and its Combination with Fluconazole Against Five Common Clinical Candida Isolated from Candidiasis Lesions
Introduction: Increasing rate of candidiasis prevalence and consequently use of antifungal drugs as prophylactic and curative agents has led to the widespread emergence of resistant strains. Therefore this study was designed to evaluate the in vitro antifungal activity of an aqueous extract of garlic and the synergic effect of garlic extract with fluconazole against common clinical isolates of ...
متن کاملA Comparative Study on Level of erg11 Gene Expression in Non-albicans Candida Strains Isolated from Patients and Raw Milk
Nowadays, non-albicans Candida are common in human pathogens, and some of these cases were found in milk in the present study. Thanks to its various minerals and vitamins, milk is a good food source for humans and an ideal environment for the growth of microorganisms, including bacteria and fungi. Also, it is an important product in the agricultural sector. Materials and Methods: In the presen...
متن کاملEfficient homologous and illegitimate recombination in the opportunistic yeast pathogen Candida glabrata.
The opportunistic pathogen Candida glabrata causes significant disease in humans. To develop genetic tools to investigate the pathogenicity of this organism, we have constructed ura3 and his3 auxotrophic strains by deleting the relevant coding regions in a C. glabrata clinical isolate. Linearized plasmids carrying a Saccharomyces cerevisiae URA3 gene efficiently transformed the ura3 auxotroph t...
متن کامل