Menino João F, Almeida Agostinho J, Rodrigues Fernando
Life and Health Sciences Research Institute (ICVS), School of Health Sciences, University of Minho, Braga, Portugal.
Methods Mol Biol. 2012;845:187-98. doi: 10.1007/978-1-61779-539-8_12.
Paracoccidioides brasiliensis is a thermal dimorphic fungus which in the host environment exhibits a multinucleated and multibudding yeast form. The cellular and molecular mechanisms underlying these phenotypes remain to be clarified, mostly due to the absence of efficient classical genetic and molecular techniques. Here we describe a method for gene expression knockdown in P. brasiliensis by antisense RNA (aRNA) technology taking advantage of an Agrobacterium tumefaciens-mediated transformation (ATMT) system. Together, these techniques represent a reliable toolbox that can be employed for functional genetic analysis of putative virulence factors and morphogenic regulators, aiming to the identification of new potential drug targets.
巴西副球孢子菌是一种温度双相真菌，在宿主环境中呈现多核和多芽酵母形态。这些表型背后的细胞和分子机制仍有待阐明，主要是由于缺乏有效的经典遗传和分子技术。在此，我们描述了一种利用根癌农杆菌介导的转化（ATMT）系统，通过反义RNA（aRNA）技术在巴西副球孢子菌中敲低基因表达的方法。这些技术共同构成了一个可靠的工具包，可用于对假定的毒力因子和形态发生调节因子进行功能基因分析，旨在识别新的潜在药物靶点。
