【摘要】 从本实验室早期建立的马尔尼菲青霉cDNA文库中,筛选到了一个编码环腺苷酸依赖性蛋白激酶(cAMP dependent protein kinase)基因cdpk.为了研究这个基因的功能,构建了相应的RNA干扰表达盒(利用gus基因将cdpk基因313 bp的反向重复序列片段隔开,并由受木糖诱导的xylP启动子来调控RNA干扰),通过根癌农杆菌介导转化到马尔尼菲青霉中.对转化子的研究发现,cdpk基因的沉默对菌体细胞的形态、孢子的萌发过程无明显影响,但是却减弱了菌落的生长速度、降低了红色色素的产量、并使马尔尼菲青霉几乎丧失了产孢功能.这暗示着cdpk基因对该青霉孢子的形成是必需的.更多还原

【Abstract】 In order to study the function of cdpk gene,which encodes cAMP dependent protein kinase and screened from cDNA library of Penicillium marneffei,a RNAi expression cassette was designed.This cassette contained inverted repeats of 313 bp coding sequence of the targeted gene separated by a segment of gus gene,and under the control of xylP promoter induced by xylose to regulate the RNAi.Via the Agrobacterium tumefaciesns-mediated transformation,the RNAi expression cassette was introduced into Penicillium marneffei.The cdpk-silenced transformants displayed no visible change in cell morphogenesis and the conidia germination,but reduced in growth rates and pigment production.The most severe change of phenotype after cdpk silence was almost no conidia formed during asexual development.These evidences suggest that cdpk gene is essential for sporulation of Penicillium marneffei.更多还原