利用RNA干扰沉默马尔尼菲青霉菌聚酮合酶基因的相关研究

RNA Interference in Penicillium marneffei Demonstrates a Role for cps Gene in Pigmentation Biosynthesis

在实验室前期建立的马尔尼菲青霉菌cDNA文库中筛到了1个马尔尼菲青霉菌聚酮合酶基因cps(Cit-rinin Polyketide Synthase),为了研究这个cps基因的功能,构建了RNAi表达盒,表达盒内具有目的基因编码序列422bp的反向重复序列并被gus基因隔开.整个干扰载体借助根癌农杆菌体系成功转化马尔尼菲青霉菌,并受木糖的诱导启动子xylP调控RNA干扰.研究发现cps基因沉默的转化子产生与母代红色菌株不同的白色菌株,类似基因敲除的表型,表明cps基因与该青霉菌色素的形成直接相关.

To explore the gene function of cps gene,which encodes polyketide synthase and screened from cDNA library of Penicillium marneffei,a RNAi expression cassette was designed with inverted repeats of coding sequence of the targeted gene separated by a segment of gus gene.Constructs were transferred by Agrobacterium tumefaciesns-mediated transformation and under the control of promoter xylP,which promoter can be induced by xylose.The cps-silenced transformants demonstrated a clear white phenotype,instead of red ...