摘要
目的获得一种适于丝状真菌基因研究用的、由根癌农杆菌介导的RNA干扰方法。方法采用基因重组及菌株干扰体系筛选的方法。结果获得适于根癌农杆菌转染的重组载体PCB309-pfgrt,并将其成功用于对孢子丝菌双组份信号组氨酸蛋白激酶DRK1基因的干扰中。结论该方法优化了根癌农杆菌的转化体系,解决了丝状真菌RNA干扰载体构建困难及干扰效率低下的难题,在丝状真菌基因功能及遗传转化研究方面具有广泛的应用前景。
Objective To obtain a RNA-interference method mediated by Agrobacterium tumefaciens applicable for gene research on filamentous fungi.Methods Gene recombination and interference system selection were used. Results The recombinant vector PCB309-pfgrt was obtained and used in interfering the expression level of two-component DRK1 gene successfully.Conclusions This method optimized the transformation system,solving the problem of high efficient RNA interference vector construction,which has wide application prospect in genetic research in filamentous fungi.
出处
《中国真菌学杂志》
CSCD
2015年第3期163-167,共5页
Chinese Journal of Mycology
基金
国家自然科学基金(81000699)
深圳市科创委(JCYJ20140414092023239)
关键词
重组载体
RNA干扰
丝状真菌
根癌农杆菌
recombinant vector
RNA-interference
filamentous fungi
Agrobacterium tumefaciens
