关键词: 申克孢子丝菌, STE20, RNA干扰, 载体构建

Abstract:

Objective This study aimed to construct a universal RNA interference vector of fungi, and to be widely used in the research of gene interference of various fungi. Methods Using the plant double expression vector, multi-cloning site (MCS) of pBlueScriptⅡ was cut down to construct our interference vector. To make the vector suitable for Agrobacterium transformation, we added t-DNA boundary repeats. In order to facilitate screening after transformation, hygromycin resistance genes were constructed into interfering vectors. In order to expand the interference range, we amplified the universal promoter ptrpc, spacer sequence and terminator Ttrpc in psilent-1 plasmid, and introduced hygromycin resistance gene to construct the vector. Results The vector PCB309-PSUST constructed in this experiment and the optimized reaction system were successfully used to effectively interfere with the STE20 gene of Sporothrix, and the operation was simple, the transformation efficiency was high, and the transformants were stable. Conclusion The constructed vector is suitable for Agrobacterium tumefaciens mediation and can be used for RNA interference research of filamentous fungal genes.

Key words: Sporothrix schenckii, STE20, RNA interference, vector construction