CAJ | 학술논문

目的：通过过表达小分子RNA和抑制小分子RNA功能，研究小分子RNA对里氏木霉QM9414纤维素酶表达的影响。方法：利用含里氏木霉强启动子Ppdc的质粒p-Ppdc'-Tcbh1，分别构建过表达载体和Tough Decoy（TuD）序列，对里氏木霉QM9414的3个小分子RNA（milR5、milR7和milR10）进行过表达和抑制，过表达和抑制后的小分子RNA表达量通过荧光定量PCR检测，最终测定转化菌株的滤纸酶活，研究过表达和抑制小分子RNA后对纤维素酶表达的影响。结果：过表达载体能提高小分子RNA的表达量，过表达重组菌株OE-milR7的滤纸酶活明显提高；TuD序列表达载体可抑制小分子RNA的功能，抑制小分子RNA的重组菌株TuD7酶活略有下降。结论：milR7可能参与纤维素酶表达调控，为对里氏木霉的产纤维素酶能力进行改造提供了新的靶点。
목적：통과과표체소분자RNA화억제소분자RNA공능，연구소분자RNA대리씨목매QM9414섬유소매표체적영향。방법：이용함리씨목매강계동자Ppdc적질립p-Ppdc'-Tcbh1，분별구건과표체재체화Tough Decoy（TuD）서렬，대리씨목매QM9414적3개소분자RNA（milR5、milR7화milR10）진행과표체화억제，과표체화억제후적소분자RNA표체량통과형광정량PCR검측，최종측정전화균주적려지매활，연구과표체화억제소분자RNA후대섬유소매표체적영향。결과：과표체재체능제고소분자RNA적표체량，과표체중조균주OE-milR7적려지매활명현제고；TuD서렬표체재체가억제소분자RNA적공능，억제소분자RNA적중조균주TuD7매활략유하강。결론：milR7가능삼여섬유소매표체조공，위대리씨목매적산섬유소매능력진행개조제공료신적파점。
Objective: By over-expressing the small RNAs and inhibiting the small RNAs function, investigating the regulation function of small non-coding RNAs on cellulase expression in Trichoderma reesei QM9414. Meth?ods: For studying the effect of over-expressing 3 small RNAs(milR5, milR7 and milR10) and inhibiting its func?tion on cellulase expression in T.reesei QM9414, the plasmid p-Ppdc'-Tcbh1 which contained a T.reesei strong pro?moter Ppdc was used to constructed over-expression cassettes and Tough Decoy(TuD) technology. The expression quantity of small RNAs was tested by real-time PCR, and the the cellulase expression was tested by filter paper activity(FPA). Results: The expression quantity of small RNAs showed that over-expression cassette enhanced the expression of the small RNAs, and the TuD technology inhibited the function of the small RNAs, respectively. The FPA showed that the recombinant strains OE-milR7 improved FPA distinctly, and TuD7 decreased FPA. Con?clusion: One of the small RNAs, milR7, might participate in the regulation of cellulase expression, which demon?strates a new strategy for increasing cellulase activities in T.reesei.