耐热木聚糖酶xynB_(64)和红色荧光蛋白Dsred2的融合表达及其应用

Co-expression and Application of Thermostable Xylanase xynB_(64) and Red Fluorescent Protein Dsred2

[目的]耐热木聚糖酶xynB64的表达研究,探讨利用红色荧光蛋白Dsred2标签简便快捷地检测包涵体复性的效率。[方法]构建重组质粒pET42a-xynB64和pET42a-xynB64-Dsred2,利用宿主菌Rosetta(DE3)进行表达,DNS法测定酶活,并检测其荧光强度和尿素复性的包涵体。[结果]红色荧光蛋白Dsred2促进了目的蛋白可溶性表达,重溶包涵体经激发后发射光波长在583 nm,光强度与可溶性大致成正比。[结论]研究表明红色荧光蛋白Dsred2在包涵体复性中可作为报告蛋白,该结果为工业化复性包涵体提供了检测依据。

[Objective] To improve the expression of thermostable xylanase xyB64 and easily detect the refolding rate of inclusion body by using co-expressing red fluorescent protein Dsred2.[Method] The recombinant plasmids pET42a-xynB64 and pET42a-xynB64-Dsred2 were transformed into the host Rosetta（DE3） respectively,the enzyme activity was determined via DNS,the fluorescence intensity and the inclusion body renatured by urea were detected.[Result] Red fluorescent protein promoted the soluble expression of target protein,and its refolding production emission wavelength was 583 nm when excited.Fluorescence intensity was roughly coincident with dissolubility.[Conclusion] The red fluorescent protein Dsred 2 could be the report protein during the renaturing of inclusion body,and the research provided the basis for industrial protein production.