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 |
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引用本文: | 白羽,申宁. 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[J]. 安徽农业科学, 2012, 0(7): 3891-3893 |
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作者姓名: | 白羽 申宁 |
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作者单位: | 南京工业大学,江苏南京,211800 |
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摘 要: | [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。
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关 键 词: | 耐热木聚糖酶xynB64 红色荧光蛋白Dsred2 原核表达 包涵体 |
Co-expression and Application of Thermostable Xylanase xynB64 and Red Fluorescent Protein Dsred2 |
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Affiliation: | BAI Yu et al(Nanjing University of Technology,Nanjing,Jiangsu 211800) |
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Abstract: | [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. |
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Keywords: | Thermostable xylanase xyB64 Red fluorescent protein Dsred2 Prokaryotic expression Inclusion body |
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