| SDH-SV2C-L4融合蛋白的表达及山梨糖脱氢酶活性检测 |
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| 引用本文： | 韦娜,陈惠鹏,张惟材.SDH-SV2C-L4融合蛋白的表达及山梨糖脱氢酶活性检测[J].生物技术通讯,2011,22(3):354-357. |
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| 作者姓名： | 韦娜 陈惠鹏 张惟材 |
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| 作者单位： | 军事医学科学院,生物工程研究所,北京,100071 |
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| 摘 要： | 目的:构建SDH-SV2C-L4融合蛋白表达载体,在大肠杆菌中表达具有山梨糖脱氢酶(SDH)活性的融合蛋白。方法:将C亚型突触囊泡蛋白2大突环L4(SV2C-L4)基因与SDH基因以GGGS柔性接头连接,在大肠杆菌DH5α中表达;用NBT染色和DCIP脱色的方法检测融合蛋白的SDH活性。结果:DNA测序及SDS-PAGE结果显示构建了融合蛋白表达载体,并表达了SDH-SV2C-L4融合蛋白,相对分子质量约80×103;DCIP脱色及NBT染色均检测到融合蛋白的SDH活性。结论:与SV2C-L4融合的SDH仍具有活性,为下一步SV2C-L4活性检测方法的建立及SDH与SV2C-L4的其他相关研究奠定了基础。
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| 关 键 词： | 山梨糖脱氢酶 C亚型突触囊泡蛋白2大突环L4 融合蛋白 |
Expression of the Fusion Protein SDH-SV2C-L4 and Detection of its SDH Activity |
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| WEI Na,CHEN Hui-Peng,ZHANG Wei-Cai.Expression of the Fusion Protein SDH-SV2C-L4 and Detection of its SDH Activity[J].Letters in Biotechnology,2011,22(3):354-357. |
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| Authors: | WEI Na CHEN Hui-Peng ZHANG Wei-Cai |
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| Institution: | Beijing Institute of Biotechnology,Beijing 100071,China |
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| Abstract: | Objective：To construct a fusion expression vector for SDH-SV2C-L4 and express the fusion protein in E.coli with sorbose dehydrogenase（SDH） activity.Methods：The gene of SDH and synaptic vesicle protein 2C （SV2C）-L4 were linked with a GGGS linker and cloned into pET22b（＋）.The fusion vector was expressed in E.coli DH5α.SDH activity was detected by NBT staining and DCIP decolorizing.Results：DNA sequencing and SDS-PAGE analysis demonstrated that the fusion expression vector was successfully constructed,and the SDH-SV2C-L4 fusion protein was expressed.SDH activity was confirmed by both NBT staining and DCIP decolorizing.Conclusion：Recombinant SDH-SV2C-L4 fusion protein with SDH activity was obtained,which laid foundation for further study on SV2C-L4 and SDH. |
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| Keywords: | sorbose dehydrogenase synaptic vesicle protein 2C-L4 fusion protein |
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