| 钝顶节旋藻(Arthrospira platensis)2-DE分析蛋白制备方法改进及试用于螺旋手性差异蛋白研究 |
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| 引用本文： | 王景梅,汪志平,于金鑫,刘新颖,邵 斌,蓝瑾瑾,马丽芳,陈子元.钝顶节旋藻(Arthrospira platensis)2-DE分析蛋白制备方法改进及试用于螺旋手性差异蛋白研究[J].海洋与湖沼,2013,44(1):141-147. |
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| 作者姓名： | 王景梅 汪志平 于金鑫 刘新颖 邵 斌 蓝瑾瑾 马丽芳 陈子元 |
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| 作者单位： | 浙江大学原子核农业科学研究所农业部核农学重点开放实验室 杭州 310029 |
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| 基金项目： | 国家自然科学基金项目, 30771669 号, 10975118 号; 国家公益性行业(农业)科研专项, 201107003 号; 浙江省院士基金, J20080388号, J20110445 号。 |
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| 摘 要： | 采用冻融破壁和蛋白质分步提取等技术,对钝顶节旋藻2-DE分析蛋白制备方法作了改进,并试用于螺旋手性差异表达蛋白研究,结果表明:(1)钝顶节旋藻细胞反复冻融5次完全破碎,用Tris-HC1提取液提取3次可提得近87％的水溶性蛋白,再用SDS(十二烷基磺酸钠)提取液可从沉淀中提得水不溶性蛋白;(2)将上述水溶性和水不溶性蛋白分别用TCA/丙酮法纯化后作双向电泳(2-DE)分析,可辨蛋白点分别达500和760多个,而两者间的匹配率仅7％,说明分离度好且质量高；(3)利用此法获得了13个与钝顶节旋藻螺旋手性相关的候选蛋白,其中5个为首次发现,候选蛋白功能涉及光合作用、能量代谢、物质外排和环境适应等；(4)本文所建的“冻融破壁分步提纯法”与现有节旋藻2-DE蛋白制备方法相比,具有操作简便、仪器设备简单、成本低廉及信息量更丰富等优点.
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| 关 键 词： | 钝顶节旋藻 蛋白质 制备方法 双向电泳 螺旋手性 |
| 收稿时间： | 2011/9/15 0:00:00 |
| 修稿时间： | 4/6/2012 12:00:00 AM |
AN IMPROVED METHOD OF 2-DE ANALYSIS PROTEIN PREPARATION AND USING FOR INVESTIGATION OF HELICAL-HANDED DIFFERENTIAL PROTEINS IN ARTHROSPIRA PLATENSIS |
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| WANG Jing-Mei,WANG Zhi-Ping,YU Jin-Xin,LIU Xin-Ying,SHAO Bin,LAN Jin-Jin,MA Li-Fang and CHEN Zi-Yuan.AN IMPROVED METHOD OF 2-DE ANALYSIS PROTEIN PREPARATION AND USING FOR INVESTIGATION OF HELICAL-HANDED DIFFERENTIAL PROTEINS IN ARTHROSPIRA PLATENSIS[J].Oceanologia Et Limnologia Sinica,2013,44(1):141-147. |
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| Authors: | WANG Jing-Mei WANG Zhi-Ping YU Jin-Xin LIU Xin-Ying SHAO Bin LAN Jin-Jin MA Li-Fang and CHEN Zi-Yuan |
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| Institution: | Institute of Nuclear Agriculture Sciences, Zhejiang University, Key Laboratory of Nuclear Agricultural Sciences, Ministry of Agriculture of China;Institute of Nuclear Agriculture Sciences, Zhejiang University, Key Laboratory of Nuclear Agricultural Sciences, Ministry of Agriculture of China;Institute of Nuclear Agriculture Sciences, Zhejiang University, Key Laboratory of Nuclear Agricultural Sciences, Ministry of Agriculture of China;Institute of Nuclear Agriculture Sciences, Zhejiang University, Key Laboratory of Nuclear Agricultural Sciences, Ministry of Agriculture of China;Institute of Nuclear Agriculture Sciences, Zhejiang University, Key Laboratory of Nuclear Agricultural Sciences, Ministry of Agriculture of China;Institute of Nuclear Agriculture Sciences, Zhejiang University, Key Laboratory of Nuclear Agricultural Sciences, Ministry of Agriculture of China;Institute of Nuclear Agriculture Sciences, Zhejiang University, Key Laboratory of Nuclear Agricultural Sciences, Ministry of Agriculture of China;Institute of Nuclear Agriculture Sciences, Zhejiang University, Key Laboratory of Nuclear Agricultural Sciences, Ministry of Agriculture of China |
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| Abstract: | |
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| Keywords: | Arthrospira platensis Protein Preparation method Two-dimensional gel electrophoresis Helical chirality |
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