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| 双向电泳结合质谱初步分析蛇岛蝮蛇毒蛋白质组 | |
| 引用本文： | 马驰,刘淑清,宗军卫,袁章利,孙明忠,吴玉群,李建立,孙立新,郑庆印.双向电泳结合质谱初步分析蛇岛蝮蛇毒蛋白质组[J].动物学杂志,2009,44(4):70-77. |
| 作者姓名： | 马驰 刘淑清 宗军卫 袁章利 孙明忠 吴玉群 李建立 孙立新 郑庆印 |
| 作者单位： | 1. 大连医科大学临床医学系,大连,116044 2. 大连医科大学生化与分子生物学教研室,大连,116044 3. 大连医科大学基础医学院生物技术系,大连,116044 4. 辽宁蛇岛老铁山自然保护区管理处,大连,116041 5. 凯斯西保留地大学医学院,克利夫兰,44106,美国 |
| 基金项目： | 大连医科大学大学生科研活动资助项目;;辽宁省自然科学基金项目(No.2008S077);;大连科技厅资助项目(No.2008J22JH014);;辽宁省百千万人才工程资助项目(No.2008921069) |
| 摘 要： | 采用荧光染料(Cy5)标记中国辽宁蛇岛蝮(Gloydius shedaoensis shedaoensis,GSS)蛇毒(snake venom,SV,GSS-SV)蛋白质,获得了该蛇毒的双向十二烷基硫酸钠聚丙烯酰胺凝胶电泳(2D SDS-PAGE)图谱,经DeCyder软件分析,分辨出1000多个蛋白点,分子量范围在10～150ku间,等电点在4～7的蛋白质点占78.8%。凝胶后染色(post-staining)采用蛋白荧光染料Deep Purple,选取的5个蛋白点经胶内酶解,产生的肽段经高效液相色谱-电喷雾串联质谱(high performance liquid chromatography/HPLC-electro-spray ionization tandem mass spectrometry/ESI-MS/MS,HPLC-ESI-MS/MS)进行序列测定,质谱数据经Sequest Bioworks软件分析,为蛇毒L-氨基酸氧化酶、金属蛋白酶、类凝血酶、纤溶酶原激活物和磷脂酶A2的同源蛋白。本研究采用的荧光标记2DSDS-PAGE结合HPLC-ESI-MS/MS的技术适于高通量研究蛇毒蛋白组成。 |
| 关 键 词： | 蛇岛蝮 蛇毒 双向电泳 高效液相色谱 电喷雾质谱 |
| 收稿时间： | 2009/1/16 0:00:00 |
| 修稿时间： | 2009/4/23 0:00:00 |
Proteomic Analysis of Gloydius shedaoensis shedaoensis Venom by Two-dimensional Electrophoresis Combined to High Performance Liquid Chromatography-electro-spray Ionization Tandem Mass Spectrometry:a Preliminary Study |
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| MA Chi,LIU Shu-Qing,ZONG Jun-Wei,YUAN Zhang-Li,SUN Ming-Zhong,WU Yu-Qun,LI Jian-Li,SUN Li-Xin and ZHENG Qing-Yin.Proteomic Analysis of Gloydius shedaoensis shedaoensis Venom by Two-dimensional Electrophoresis Combined to High Performance Liquid Chromatography-electro-spray Ionization Tandem Mass Spectrometry:a Preliminary Study[J].Chinese Journal of Zoology,2009,44(4):70-77. | |
| Authors: | MA Chi LIU Shu-Qing ZONG Jun-Wei YUAN Zhang-Li SUN Ming-Zhong WU Yu-Qun LI Jian-Li SUN Li-Xin and ZHENG Qing-Yin |
| Institution: | Department of Clinic Medicine,Dalian Medical University,Dalian 116044;Department of Biochemistry & Molecular Biology,Dalian Medical University,Dalian 116044;Department of Biochemistry & Molecular Biology,Dalian Medical University,Dalian 116044;Department of Biochemistry & Molecular Biology,Dalian Medical University,Dalian 116044;Department of Biotechnology,Dalian Medical University,Dalian 116044;Administrative Division of the National Snake Island and Laotieshan Mountain Natural Conservation,Dalian 116041,China;Administrative Division of the National Snake Island and Laotieshan Mountain Natural Conservation,Dalian 116041,China;Administrative Division of the National Snake Island and Laotieshan Mountain Natural Conservation,Dalian 116041,China;Case Western Reserve University Medical School,Cleveland,Ohio 44106,USA |
| Abstract: | The protein components contained in the snake venom of Gloydius shedaoensis shedaoensis(GSS-SV) were labeled with the fluorescent dye of Cy5.Then,the two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis(2D SDS-PAGE) profile of GSS-SV proteome was obtained.Gel image analysis by DeCyder software resolved over 1 000 protein spots with the apparent molecular weights ranging between 10 and 150 ku on the gel.The protein spots with the pI ranging between 4 and 7 covered about 78.8% of the over... |
| Keywords: | Gloydius shedaoensis shedaoensis Snake venom Two-dimensional electrophoresis High performance liquid chromatography Electro-spray ionization tandem mass spectrometry |
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