适于双向电泳分析的水稻纹枯病菌菌体蛋白提取方法的比较
为建立水稻纹枯病菌菌体蛋白的双向电泳技术,获得分辨度高、重复性好的双向电泳图谱。采用了酚抽提法、TCA/丙酮沉淀法和丙酮沉淀法进行水稻纹枯病菌菌体蛋白质的提取,利用17 cm、pH 5～8的IPG胶条对样品进行双向电泳,用PDQuest 8.0软件对电泳图谱进行分析,确定电泳的最佳参数。结果显示：采用17 cm、pH 5～8的IPG胶条能得到较好的双向电泳图谱;同一样品的三种不同蛋白提取方法（酚抽提法、TCA/丙酮沉淀法和丙酮沉淀法）分别得到1278、885、827个蛋白质点。酚抽提法能有效去除样品中的盐分和小分子的干扰,得到背景清晰的蛋白图谱;TCA/丙酮沉淀法和丙酮沉淀法所得图谱有一定的杂...
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| Published in | 西南农业学报 Vol. 24; no. 6; pp. 2160 - 2163 |
|---|---|
| Main Author | |
| Format | Journal Article |
| Language | Chinese |
| Published |
广西大学,广西南宁,530005%广西农业科学院微生物研究所,广西南宁,530007%广西作物遗传改良生物技术重点开放实验室,广西南宁,530007%南阳师范学院生命科学与技术学院,河南南阳,473061
2011
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| Subjects | |
| Online Access | Get full text |
| ISSN | 1001-4829 |
| DOI | 10.3969/j.issn.1001-4829.2011.06.028 |
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| Abstract | 为建立水稻纹枯病菌菌体蛋白的双向电泳技术,获得分辨度高、重复性好的双向电泳图谱。采用了酚抽提法、TCA/丙酮沉淀法和丙酮沉淀法进行水稻纹枯病菌菌体蛋白质的提取,利用17 cm、pH 5～8的IPG胶条对样品进行双向电泳,用PDQuest 8.0软件对电泳图谱进行分析,确定电泳的最佳参数。结果显示：采用17 cm、pH 5～8的IPG胶条能得到较好的双向电泳图谱;同一样品的三种不同蛋白提取方法（酚抽提法、TCA/丙酮沉淀法和丙酮沉淀法）分别得到1278、885、827个蛋白质点。酚抽提法能有效去除样品中的盐分和小分子的干扰,得到背景清晰的蛋白图谱;TCA/丙酮沉淀法和丙酮沉淀法所得图谱有一定的杂质干扰,同时TCA/丙酮沉淀法和丙酮沉淀法有蛋白点缺失,尤其在碱性端存在明显缺失现象。研究结果建立了一套适于水稻纹枯病菌菌体蛋白的可靠提取方法和双向电泳体系,为进一步研究其蛋白质组学奠定了基础。 |
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| AbstractList | 为建立水稻纹枯病菌菌体蛋白的双向电泳技术,获得分辨度高、重复性好的双向电泳图谱。采用了酚抽提法、TCA/丙酮沉淀法和丙酮沉淀法进行水稻纹枯病菌菌体蛋白质的提取,利用17 cm、pH 5～8的IPG胶条对样品进行双向电泳,用PDQuest 8.0软件对电泳图谱进行分析,确定电泳的最佳参数。结果显示：采用17 cm、pH 5～8的IPG胶条能得到较好的双向电泳图谱;同一样品的三种不同蛋白提取方法（酚抽提法、TCA/丙酮沉淀法和丙酮沉淀法）分别得到1278、885、827个蛋白质点。酚抽提法能有效去除样品中的盐分和小分子的干扰,得到背景清晰的蛋白图谱;TCA/丙酮沉淀法和丙酮沉淀法所得图谱有一定的杂质干扰,同时TCA/丙酮沉淀法和丙酮沉淀法有蛋白点缺失,尤其在碱性端存在明显缺失现象。研究结果建立了一套适于水稻纹枯病菌菌体蛋白的可靠提取方法和双向电泳体系,为进一步研究其蛋白质组学奠定了基础。 S435.111.4; 为建立水稻纹枯病菌菌体蛋白的双向电泳技术,获得分辨度高、重复性好的双向电泳图谱.采用了酚抽提法、TCA/丙酮沉淀法和丙酮沉淀法进行水稻纹枯病菌菌体蛋白质的提取,利用17cm、pH 5 ～8的IPG胶条对样品进行双向电泳,用PDQuest 8.0软件对电泳图谱进行分析,确定电泳的最佳参数.结果显示:采用17cm、pH 5 ～8的IPG胶条能得到较好的双向电泳图谱；同一样品的三种不同蛋白提取方法(酚抽提法、TCA/丙酮沉淀法和丙酮沉淀法)分别得到1278、885、827个蛋白质点.酚抽提法能有效去除样品中的盐分和小分子的干扰,得到背景清晰的蛋白图谱；TCA/丙酮沉淀法和丙酮沉淀法所得图谱有一定的杂质干扰,同时TCA/丙酮沉淀法和丙酮沉淀法有蛋白点缺失,尤其在碱性端存在明显缺失现泉.研究结果建立了一套适于水稻纹枯病菌菌体蛋白的可靠提取方法和双向电泳体系,为进一步研究其蛋白质组学奠定了基础. |
| Author | 余功明 胡春锦 魏源文 史国英 岑贞陆 覃丽萍 黎起秦 黄思良 |
| AuthorAffiliation | 广西大学,广西南宁530005 广西农业科学院微生物研究所,广西南宁530007 广西作物遗传改良生物技术熏点开放实验室,广西南宁530007 南阳师范学院生命科学与技术学院,河南南阳473061 |
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| Author_FL | LI Qi-qin SHI Guo-ying YU Gong-ming HU Chun-jin QIN Li-ping CEN Zhen-lu HUANG Si-liang WEI Yuan-wen |
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| ClassificationCodes | S435.111.4 |
| ContentType | Journal Article |
| Copyright | Copyright © Wanfang Data Co. Ltd. All Rights Reserved. |
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| DOI | 10.3969/j.issn.1001-4829.2011.06.028 |
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| DocumentTitleAlternate | Efficient Protein Extraction Method from Rhizoctonia solani for Proteomic Analysis Using Two-dimensional Electrophoresis Analysis |
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| Keywords | 双向电泳 水稻纹枯病菌 提取方法 总蛋白 |
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| Notes | YU Gong-ming,HU Chun-jin,WEI Yuan-wen,SHI Guo-ying,CEN Zhen-lu,QIN Li-ping,LI Qi-qin,HUANG Si-liang（1.Guangxi University,Guangxi Nanning 53005,China;2.Microbiology Institute,Guangxi Academy of Agricultural Sciences,Guangxi Nanning 530007,China;3.Guangxi Crop Genetic Improvement and Biotechnology Lab,Guangxi Nanning 530007,China;4.College of Life Sciences and Technology,Nanyang Normal University,Henan Nanyang 473061,China） 51-1213/S To obtain clear two-dimensional electrophoresis（2-DE） image with highest resolution of Rhizoctonia solani,three protein extraction protocols were compared.After analyzing the 2-DE protein images by the software of PDQuest8.0,1278,885and827 protein spots were obtained separately by the methods of phosphate solubilization,TCA/acetone precipitation and acetone precipitation,the IPG strip is pH 5-8,17 cm.2-DE images with clearer background were observed and more protein spots were obtained by phosphate solubilization compared to other protocols,which was due to effective removal of salt |
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| PublicationYear | 2011 |
| Publisher | 广西大学,广西南宁,530005%广西农业科学院微生物研究所,广西南宁,530007%广西作物遗传改良生物技术重点开放实验室,广西南宁,530007%南阳师范学院生命科学与技术学院,河南南阳,473061 |
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| Snippet | 为建立水稻纹枯病菌菌体蛋白的双向电泳技术,获得分辨度高、重复性好的双向电泳图谱。采用了酚抽提法、TCA/丙酮沉淀法和丙酮沉淀法进行水稻纹枯病菌菌体蛋白质的提取,利用17... S435.111.4; 为建立水稻纹枯病菌菌体蛋白的双向电泳技术,获得分辨度高、重复性好的双向电泳图谱.采用了酚抽提法、TCA/丙酮沉淀法和丙酮沉淀法进行水稻纹枯病菌菌体蛋白质的... |
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| SubjectTerms | 双向电泳 总蛋白 提取方法 水稻纹枯病菌 |
| Title | 适于双向电泳分析的水稻纹枯病菌菌体蛋白提取方法的比较 |
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