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Position: Home > Articles > Optimization of Two-dimensional Polyaneylamide Gel Electrophoresis for Proteome of Kandelia candel Roots Fujian Journal of Agricultural Sciences 2012,27 (8) 863-868

秋茄根系蛋白质组的双向电泳技术优化

作  者:
陈伟;黄榕辉;林小敏;杨镇鸿;翁兆霞;谭芳林
单  位:
福建农林大学生命科学学院;福建省林业科学研究院
关键词:
秋茄;根系;双向电泳;蛋白质组
摘  要:
建立适用于秋茄根系蛋白质组分析的双向电泳技术,并对秋茄根系蛋白质的制备、溶解、上样量、IEF及SDS-PAGE电泳等关键步骤进行优化。结果表明,应用酚抽法来提取秋茄根系蛋白,更适用于双向电泳分析;秋茄根系蛋白主要分布在pH4~7范围;裂解液中含有硫脲(2mmol.L-1)能较充分地溶解蛋白,DTT浓度为40mmol.L-1、上样量1.5mg时得到的双向电泳图谱分辨率较好,且蛋白斑点分布均匀、清晰,拖尾现象明显减少。采用与质谱兼容的考马斯亮兰进行染色,得到1 089个蛋白点。
译  名:
Optimization of Two-dimensional Polyaneylamide Gel Electrophoresis for Proteome of Kandelia candel Roots
作  者:
CHEN Wei1,HUANG Rong-hui1,LIN Xiao-min1,YANG Zhen-hong1,WENG Zhao-xia1,TAN Fang-lin2(1.School of Life Sciences,Fujian Agriculture and Forestry University,Fuzhou,Fujian 350002,China; 2.Fujian Academy of Forestry,Fuzhou,Fujian 350002,China)
关键词:
Kandelia candel;roots;Two-dimensional polyareylamide gel electrophoresis;Proteome
摘  要:
A two-dimensional polyarelamide gel electrophoresis(2D-PAGE) system was established for proteomic analysis of Kandelia candel roots.Protein extraction,dissolution,loading quantity,IEF and SDS-PAGE parameters in the two-dimensional electrophoresis(2-DE) were optimized.The results showed that phenol method was suitable for the root protein extraction,the protein isoelectric points mainly distributed between pH 4 and 7.The lysis buffer with 2 mmol·L-1 thiourea improved dissolution of protein effectively and the resolution of spectrum for the dissolved protein was increased with 40 mmol·L-1 DTT and 1.5 mg loading amount.The 1089 protein spots were stained with Colloidal coomassie blue and detected by mass spectrometry.

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