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中国精品科技期刊2020
地衣芽孢杆菌总蛋白双向电泳方法的建立和优化[J]. 食品工业科技, 2013, (15): 170-173. DOI: 10.13386/j.issn1002-0306.2013.15.061
引用本文: 地衣芽孢杆菌总蛋白双向电泳方法的建立和优化[J]. 食品工业科技, 2013, (15): 170-173. DOI: 10.13386/j.issn1002-0306.2013.15.061
Establishment and optimization of two-dimensional gel electrophoresis of total proteins from Bacillus lincheniformis[J]. Science and Technology of Food Industry, 2013, (15): 170-173. DOI: 10.13386/j.issn1002-0306.2013.15.061
Citation: Establishment and optimization of two-dimensional gel electrophoresis of total proteins from Bacillus lincheniformis[J]. Science and Technology of Food Industry, 2013, (15): 170-173. DOI: 10.13386/j.issn1002-0306.2013.15.061

地衣芽孢杆菌总蛋白双向电泳方法的建立和优化

Establishment and optimization of two-dimensional gel electrophoresis of total proteins from Bacillus lincheniformis

  • 摘要: 探索建立有效的地衣芽孢杆菌蛋白质组双向电泳体系,为进一步揭示地衣芽孢杆菌促进氧化葡萄糖酸杆菌产酸的作用机制奠定基础。以地衣芽孢杆菌为材料,比较蛋白质制备超声破壁时间、新型细胞裂解液、pH梯度和不同上样量对地衣芽孢杆菌蛋白双向电泳结果的影响。结果显示:采用15min超声破壁提取地衣芽孢杆菌总蛋白,选用新型蛋白质裂解,用长24cm、pH4~7的IPG胶条,在上样量为80μg进行等电聚焦,于60V15min、120V6h条件下进行SDS-PAGE垂直电泳,可以获得背景清晰、重复性好的双向电泳图谱。在探索出一种新型可行的新型细胞裂解液的同时,建立一套用于地衣芽孢杆菌蛋白质组分析的双向电泳方法。 

     

    Abstract: A two dimensional gel electrophoresis protocol proteomic study of Bacillus lincheniformis and offer further information how Bacillus cereus stimulate the growth of Gluconobacter oxydans to produce 2-keto-L- gulonic acid ( 2-KLG) after entering into stationary phase was established.Different parameters, including protein preparation by different ultrasonic broken time, new type lysis, pH gradient and different sample of Bacillus lincheniformis protein, were used to evaluate the effect of two - dimensional electrophoresis of Bacillus lincheniformis proteins. Results showed that the clear background and reproducibility of two dimensional gel electrophoresis were established on the condition of using 15min of ultrasonic broken extraction, selection of protein cleavage I, pH4 ~ 7 24cm IPG strips, 80μg loading volume, isoelectric focus at 60V 15min, 120V 6h.The aim of this study was not only to explore a novel cell lysate in two- dimensional gel electrophoresis, but also to establish a method of proteome analysis for two-dimensional electrophoresis of Bacillus licheniformis.

     

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