红肉蜜柚汁胞蛋白质双向电泳体系优化研究

Optimization of Two-Dimensional Gel Electrophoresis System for Analyzing ‘Hongroumiyou' Juice Sacs Proteins

为建立适合红肉蜜柚发育过程中汁胞蛋白质组学的研究体系,以着色初始时期的红肉蜜柚汁胞为试材,摸索最适合的蛋白质双向电泳体系参数。结果表明,酚提取法比TCA-丙酮法更适合于红肉蜜柚汁胞总蛋白质的提取,提取得率高,双向电泳图谱清晰。采取18 cm、线性pH 3～10和18 cm、线性pH 4～7相结合的分析策略,可获得更为完整的蛋白质组信息,其第一向等电聚焦最佳参数(总伏小时数)分别为30 000 Vhs和43 000 Vhs。

In order to establish proteomics research system for analyzing ＇Hongroumiyou＇ juice sacs proteins, using ＇Hongroumiyou＇ juice sacs in initial color change stage as test material, exploring the most suitable 2-DE system parameters. The results indicated that compared with TCA-acetone extraction method, phenol extraction method was more appropriate for ＇Hongroumiyou＇ juice sacs total protein yield. Using 18 cm, pH 4 - 7 linner immobiline combined with 18 cm, pH 3 -10 linner immobiline could had clear and high resolution 2-DE patterns. Both of the best isoelectric focused parameter was 43 000 vhs and 30 000 vhs, respectively.