Ortiz de Bertorelli L, Ramírez A
Facultad de Agronomía, Universidad Central de Venezuela.
Arch Latinoam Nutr. 2000 Mar;50(1):74-80.
Polyacrylamide gel electrophoresis (PAGE) and PAGE-SDS were used to study seed albumins and globulins of Canavalia ensiformis. In PAGE the concentration of acrylamide used in the upper gel was 4.0% with a pH of 6.7 and in the lower gel 7.5% and 10% of acrylamide were used with a pH of 8.9. In PAGE-SDS the concentration of acrylamide was 4.4% with a pH of 6.8 in the upper gel and 7.5% and 12.6% with a pH of 8.8 in the lower gel. The material used were the Venezuelan genotypes Tovar, Yaracuy, Original and U-02. The albumins and globulins were extracted with a 0.5 M NaCl solution and then separated by dialysis against water and lyophilized. These protein fractions represented 84.85% of the total amount of protein in seeds. The albumins were separated in PAGE with 7.5% acrylamide into five fractions and globulins into six, with similar electrophoretic patterns between genotypes. In a similar manner, the patterns obtained in PAGE with 10% acrylamide were the same for all genotypes, showing five bands for albumins and three bands for globulins. With PAGE-SDS containing 7.5% of acrylamide, albumins were separated into as many as eight components, and globulins into as many as seven bands with mobilities between 0.2981 and 0.9932, with different patterns for each genotype. Also the patterns PAGE-SDS at 12.6% of acrylamide were different for the genotypes, separating proteins into a greater number of bands. The albumins showed as many as twenty-one bands with mobilities between 0.2603 and 0.7398, and globulins as many as sixteen bands with mobilities between 0.2454 and 0.7390. The PAGE patterns of the genotypes analyzed did not show differences between them. However, with PAGE-SDS different electrophoretic patterns were obtained which varied in the number and intensity of the bands, making it possible to distinguish the genotypes studied. The molecular weight of the albumins varied between 76,000 and 12,000 daltons and of the globulins between 80,000 and 12,000 daltons.
采用聚丙烯酰胺凝胶电泳(PAGE)和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(PAGE-SDS)对刀豆种子清蛋白和球蛋白进行研究。在PAGE中,上层凝胶中丙烯酰胺浓度为4.0%,pH值为6.7,下层凝胶中丙烯酰胺浓度分别为7.5%和10%,pH值为8.9。在PAGE-SDS中,上层凝胶中丙烯酰胺浓度为4.4%,pH值为6.8,下层凝胶中丙烯酰胺浓度分别为7.5%和12.6%,pH值为8.8。所用材料为委内瑞拉基因型托瓦尔、亚拉库伊、原始种和U-02。清蛋白和球蛋白用0.5M NaCl溶液提取,然后通过对水透析分离并冻干。这些蛋白质组分占种子总蛋白量的84.85%。在PAGE中,用7.5%丙烯酰胺可将清蛋白分离成五个组分,球蛋白分离成六个组分,不同基因型间电泳图谱相似。同样,在PAGE中用10%丙烯酰胺得到的图谱对所有基因型都相同,清蛋白显示五条带,球蛋白显示三条带。在含7.5%丙烯酰胺的PAGE-SDS中,清蛋白可分离成多达八个组分,球蛋白可分离成多达七条带,迁移率在0.2981至0.9932之间,不同基因型图谱不同。同样,在含12.6%丙烯酰胺的PAGE-SDS中,不同基因型图谱也不同,可将蛋白质分离成更多条带。清蛋白显示多达21条带,迁移率在0.2603至0.7398之间,球蛋白显示多达16条带,迁移率在0.2454至0.7390之间。所分析基因型的PAGE图谱未显示出差异。然而,PAGE-SDS得到了不同的电泳图谱,条带数量和强度各不相同,从而能够区分所研究的基因型。清蛋白分子量在76,000至12,000道尔顿之间,球蛋白分子量在80,000至12,000道尔顿之间。