种子中Puroindoline蛋白的SDS-PAGE分析

SDS-PAGE Analysis of Puroindoline Protein in Seeds

Puroindoline蛋白是小麦中特殊的TritonX-114可溶性蛋白质,对小麦籽粒硬度有着决定性的影响。从二倍体、六倍体小麦材料及小麦近缘种粗山羊草成熟种子中提取Puroindoline蛋白,就对该蛋白的SDS-PAGE及染色条件进行了优化和讨论,建立了浓缩胶T(凝胶浓度)为4、C(交联度)为2.6、电泳电压为128V;分离胶T为13.5、C为2.6、电泳电压240V,分离胶电泳时间1.5h的结果稳定且重复性好的优化的SDS-PAGE条件。同时为降低电泳染色的实验成本,简化实验步骤,用蓝染法代替常用的银染法,并获得了良好的效果,为小麦中Puroindoline蛋白质的进一步研究奠定了实验基础。

Puroindoline protein is a special Triton X-114 soluble protein, and play critical roles in determining wheat grain hardness. Based on extracted puroindoline protein from diploid and hexaploid wheat and Aegilops tauschii, a optimized SDS-PAGE condition was built up with concentrated gel (T=4, C=2.6) voltage 128 V, separate gel (T=13.5, C=2.6) voltage 240 V to reach a more steady and repetitive electrophoresis results. At the same time,to reduce the test's cost and predigest the steps,blue-stained steps was used instead of silver-stained,and achieved an excellent results. The series tests therefore bulids foundarion for the farther study on puroindoline protein.