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为减少小麦蛋白质分析的时间和提高分辨力,研究了毛细管长度、毛细管内径、缓冲液、温度、电压及提取条件等毛细管区带电泳(CZE)的参量。分析时间最短的条件是毛细管内径为20μm;尽量缩短毛细管为27cm(通向检测器为20cm);温度为45℃;电压为22kv;压力注入时间为4秒(0.25nl)。为了提高麦醇溶蛋白的提取和分析效果,用乙醇—水的3种溶剂系统试验,结果是30%乙醇—水这一系统最佳。从红皮硬粒冬小麦、红皮硬粒春小麦和软粒小麦种类的典型栽培种中提取麦醇溶谷蛋白,然后用CZE法分离。用CZE法对聚丙烯酰胺凝胶电泳在PH3.1下不能鉴别的3组栽培种进行鉴定,每组在10分钟以内就鉴定完毕。很多近缘(姊妹系或杂交种)栽培种也容易鉴定。遗传上无亲缘关系的栽培种,其CZE图谱差别很大。
To reduce the time and resolution of wheat protein analysis, capillary zone electrophoresis (CZE) parameters such as capillary length, capillary inner diameter, buffer, temperature, voltage and extraction conditions were studied. The shortest analysis time is the capillary inner diameter of 20μm; try to shorten the capillary 27cm (leading to the detector 20cm); temperature of 45 ℃; voltage of 22kv; pressure injection time of 4 seconds (0.25nl). In order to improve the extraction and analysis of gliadin, a system of 30% ethanol-water was the best system with three solvent systems of ethanol-water. Gliadin was extracted from typical cultivars of red hard durum winter wheat, red hulled spring wheat and soft wheat varieties and then separated by CZE method. CZE method polyacrylamide gel electrophoresis in PH3.1 can not be identified under three groups of cultivars were identified within 10 minutes each group was identified. Many closely related (sister or hybrid) cultivars are also easy to identify. Genetically unrelated cultivars vary greatly in their CZE patterns.