摘要
本文总结了我们在用淀粉凝胶电泳技术进行等位酶分析的实验过程中的一些经验和教训，主要结论是：１）尽管材料不同，对于不同的酶系统，用特定的缓冲液系统常能获得比较理想的效果，如ＤＩＡ，ＧＰＩ，ＨＥＸ，ＳＯＤ和ＴＰＩ用Ｓ６（Ｓｏｌｔｉｓ等，１９８３）；ＡＭＰ用Ｓ８；ＭＤＨ用Ｓ９和ＳＫＤ用Ｗ２（Ｗｅｎｄｅｌ和Ｗｅｅｄｅｎ，１９８９）等。２）用磷酸缓冲液配制ＡＭＰ的染色配方较其它配方效果好。３）当胶染时，使用琼脂替代琼脂糖，染色效果不变而成本更低，谱带扩散更慢。４）不同的提取液可能适用于不同的酶系统，如磷酸ＰＶＰ提取液（Ｓｏｌｔｉｓ等，１９８３）不适用于石荠苎的ＧＰＩ的提取。５）正确选择凝胶和电极缓冲液，Ｓ１虽然适用面广，但效果不一定好，对于位点多和等位基因丰富的材料尤其应警惕，因Ｓ１可能掩盖样品之间的差异。
This paper summarized our experience and lessons while we were doing starch gel electrophoresis experiment for allozyme analysis. High resolution was often achieved on some enzymes with certain buffer systems, for example, DIA,GPI,HEX,SOD and TPI with S6 (Soltis et al.,1983); AMP with S8;MDH with S9; and SKD with W2 (Wendel and Weeden,1989). Staining recipe of Soltis and Reiseberg (1986) was recommended for visualizing AMP. Agar staining was preferred to agarose staining for the reason of lower cost and slower diffusion of bands. For GPI,special attention should be paid to the choice of extraction buffer. In some cases, the bands of GPI were significantly reduced using the phosphate grinding buffer of Soltis et al. (1983) in some groups such as Mosla. The use of gel and electrode buffer No.1 (S1) should be avoided if the materials are isozymes and alleles rich.
出处
《植物学通报》
CSCD
1998年第5期68-72,共5页
Chinese Bulletin of Botany
基金
国家自然科学基金
中国科学院生物区系与分类特别支持费资助
