Wienand U, Feix G
Eur J Biochem. 1978 Dec;92(2):605-11. doi: 10.1111/j.1432-1033.1978.tb12783.x.
In the search for individual mRNAs coding for particular zein proteins, polysomal RNA was isolated from the endosperm of 22-days post-pollination maize kernels. This RNA was enriched for poly(rA)-containing RNAs and then submitted to preparative polyacrylamide gel electrophoresis under denaturing conditions. After electrophoresis, RNA fractions were eluted from the polyacrylamide gel and analyzed for zein mRNA activity by translation in vitro in the wheat germ system. The wheat germ system had previously been optimized for accurate translation of zein mRNAs. By a gel-electrophoretic analysis of immunoprecipitated products from the translation reactions in vitro, it could conclusively be shown that the endosperm of developing maize kernels contains two separable mRNAs for zein, one coding for the 22000-Mr protein and one coding for the 19000-Mr protein.
在寻找编码特定玉米醇溶蛋白的单个信使核糖核酸(mRNA)的过程中,从授粉后22天的玉米籽粒胚乳中分离出多聚核糖体RNA。这种RNA富含含聚(A)的RNA,然后在变性条件下进行制备性聚丙烯酰胺凝胶电泳。电泳后,从聚丙烯酰胺凝胶中洗脱RNA组分,并通过在小麦胚芽系统中进行体外翻译来分析玉米醇溶蛋白mRNA活性。小麦胚芽系统先前已针对玉米醇溶蛋白mRNA的准确翻译进行了优化。通过对体外翻译反应中免疫沉淀产物的凝胶电泳分析,可以确凿地表明发育中的玉米籽粒胚乳含有两种可分离的玉米醇溶蛋白mRNA,一种编码22000道尔顿的蛋白质,另一种编码19000道尔顿的蛋白质。