Electrophoretic Fractionation and Translation in vitro of Poly(rA)‐Containing RNA from Maize Endosperm

Abstract

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 trnaslation 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-M_r protein and one coding for the 19000-M_r protein.