Isolation and in vitro translation of zein messenger ribonucleic acid

Abstract

Zein messenger RNA was isolated from membrane-bound polyribosomes of developing maize kernels by oligo(dT)-cellulose chromatography. Translation of the mRNA in vitro yielded protein similar to native zein in amino acid content, ethanol solubility, and mobility on sodium dodecyl sulfate- polyacrylamide gels. The zein mRNA sedimented as a homogeneous peak on sucrose gradients and contained a poly(A)-rich region based upon hybridization to [3H]poly(U). The mRNA had an apparent molecular weight of 540 000 on agarose-acrylamide gels. It synthesized both 21 800 and 19 000 molecular weight zein components in the wheat-germ cell-free protein synthesis system. The possibility of a polycistronic mRNA or two mRNAs of similar molecular weight is considered.