Genes and mRNAs Coding for Zein Polypeptides in Zea mays

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Abstract
Zein mRNA from maize endosperm were purified by successive oligo(dT)-cellulose chromatography and sucrose gradient centrifugation. Polyacrylamide gel electrophoresis under denaturating conditions revealed the presence of 2 size classes of zein mRNA of MW 3.5 .times. 105 and 4.10 .times. 105. The mRNA synthesized the major zein polypeptides, had a base composition characteristic of a poly(A)-containing RNA and transcribed by reverse transcriptase into complementary [c] DNA. The rot1/2 [half-renaturation time] of the hybridization curve of cDNA hybridized to an excess of mRNA was 7 .times. 10-2 M .cntdot. s indicating that about 15 non-cross-hybridizing sequences are present in the zein mRNA preparations. The kinetics of cDNA annealing with an excess of maize DNA from 2n cells suggest a 10 times reiteration of each mRNA sequence. This result is confirmed from saturation experiments, where in cDNA excess to DNA, the number of zein genes per haploid maize genome was estimated as about 120 copies. Similar experiments carried out on DNA from normal and mutant endosperms (3n cells) indicate the absence of large amplifications or deletions of zein genes in the tissue devoted to zein synthesis.