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玉米中编码玉米醇溶蛋白多肽的基因和信使核糖核酸。

Genes and mRNAs coding for zein polypeptides in Zea mays.

作者信息

Viotti A, Sala E, Marotta R, Alberi P, Balducci C, Soave C

出版信息

Eur J Biochem. 1979 Dec;102(1):211-22. doi: 10.1111/j.1432-1033.1979.tb06282.x.

Abstract

Zein messenger RNAs from maize endosperm were purified by successive oligo(dT)-cellulose chromatography and sucrose gradient centrifugation. Polyacrylamide gel electrophoresis under denaturing conditions revealed the presence of two size classes of zein messenger RNAs of Mr 3.5 x 10(5) and 4.10 x 10(5). The mRNA was shown to synthesize the major zein polypeptides, to have a base composition characteristic of a poly(A)-containing RNA and to be transcribed by reverse transcriptase into complementary DNA. The r0t1/2 of the hybridization curve of cDNA hybridized to an excess of mRNA was shown to be 7 x 10(-2) M . 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 2 n cells suggest a ten-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.

摘要

来自玉米胚乳的玉米醇溶蛋白信使核糖核酸(mRNA)通过连续的寡聚(dT)-纤维素柱层析和蔗糖梯度离心进行纯化。变性条件下的聚丙烯酰胺凝胶电泳显示存在两种大小类别的玉米醇溶蛋白mRNA,其分子量分别为3.5×10⁵和4.10×10⁵。该mRNA被证明能合成主要的玉米醇溶蛋白多肽,具有含聚腺苷酸(poly(A))RNA的碱基组成特征,并能被逆转录酶转录成互补DNA(cDNA)。与过量mRNA杂交的cDNA杂交曲线的半反应时间(r0t1/2)为7×10⁻²M·s,表明玉米醇溶蛋白mRNA制剂中存在约15个非交叉杂交序列。cDNA与来自二倍体细胞的过量玉米DNA退火的动力学表明每个mRNA序列重复了十次。这一结果在饱和实验中得到证实,即在cDNA过量于DNA的情况下,单倍体玉米基因组中玉米醇溶蛋白基因的数量估计约为120个拷贝。对正常和突变胚乳(三倍体细胞)的DNA进行的类似实验表明,在专门用于玉米醇溶蛋白合成的组织中,玉米醇溶蛋白基因不存在大量扩增或缺失。

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