Plotnikov V K, Bakaldina N B
Laboratory of Molecular Biology, Krasnodar Lukyanenko Research Institute of Agriculture, Russia.
Plant Mol Biol. 1996 Jun;31(3):507-15. doi: 10.1007/BF00042224.
The lifetime of the zein mRNA in a developing corn (Zea mays L.) kernel under genome transcription blockade with actinomycin D (in vivo) and in a cell-free system (in vitro) was studied. After a 10 h blockade of gene transcription with actinomycin D, only 55% of 19 kDa zein mRNA and 40% of 22 kDa mRNA were detected in a developing kernel of normal corn. In that of the opaque-2 mutant 80% of 19 kDa zein mRNA remained. To examine the relative stability of poly(A)-containing mRNA, cell-free systems from rabbit reticulocyte lysate and wheat-germ extract were used. In both cases only 40% of 19 kDa zein mRNA and 60% of 22 kDa zein mRNA decayed during a 30 min incubation. Differential mRNA degradation of poly(A)-containing zein mRNA was observed on affinity chromatography; poly(A)-containing 19 kDa zein mRNA from normal corn partially decayed by elution from poly(U)-Sepharose whereas that from opaque-2 remained stable. These data suggest that differential mRNA stability is an important factor in the regulation of the zein gene expression in a developing corn kernel.
研究了在放线菌素D介导的基因组转录阻断条件下,发育中的玉米(Zea mays L.)籽粒中玉米醇溶蛋白mRNA在体内以及无细胞体系中(体外)的寿命。在用放线菌素D阻断基因转录10小时后,在正常玉米发育中的籽粒中仅检测到55%的19 kDa玉米醇溶蛋白mRNA和40%的22 kDa mRNA。在不透明-2突变体的籽粒中,80%的19 kDa玉米醇溶蛋白mRNA得以保留。为了检测含poly(A)的mRNA的相对稳定性,使用了来自兔网织红细胞裂解物和小麦胚芽提取物的无细胞体系。在这两种体系中,在30分钟的孵育过程中,仅40%的19 kDa玉米醇溶蛋白mRNA和60%的22 kDa玉米醇溶蛋白mRNA发生降解。在亲和层析中观察到含poly(A)的玉米醇溶蛋白mRNA的差异降解;来自正常玉米的含poly(A)的19 kDa玉米醇溶蛋白mRNA在从聚尿苷琼脂糖上洗脱时部分降解,而来自不透明-2突变体的该mRNA则保持稳定。这些数据表明,mRNA稳定性差异是发育中的玉米籽粒中玉米醇溶蛋白基因表达调控的一个重要因素。