Department of Biological Sciences, Purdue University, West Lafayette, Indiana 47907.
Plant Physiol. 1988 Dec;88(4):1002-7. doi: 10.1104/pp.88.4.1002.
Transcriptional fusions composed of a 19 kilodalton zein cDNA, the 5' flanking region from a beta-phaseolin gene, and 3' flanking regions from either the phaseolin or a 15-kilodalton zein gene were introduced into Petunia by Agrobacterium-mediated transformation. The expression of both zein mRNA and protein in these transgenic plants was seed-specific and developmentally regulated. Both monocot (zein) and dicot (phaseolin) polyadenylation consensus sequences were recognized in Petunia. Analysis by immunoblotting showed that the M(r) of the zein protein corresponded to that of the mature protein, suggesting that recognition and cleavage of the signal sequence had occurred. While zein mRNA accumulated to approximately 1% of the total poly(A)(+) RNA in seeds of the transformed plants, zein protein was present at a much lower concentration than expected, at most being 0.005% of the total seed protein. These results suggest that the 19 kilodalton zein gene, in addition to lacking specific sequences required for efficient transcription in dicots, might also lack sequences required for the efficient synthesis, targeting, transport, or stabilization of the protein.
转录融合物由一个 19 千道尔顿的玉米醇溶蛋白 cDNA、一个 β-伴球蛋白基因的 5'侧翼区以及一个来自伴球蛋白或 15 千道尔顿玉米醇溶蛋白基因的 3'侧翼区组成,通过根癌农杆菌介导转化被引入矮牵牛。这些转基因植物中玉米醇溶蛋白 mRNA 和蛋白的表达均具有种子特异性和发育调控性。在矮牵牛中,既识别单(玉米醇溶蛋白)又识别双(伴球蛋白)聚腺苷酸化的保守序列。免疫印迹分析表明,玉米醇溶蛋白的 M(r)与成熟蛋白相对应,这表明信号序列的识别和切割已经发生。虽然玉米醇溶蛋白 mRNA 在转化植物种子中的总多聚(A)+ RNA 中积累到约 1%,但玉米醇溶蛋白蛋白的含量却远低于预期,最多为种子总蛋白的 0.005%。这些结果表明,除了缺乏在双子叶植物中有效转录所需的特定序列外,19 千道尔顿的玉米醇溶蛋白基因可能还缺乏有效合成、靶向、运输或稳定蛋白所需的序列。