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转基因植物中海牧豆球蛋白α'和β亚基基因的差异表达。

Differential expression of conglycinin α' and β subunit genes in transgenic plants.

机构信息

Plant Biology Program, Department of Biology, Washington University, 63130, St. Louis, MO, USA.

出版信息

Plant Mol Biol. 1988 Mar;11(2):109-23. doi: 10.1007/BF00015664.

Abstract

β-conglycinin or 7S protein, one of the two major storage proteins of soybean (Glycine max), is comprised of three subunits, α', α and β. Genomic clones encoding the α' and β subunit genes were ligated together in intermediate plasmids, either in direct orientation (← ←) or in divergent orientation (← →). In the latter instance the transcriptional promoters of the two genes were approximately 2.0 kb apart. The DNAs were introduced into leaf disks of Petunia hybrida on a disarmed Ti plasmid in Agrobacterium tumefaciens and transgenic plants were regenerated. Expression of the introduced genes, limited to specific stages in developing seeds, was analyzed by quantitating the accumulated mRNA and protein, and compared with expression in transgenic plants that contained either gene alone. Development time course studies showed that the α' subunit mRNA and protein were first detected at 10 days after pollination while accumulation of the β subunit protein ensued at day 14 regardless of the orientation of the two genes. The temporally regulated expression of these two genes in transgenic plants mimics their temporal regulation during soybean embryo development. In transgenic plants that contain the α' and β genes in divergent orientation, expression of the β gene was enhanced several-fold, while the expression of the α' gene was slightly decreased. These results suggest that (i) sequences of the α' gene enhance the level of expression of a nearby storage protein gene and that (ii) the signal(s) operating in the temporal regulation of seed storage protein genes is (are) common to soybean and petunia, and are different from those that govern the final amounts of mRNA and protein.

摘要

β-伴球蛋白或 7S 蛋白是大豆(Glycine max)两种主要贮藏蛋白之一,由三个亚基,即α'、α 和β组成。α'和β亚基基因的基因组克隆被连接到中间质粒上,要么是直接取向(← ←),要么是发散取向(← →)。在后一种情况下,两个基因的转录启动子大约相隔 2.0kb。将这些 DNA 导入到根癌农杆菌携带的失活 Ti 质粒的矮牵牛叶片圆盘中,并再生出转基因植物。通过定量分析积累的 mRNA 和蛋白质来分析引入基因的表达,仅限于发育种子的特定阶段,并与仅含有单个基因的转基因植物的表达进行比较。发育时间过程研究表明,α'亚基 mRNA 和蛋白质在授粉后 10 天首次检测到,而β亚基蛋白的积累则在第 14 天发生,无论两个基因的取向如何。这两个基因在转基因植物中的时间调节表达模拟了它们在大豆胚胎发育过程中的时间调节。在含有发散取向的α'和β基因的转基因植物中,β基因的表达增强了几倍,而α'基因的表达略有下降。这些结果表明:(i)α'基因的序列增强了附近贮藏蛋白基因的表达水平;(ii)调控种子贮藏蛋白基因时间表达的信号在大豆和矮牵牛中是相同的,与调控 mRNA 和蛋白质最终数量的信号不同。

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