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首页> 外文期刊>Cytology and genetics >Comparison of gfp Gene Expression Levels after Agrobacterium- Mediated Transient Transformation of Nicotiana rustica L. by Constructs with Different Promoter Sequences
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Comparison of gfp Gene Expression Levels after Agrobacterium- Mediated Transient Transformation of Nicotiana rustica L. by Constructs with Different Promoter Sequences

机译：用不同启动子序列的构建体介导尼古拉毒素L.瞬态转化后GFP基因表达水平的比较

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摘要

Promoters are key elements regulating gene expression levels, therefore their selection is an important step in genetic engineering research. The reporter gene gfp, which encodes green fluorescent protein (GFP), was transiently expressed in leaf tissues of Aztec tobacco Nicotiana rustica L. Compared to other species of the Nicotiana genus, Aztec tobacco has a large potential for expression of heterologous proteins, a large vegetative biomass, can be easily infiltrated, and is unpretentious in cultivation. Six genetic constructs were used with different promoter sequences: the 35S promoter of Cauliflower Mosaic Virus (35S CaMV), the double-enhanced 35S promoter (D35S CaMV), promoters of the RbcS1B and RbcS2B genes encoding the small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) isolated from Arabidopsis thaliana (L.) Heynh., and promoters of the LHB1B1 and LHB1B2 genes from A. thaliana encoding chlorophyll a/b binding proteins. The gfp gene expression was detected visually, spectrofluorimetrically, and by protein content (Bradford assay) on the seventh day after infiltration. The highest level of expression was observed using the double-enhanced 35S promoter (D35S CaMV) and the lowest using the LHB1B1 gene promoter.
机译：启动子是调节基因表达水平的关键元素，因此它们的选择是基因工程研究的重要一步。编码绿色荧光蛋白（GFP）的报告基因GFP，在阿兹特克烟草烟草菌霉菌L的叶组织中瞬时表达。与其他种类的烟草属，Aztec烟草具有大量的异源蛋白表达，大量植物生物量，可以很容易地渗透，培养方面是不稳定的。六种遗传构建体与不同的启动子序列一起使用：花椰菜马赛克病毒（35s Camv）的35s启动子，双增强型35s启动子（D35S Camv），RBCS1B的启动子和编码核苷酸的小亚基的RBCS2B基因-1,5 - 从拟南芥（L.）HeynA（LHB1B1和LHB1B2基因的亚麻磷酸羧糖酶/氧气（Rubisco）和来自A.植物编码叶绿素A / B结合蛋白的启动子。在渗透后第七天，在视觉上，光谱氟化学性和通过蛋白质含量（Bradford测定）检测到GFP基因表达。使用双增强型35s启动子（D35S CAMV）和使用LHB1B1基因启动子最低观察到最高水平的表达。

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