首页> 外文期刊>Gene: An International Journal Focusing on Gene Cloning and Gene Structure and Function >The sweet potato RbcS gene (IbRbcS1) promoter confers high-level and green tissue-specific expression of the GUS reporter gene in transgenic Arabidopsis
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The sweet potato RbcS gene (IbRbcS1) promoter confers high-level and green tissue-specific expression of the GUS reporter gene in transgenic Arabidopsis

机译:甘薯RbcS基因(IbRbcS1)启动子赋予转基因拟南芥中GUS报告基因高水平和绿色组织特异性表达

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Sweet potato is an important crop because of its high yield and biomass production. We herein investigated the potential of the promoter activity of a small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase (RbcS) from sweet potato (Ipomoea batatas) in order to develop the high expression system of exogenous DNA in Arabidopsis. We isolated two different cDNAs (IbRbcS1 and IbRbcS2) encoding RbcS from sweet potato. Their predicted amino add sequences were well conserved with the mature RbcS protein of other plants. The tissue-specific expression patterns of these two genes revealed that expression of IbRbcS1 was specific to green tissue, whereas that of IbRbcS2 was non-photosynthetic tissues such as roots and tubers. These results suggested that IbRbcS1 was predominantly expressed in the green tissue-specific of sweet potato over IbRbcS2. Therefore, the IbRbcS1 promoter was transformed into Arabidopsis along with beta-glucuronidase (GUS) as a reporter gene. GUS staining and semi-quantitative RT-PCR showed that the IbRbcS1 promoter conferred the expression of the GUS reporter gene in green tissue-specific and light-inducible manners. Furthermore, qPCR showed that the expression levels of GUS reporter gene in IbRbcS1 pro:GUS were same as those in CaMV 35S pro:GUS plants. These results suggest that the IbRbcS1 promoter is a potentially strong foreign gene expression system for genetic transformation in plants. (C) 2015 Elsevier B.V. All rights reserved.
机译:甘薯因其高产量和生物量生产而成为重要的农作物。为了开发拟南芥中外源DNA的高表达系统,我们在本文中研究了甘薯(番薯(Ipomoea batatas))核糖-1,5-双磷酸羧化酶/加氧酶(RbcS)的小亚基的启动子活性的潜力。我们从甘薯中分离了两个不同的cDNA(IbRbcS1和IbRbcS2)编码RbcS。他们预测的氨基酸添加序列与其他植物的成熟RbcS蛋白高度保守。这两个基因的组织特异性表达模式表明,IbRbcS1的表达对绿色组织具有特异性,而IbRbcS2的表达则是非光合组织,例如根和块茎。这些结果表明,与IbRbcS2相比,IbRbcS1主要在甘薯的绿色组织特异性中表达。因此,IbRbcS1启动子与β-葡萄糖醛酸苷酶(GUS)一起作为报道基因被转化为拟南芥。 GUS染色和半定量RT-PCR表明,IbRbcS1启动子以绿色组织特异性和光诱导方式赋予了GUS报告基因表达。此外,qPCR显示IbRbcS1 pro:GUS中GUS报告基因的表达水平与CaMV 35S pro:GUS植物中的相同。这些结果表明,IbRbcS1启动子是植物遗传转化中潜在的强大外源基因表达系统。 (C)2015 Elsevier B.V.保留所有权利。

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