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甘薯RbcS基因(IbRbcS1)启动子在转基因拟南芥中赋予GUS报告基因高水平且绿色组织特异性表达。

The sweet potato RbcS gene (IbRbcS1) promoter confers high-level and green tissue-specific expression of the GUS reporter gene in transgenic Arabidopsis.

作者信息

Tanabe Noriaki, Tamoi Masahiro, Shigeoka Shigeru

机构信息

Department of Advanced Bioscience, Faculty of Agriculture, Kinki University, Nakamachi, Nara 631-8505, Japan; Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency, Kawaguchi 332-0012, Japan.

Department of Advanced Bioscience, Faculty of Agriculture, Kinki University, Nakamachi, Nara 631-8505, Japan; Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency, Kawaguchi 332-0012, Japan.

出版信息

Gene. 2015 Aug 10;567(2):244-50. doi: 10.1016/j.gene.2015.05.006. Epub 2015 May 7.

Abstract

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 acid 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 β-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.

摘要

甘薯因其高产和生物量生产而成为一种重要的作物。我们在此研究了甘薯(Ipomoea batatas)中1,5-二磷酸核酮糖羧化酶/加氧酶小亚基(RbcS)启动子活性的潜力,以便开发拟南芥中外源DNA的高表达系统。我们从甘薯中分离出两个编码RbcS的不同cDNA(IbRbcS1和IbRbcS2)。它们预测的氨基酸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的表达特异性于绿色组织,而IbRbcS2的表达则特异性于根和块茎等非光合组织。这些结果表明,与IbRbcS2相比,IbRbcS1在甘薯的绿色组织中占主导地位表达。因此,将IbRbcS1启动子与作为报告基因的β-葡萄糖醛酸酶(GUS)一起转化到拟南芥中。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GUS报告基因表达。此外,qPCR表明,IbRbcS1 pro:GUS中GUS报告基因的表达水平与CaMV 35S pro:GUS植物中的表达水平相同。这些结果表明,IbRbcS1启动子是一种潜在的强大的外源基因表达系统,用于植物的遗传转化。

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