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编码一种假定的7 kDa脂质转移蛋白的大麦糊粉层特异性基因的启动子,在转基因水稻中赋予糊粉层细胞特异性表达。

The promoter of the barley aleurone-specific gene encoding a putative 7 kDa lipid transfer protein confers aleurone cell-specific expression in transgenic rice.

作者信息

Kalla R, Shimamoto K, Potter R, Nielsen P S, Linnestad C, Olsen O A

机构信息

Department of Biotechnological Sciences, Agricultural University of Norway.

出版信息

Plant J. 1994 Dec;6(6):849-60. doi: 10.1046/j.1365-313x.1994.6060849.x.

Abstract

This paper describes the aleurone-specific gene Ltp2 from barley, which encodes a putative 7 kDa non-specific lipid transfer protein. As shown by Northern and in situ hybridization analyses, the Ltp2 transcript is present in barley aleurone cells shortly after the initiation of aleurone cell differentiation. The expression of Ltp2 increases until grain mid-maturity, but the mRNA is absent from mature grains. The Ltp2 transcript is undetectable in the embryo and vegetative tissues, confirming the aleurone specificity of the Ltp2 gene. The ability of the isolated 801 bp Ltp2 promoter to direct aleurone-specific expression in immature barley grains is demonstrated by particle bombardment experiments. In these experiments, the activity of the Ltp2 promoter is 5% of the activity of the strong constitutive Actin1 promoter from rice, as quantified by GUS activity measurements. In stably transformed rice plants containing the Ltp2 promoter-Gus construct, the specificity of the Ltp2 promoter is confirmed in vivo by the presence of GUS activity exclusively in the aleurone layer. This study demonstrates the conserved nature of the regulatory signals involved in aleurone-specific gene transcription in cereal grains.

摘要

本文描述了大麦中糊粉层特异性基因Ltp2,该基因编码一种推测的7 kDa非特异性脂质转运蛋白。Northern杂交和原位杂交分析表明,Ltp2转录本在糊粉层细胞分化开始后不久就存在于大麦糊粉层细胞中。Ltp2的表达一直增加到籽粒成熟中期,但在成熟籽粒中不存在该mRNA。在胚和营养组织中未检测到Ltp2转录本,证实了Ltp2基因的糊粉层特异性。通过粒子轰击实验证明了分离出的801 bp Ltp2启动子在未成熟大麦籽粒中指导糊粉层特异性表达的能力。在这些实验中,通过GUS活性测量定量,Ltp2启动子的活性是来自水稻的强组成型肌动蛋白1启动子活性的5%。在含有Ltp2启动子-Gus构建体的稳定转化水稻植株中,通过仅在糊粉层中存在GUS活性在体内证实了Ltp2启动子的特异性。这项研究证明了谷物中糊粉层特异性基因转录所涉及的调控信号的保守性质。

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