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甘薯 SRD1 启动子在拟南芥、胡萝卜和马铃薯中赋予强烈的根、主根和块茎特异性表达。

A sweetpotato SRD1 promoter confers strong root-, taproot-, and tuber-specific expression in Arabidopsis, carrot, and potato.

机构信息

School of Life Sciences and Biotechnology, Korea University, Seoul, 136-701, Korea.

出版信息

Transgenic Res. 2012 Apr;21(2):265-78. doi: 10.1007/s11248-011-9528-4. Epub 2011 Jun 10.

Abstract

Harvestable, starch-storing organs of plants, such as fleshy taproots and tubers, are important agronomic products that are also suitable target organs for use in the molecular farming of recombinant proteins due to their strong sink strength. To exploit a promoter directing strong expression restricted to these storage organs, we isolated the promoter region (3.0 kb) of SRD1 from sweetpotato (Ipomoea batatas cv. 'White Star') and characterized its activity in transgenic Arabidopsis, carrot, and potato using the β-glucuronidase (GUS) gene (uidA) as a reporter gene. The SRD1 promoter conferred root-specific expression in transgenic Arabidopsis, with SRD1 promoter activity increasing in response to exogenous IAA. A time-course study of the effect of IAA (50 μM) revealed a maximum increase in SRD1 promoter activity at 24 h post-treatment initiation. A serial 5' deletion analysis of the SRD1 promoter identified regions related to IAA-inducible expression as well as regions containing positive and negative elements, respectively, controlling the expression level. In transgenic carrot, the SRD1 promoter mediated strong taproot-specific expression, as evidenced by GUS staining being strong in almost the entire taproot, including secondary phloem, secondary xylem and vascular cambium. The activity of the SRD1 promoter gradually increased with increasing diameter of the taproot in the transgenic carrot and was 10.71-fold higher than that of the CaMV35S promoter. The SRD1 promoter also directed strong tuber-specific expression in transgenic potato. Taken together, these results demonstrate that the SRD1 promoter directs strong expression restricted to the underground storage organs, such as fleshy taproots and tubers, as well as fibrous root tissues.

摘要

植物中可收获的、储存淀粉的器官,如肉质直根和块茎,是重要的农业产品,由于其强大的汇强度,它们也是用于重组蛋白的分子农业的合适目标器官。为了利用指导这些储存器官中强表达的启动子,我们从甘薯(Ipomoea batatas cv. 'White Star')中分离了 SRD1 的启动子区域(3.0 kb),并使用β-葡萄糖醛酸酶(GUS)基因(uidA)作为报告基因,在转基因拟南芥、胡萝卜和马铃薯中对其活性进行了表征。SRD1 启动子在转基因拟南芥中赋予根特异性表达,SRD1 启动子活性随外源 IAA 的增加而增加。IAA(50 μM)作用的时间进程研究表明,处理起始后 24 h 时 SRD1 启动子活性最大增加。SRD1 启动子的串联 5'缺失分析确定了与 IAA 诱导表达相关的区域,以及分别包含正和负调节元件的区域,控制表达水平。在转基因胡萝卜中,SRD1 启动子介导了强烈的肉质直根特异性表达,GUS 染色在几乎整个直根中均强烈,包括次生韧皮部、次生木质部和维管束形成层。在转基因胡萝卜中,SRD1 启动子的活性随着直根直径的增加而逐渐增加,其活性比 CaMV35S 启动子高 10.71 倍。SRD1 启动子还在转基因马铃薯中指导强烈的块茎特异性表达。总之,这些结果表明,SRD1 启动子指导强烈的表达仅限于地下储存器官,如肉质直根和块茎,以及纤维根组织。

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