Construction of efficient, tuber-Specific, and cold-inducible promoters in potato
Construction of efficient, tuber-Specific, and cold-inducible promoters in potato
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马铃薯高效、块茎特异性、冷诱导启动子的构建
DOI：
10.1016/j.plantsci.2015.02.014
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发表时间：
2015-06-01
期刊：
影响因子：
5.2
通讯作者：
Liu, Jun
中科院分区：
文献类型：
--
作者：
Li, Meng;Xie, Conghua;Liu, Jun
启动子活性是基因精确表达的关键。以前，人们构建了一个块茎特异的冷诱导启动子PCL，该启动子含有一个C-重复/脱水反应元件(CRT/DRE)盒和一个块茎特异片段，以调控马铃薯的冷诱导甜化(CIS)。然而，由于PCL活性较低，其应用受到了限制。为了提高其对低温的诱导能力，我们修改了CRT/DRE和侧翼序列。特别是，通过对CRT/DRE核心元件(CCGAC)旁的侧翼序列进行定点突变，显著提高了启动子的活性。我们还在PCL中插入了一个改良的CRT/DRE盒；尽管这种活性增强了，但它并不比侧翼序列的突变更有效。事实上，通过用两个突变的CRT/DRE盒的串联重复来取代PCL中的CRT/DRE盒，可以实现高达20倍的PCL活性增强。这一改进是由于CRT/DRE盒(S)与StCBF1转录因子之间的亲和力增强。综上所述，这些数据表明改变CRT/DRE的结构可以促进CBF相关转录复合体的形成，从而提高这个冷诱导启动子的活性。(C)2015爱思唯尔爱尔兰有限公司。保留所有权利。
Promoter activity is crucial for precise gene expression. Previously, a synthetic tuber-specific and cold-inducible promoter, pCL, containing a C-repeat/dehydration-responsive element (CRT/DRE) cassette and a tuber-specific fragment, was constructed in order to regulate cold-induced sweetening (CIS) in potatoes. However, the utility of pCL is limited due to its low activity. To improve its inducibility in response to low temperatures, we modified the CRT/DRE and flanking sequences. In particular, promoter activity was significantly improved by site-specific mutation of flanking sequences next to the core element (CCGAC) of CRT/DRE. We also inserted a modified CRT/DRE cassette into pCL; although this enhanced activity, it was not more effective than mutation of the flanking sequences. Indeed, up to 20-fold enhanced pCL activity could be achieved by replacing the CRT/DRE cassette in pCL with tandem repeats of two mutated CRT/DRE cassettes. This improvement was due to an enhanced affinity between the CRT/DRE cassette(s) and the StCBF1 transcription factor. Together, these data suggest that altering the structure of CRT/DRE can enhance CBF-related transcription complex formation and thus improve the activity of this cold-inducible promoter. (C) 2015 Elsevier Ireland Ltd. All rights reserved.