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番茄功能基因组学的增强子捕获插入突变体集。

A collection of enhancer trap insertional mutants for functional genomics in tomato.

机构信息

Centro de Investigación en Biotecnología Agroalimentaria (BITAL), Universidad de Almería, Almería, Spain.

Instituto de Biología Molecular y Celular de Plantas (UPV-CSIC), Universidad Politécnica de Valencia, Valencia, Spain.

出版信息

Plant Biotechnol J. 2017 Nov;15(11):1439-1452. doi: 10.1111/pbi.12728. Epub 2017 Apr 20.

Abstract

With the completion of genome sequencing projects, the next challenge is to close the gap between gene annotation and gene functional assignment. Genomic tools to identify gene functions are based on the analysis of phenotypic variations between a wild type and its mutant; hence, mutant collections are a valuable resource. In this sense, T-DNA collections allow for an easy and straightforward identification of the tagged gene, serving as the basis of both forward and reverse genetic strategies. This study reports on the phenotypic and molecular characterization of an enhancer trap T-DNA collection in tomato (Solanum lycopersicum L.), which has been produced by Agrobacterium-mediated transformation using a binary vector bearing a minimal promoter fused to the uidA reporter gene. Two genes have been isolated from different T-DNA mutants, one of these genes codes for a UTP-glucose-1-phosphate uridylyltransferase involved in programmed cell death and leaf development, which means a novel gene function reported in tomato. Together, our results support that enhancer trapping is a powerful tool to identify novel genes and regulatory elements in tomato and that this T-DNA mutant collection represents a highly valuable resource for functional analyses in this fleshy-fruited model species.

摘要

随着基因组测序项目的完成,下一个挑战是缩小基因注释和基因功能分配之间的差距。用于识别基因功能的基因组工具基于对野生型和突变体之间表型变异的分析;因此,突变体库是一种有价值的资源。从这个意义上说,T-DNA 库允许轻松、直接地鉴定标记基因,是正向和反向遗传策略的基础。本研究报告了番茄(Solanum lycopersicum L.)增强子捕获 T-DNA 库的表型和分子特征,该库是通过农杆菌介导的转化用带有最小启动子融合 uidA 报告基因的二元载体产生的。从不同的 T-DNA 突变体中分离出了两个基因,其中一个基因编码参与程序性细胞死亡和叶片发育的 UTP-葡萄糖-1-磷酸尿苷酰转移酶,这意味着在番茄中报告了一个新的基因功能。总之,我们的结果支持增强子捕获是鉴定番茄中新基因和调控元件的有力工具,并且该 T-DNA 突变体库代表了肉质果实模式物种功能分析的极有价值的资源。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/77ab/11389220/b6de2a8e6bcf/PBI-15-1439-g005.jpg

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