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首页> 外文期刊>Molecular Plant >Illumina Sequencing Technology as a Method of Identifying T-DNA Insertion Loci in Activation-Tagged Arabidopsis thaliana Plants.
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Illumina Sequencing Technology as a Method of Identifying T-DNA Insertion Loci in Activation-Tagged Arabidopsis thaliana Plants.

机译：Illumina测序技术，作为鉴定激活标签拟南芥植物中T-DNA插入位点的方法。

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摘要

Forward genetic screens are commonly used as unbiased tools to isolate genes responsible for a phenotype of interest. In Arabidopsis thaliana, especially T-DNA activation tagging populations are frequently employed. These populations are generated using vectors containing multiple copies of the constitutive 35S promoters derived from cauliflower mosaic virus (35S CaMV) and often result in isolation of dominant gain-of-function alleles (Weigel et al., 2000; Nakazawa et al., 2003). This allows the study of members of large gene families that are often functionally redundant and, therefore, hard to identify in loss-of-function screens. Moreover, due to the dominant nature, the phenotypes can usually be recognized in T1 generations (Ostergaard and Yanofsky, 2004). Plasmid rescue and thermal asymmetric interlaced PCR (TAIL-PCR) have been effectively employed to recover plant-specific sequences flanking the T-DNA insertions (Weigel et al., 2000; Singer and Burke, 2003). However, in some instances, these two techniques do not yield expected results, probably due to potential sequence complexities following integration events (Laufs et al., 1999).
机译：正向遗传筛选通常用作无偏工具来分离引起目标表型的基因。在拟南芥中，尤其是经常使用T-DNA激活标签种群。这些种群是通过使用包含多拷贝自花椰菜花叶病毒（35S CaMV）的组成型35S启动子的载体产生的，通常会导致功能获得性优势等位基因的分离（Weigel等，2000; Nakazawa等，2003）。 ）。这允许研究通常在功能上是冗余的大型基因家族的成员，因此很难在功能丧失的筛选中进行鉴定。此外，由于具有支配性，这些表型通常可以在T1代中被识别（Ostergaard和Yanofsky，2004）。质粒拯救和热不对称交错PCR（TAIL-PCR）已被有效地用于回收T-DNA插入侧翼的植物特异性序列（Weigel等，2000; Singer和Burke，2003）。但是，在某些情况下，这两种技术可能无法产生预期的结果，这可能是由于整合事件后潜在的序列复杂性所致（Laufs等，1999）。

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