通过热不对称交错PCR（TAIL-PCR）对T-DNA和Ac/Ds进行定位以分析染色体重排

Mapping of T-DNA and Ac/Ds by TAIL-PCR to Analyze Chromosomal Rearrangements.

作者信息

Fujimoto Satoru, Matsunaga Sachihiro, Murata Minoru

机构信息

Institute of Plant Science and Resources, Okayama University, 2-20-1, Chuo, Kurashiki, 710-0046, Japan.

Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science, 2641, Yamazaki, Noda, Chiba, 278-8510, Japan.

出版信息

Methods Mol Biol. 2016;1469:207-16. doi: 10.1007/978-1-4939-4931-1_17.

DOI:10.1007/978-1-4939-4931-1_17

PMID:27557698

Abstract

Insertion mutagenesis using known DNA sequences such as T-DNA and transposons is an important tool for studies on gene function in plant sciences. The transposons Activator (Ac)/Dissociation (Ds) have been systematically used to manipulate plant chromosomes. For both of these applications, the recovery of genomic DNA sequences flanking the insertions is required to estimate the sizes and/or scales of the reconstituted chromosomes. In this chapter, we describe the protocols for thermal asymmetric interlaced PCR (TAIL-PCR) for isolation of genomic sequences flanking DNA inserts in plant genomes.

摘要

利用已知DNA序列（如T-DNA和转座子）进行插入诱变是植物科学中研究基因功能的重要工具。转座子激活子（Ac）/解离子（Ds）已被系统地用于操纵植物染色体。对于这两种应用，都需要回收插入位点两侧的基因组DNA序列，以估计重组染色体的大小和/或规模。在本章中，我们描述了用于分离植物基因组中DNA插入片段两侧基因组序列的热不对称交错PCR（TAIL-PCR）方法。