Nishihara Hidenori, Okada Norihiro
Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Yokohama, Japan.
Methods Mol Biol. 2008;422:201-25. doi: 10.1007/978-1-59745-581-7_13.
Retroposons such as short interspersed elements (SINEs) and long interspersed elements are abundant transposable elements in eukaryote genomes. Recent large-scale comparative genome analyses have revealed that retroposons are a major component of genomes, wherein they provide structural diversity between species and uniqueness to each species. SINEs have been used as powerful markers in phylogenetic analyses of various species. This approach, which has been termed the SINE insertion method, infers phylogenetic relationships based on the presence/absence of SINEs among lineages. However, the method is not yet used extensively among biologists, especially molecular phylogenetists, because it is based on an understanding of the molecular mechanisms of retroposition, which may be unfamiliar to many researchers. Moreover, the method may require a large amount of bench work to characterize a new SINE family and to screen genomic libraries of the species of interest. In this chapter, we present the basic theory and detailed technical steps involved in a SINE insertion analysis. Furthermore, we explain the isolation and characterization of a new SINE family from the genome of a species of interest using as an example a known SINE family in mammals.
短散在元件(SINEs)和长散在元件等反转录转座子是真核生物基因组中丰富的可移动元件。最近的大规模比较基因组分析表明,反转录转座子是基因组的主要组成部分,它们在物种间提供结构多样性,并赋予每个物种独特性。SINEs已被用作各种物种系统发育分析中的有力标记。这种方法被称为SINE插入法,它根据谱系中SINEs的存在与否来推断系统发育关系。然而,这种方法在生物学家,尤其是分子系统发育学家中尚未得到广泛应用,因为它基于对反转录分子机制的理解,而许多研究人员可能对此并不熟悉。此外,该方法可能需要大量的实验工作来鉴定一个新的SINE家族,并筛选感兴趣物种的基因组文库。在本章中,我们介绍了SINE插入分析所涉及的基本理论和详细技术步骤。此外,我们以哺乳动物中一个已知的SINE家族为例,解释了从感兴趣物种的基因组中分离和鉴定一个新的SINE家族的过程。