Gbadegesin M A, Beeching J R
Department of Biology and Biochemistry, University of Bath, Bath, United Kingdom.
Genet Mol Res. 2010 Apr 13;9(2):639-50. doi: 10.4238/vol9-2gmr713.
Transposable elements contribute to the size, structure, variation, and diversity of the genome and have major effects on gene function. Sequencing projects have revealed the diversity of transposable elements in many organisms and have shown that they constitute a high percentage of the genome. PCR-based techniques using degenerate primers designed from conserved enzyme domains of transposable elements can provide quick and extensive surveys, making study of diversity and abundance and their applications possible in species where full genome sequence data are not yet available. We studied cassava (Manihot esculenta) En/Spm-like transposons (Meens) with regard to genomic distribution, sequence diversity and methylation status. Cassava transposase fragments characteristic of En/Spm-like transposons were isolated, cloned and characterized. Sequence analysis showed that cassava En/Spm-like elements are highly conserved, with overall identity in the range of 68-98%. Southern hybridization supports the presence of multiple copies of En/Spm-like transposons integrated in the genome of all cassava cultivars that we tested. Hybridization patterns of HpaII- and MspI-digested cassava genomic DNA revealed highly methylated sequences. There were no clear differences in hybridization pattern between the cultivars. We did not detect RNA transcripts of Meens by Northern procedures. We examined the possibility of recent transposition activities of the cassava En/Spm-like elements.
转座元件影响基因组的大小、结构、变异和多样性,并对基因功能有重要作用。测序项目揭示了许多生物中转座元件的多样性,并表明它们在基因组中占很高比例。利用根据转座元件保守酶结构域设计的简并引物进行基于PCR的技术,可以提供快速而广泛的调查,从而使在尚未获得全基因组序列数据的物种中研究转座元件的多样性、丰度及其应用成为可能。我们研究了木薯(Manihot esculenta)中类En/Spm转座子(Meens)的基因组分布、序列多样性和甲基化状态。分离、克隆并鉴定了具有类En/Spm转座子特征的木薯转座酶片段。序列分析表明,木薯类En/Spm元件高度保守,总体一致性在68%-98%之间。Southern杂交支持在我们测试的所有木薯品种的基因组中存在多个整合的类En/Spm转座子拷贝。HpaII和MspI消化的木薯基因组DNA的杂交模式显示出高度甲基化的序列。各品种间杂交模式没有明显差异。我们通过Northern方法未检测到Meens的RNA转录本。我们研究了木薯类En/Spm元件近期转座活性的可能性。