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首页> 外文期刊>Molecular biology and evolution >Microuli, a family of miniature subterminal inverted-repeat transposable elements (MSITEs): Transposition without terminal inverted repeats
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Microuli, a family of miniature subterminal inverted-repeat transposable elements (MSITEs): Transposition without terminal inverted repeats

机译：Microuli，微型亚末端反向重复转座因子（MSITE）系列：转座而无末端反向重复

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The first Microuli element, Microuli-Aal, was discovered as a 209-bp insertion in the 5' long terminal repeat of a retrotransposon named Mosqcopia-Aa2 (unpublished data, GenBank AY009101) in the yellow fever mosquito, Aedes aegypti. As shown in figure lA, the Microuli insertion resulted in a TTAA target duplication, indicating past mobility of the element. To further characterize this mobile element, we screened an A. aegypti genomic library, using a digoxigenin-labeled singlestranded DNA probe generated by asymmetric PCR using the entire Microuli-Aal as the template. The library construction, probe labeling, and library screening procedures were as previously described (Tu 2000). The final washing stringency was 0.5 X SSC with 0.1 percent SDS at 55 deg C, which allows approximately 15 percent-25 percent mismatch (Meinkoth and Wahl 1984). Because 462 positive plaques were identified among 64,000 total plaques screened, it was estimated using a previously described calculation method (Tu 2000) that there were approximately 3,000 copies of Microuli per haploid genome. The entire inserts of three positive genomic clones isolated during the above screening experiment were sequenced and deposited in GenBank (AY009102-AY009104). The boundaries of Microuli elements were deduced based on sequence comparisons between the four genomic clones. As shown in figure lB, three of the four elements have the same termini and are flanked by putative TTAA target duplications. There were no sequence similarities between these Microuli clones out-side the TTAA target duplications. Although we cannot determine the extent of the diversity of Microuli elements based on the analysis of four elements, the evidence of insertion, sequence and size homogeneity between three out of the four copies, and the conservation of putative target duplications suggest that Microuli has been an intact unit of transposition which has generated a large number of copies.
机译：在黄热伊蚊埃及伊蚊中，第一个Microuli元件Microuli-Aal被发现为名为Mosqcopia-Aa2的反转录转座子5'长末端重复序列中209 bp的插入（未公开数据，GenBank AY009101）。如图1A所示，Microuli插入导致TTAA靶标重复，表明该元件过去的迁移性。为了进一步表征该可移动元件，我们使用洋地黄毒苷标记的单链DNA探针（通过使用整个Microuli-Aal作为模板的不对称PCR产生）筛选了埃及埃及曲霉基因组文库。文库的构建，探针标记和文库筛选程序如前所述（Tu 2000）。最终的洗涤严格度为0.5 X SSC，在55摄氏度下具有0.1％SDS，这允许大约15％-25％的不匹配（Meinkoth和Wahl 1984）。因为在所筛选的64,000个总噬菌斑中鉴定出462个阳性噬菌斑，所以使用先前描述的计算方法（Tu 2000）估计每个单倍体基因组大约有3,000个微粒。对在上述筛选实验中分离出的三个阳性基因组克隆的整个插入片段进行测序，并保存在GenBank（AY009102-AY009104）中。基于四个基因组克隆之间的序列比较推导了Microuli元素的边界。如图1B所示，四个元件中的三个具有相同的末端，并且两侧是假定的TTAA靶标重复。在TTAA靶标重复之外，这些Microuli克隆之间没有序列相似性。尽管我们无法通过对四个元素的分析来确定微菌元素的多样性程度，但四拷贝中三个拷贝之间的插入，序列和大小均一性的证据以及推定的靶标重复的保守性表明，微菌一直是一种完整的换位单位，已产生大量拷贝。

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