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Retrotransposable elements on the W chromosome of the silkworm, Bombyx mori

机译:家蚕W染色体上的可逆转座元件

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The sex chromosomes of the silkworm, Bombyx mori, are designated ZW(XY) for females and ZZ(XX) for males. The W chromosome of B. mori does not recombine with the Z chromosome and autosomes and no genes for morphological characters have been mapped to the W chromosome as yet. Furthermore, femaleness is determined by the presence of a single W chromosome, regardless of the number of autosomes or Z chromosomes. To understand these interesting features of the W chromosome, it is necessary to analyze the W chromosome at the molecular biology level. Initially to isolate DNA sequences specific for the W chromosome as randomly amplified polymorphic DNA (RAPD) markers, we compared the genomic DNAs between males and females by PCR with arbitrary 10-mer primers. To the present, we have identified 12 W-specific RAPD markers, and with the exception of one RAPD marker, all of the deduced amino acid sequences of these W-specific RAPD markers show similarity to previously reported amino acid sequences of retrotransposable elements from various organisms. After constructing a genomic DNA lambda phage library of B. mori we obtained two lambda phage clones, one containing the W-Kabuki RAPD sequence and one containing the W-Samurai RAPD sequence and found that these DNA sequences comprised nested structures of many retrotransposable elements. To further analyze the W chromosome, we obtained 14 W-specific bacterial artificial chromosome (BAC) clones from three BAC libraries and subjected these clones to shotgun sequencing. The resulting assembly of sequences did not produce a single contiguous sequence due to the presence of many retrotransposable elements. Therefore, we coupled PCR with shotgun sequencing. Through these analyses, we found that many long terminal repeat (LTR) and non-LTR retrotransposons, retroposons, DNA transposons and their derivatives, have accumulated on the W chromosome as strata. These results strongly indicate that retrotransposable elements are the main structural component of the W chromosome. Copyright (c) 2005 S. Karger AG, Basel.
机译:家蚕的性染色体,雌性命名为ZW(XY),雄性命名为ZZ(XX)。桑蚕的W染色体尚未与Z染色体和常染色体重组,并且尚未将形态学特征的基因定位到W染色体。此外,雌性由单个W染色体的存在决定,而与常染色体或Z染色体的数量无关。要了解W染色体的这些有趣特征,有必要在分子生物学水平上分析W染色体。最初,我们以随机扩增的多态性DNA(RAPD)标记来分离W染色体特有的DNA序列,我们通过PCR与任意10-mer引物比较了男性和女性之间的基因组DNA。到目前为止,我们已经鉴定出12种W特异性RAPD标记,除一个RAPD标记外,所有这些W特异性RAPD标记的推导氨基酸序列都与先前报道的来自各种不同方向的可逆转座子氨基酸序列相似生物。构建家蚕的基因组DNAλ噬菌体文库后,我们获得了两个λ噬菌体克隆,一个含有W-Kabuki RAPD序列,一个含有W-Samurai RAPD序列,发现这些DNA序列包含许多可逆转座子的嵌套结构。为了进一步分析W染色体,我们从三个BAC库中获得了14个W特异的细菌人工染色体(BAC)克隆,并对这些克隆进行了shot弹枪测序。由于存在许多可逆转座的元件,因此所得的序列组装未产生单个连续序列。因此,我们将PCR与散弹枪测序相结合。通过这些分析,我们发现许多长末端重复序列(LTR)和非LTR逆转座子,逆转座子,DNA转座子及其衍生物已经作为层在W染色体上积累。这些结果强烈表明,逆转座因子是W染色体的主要结构成分。版权所有(c)2005 S.Karger AG,巴塞尔。

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