北极七鳃鳗染色体位点特异性重复序列的分子细胞遗传学特征

J. Ishijima, Y. Uno, Mitsuo Nunome, C. Nishida, Shigehiro Kuraku, Y. Matsuda
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引用次数: 10

摘要

摘要现存的七鳃鳗核型几乎都具有点状微染色体的特征。为了了解七鳃鳗染色体结构的分子基础,我们对北极七鳃鳗(Lethenteron camtschaticum)的18S-28S和5S核糖体RNA (rRNA)基因和端粒TTAGGG重复序列进行了染色体c带和银染色和染色体定位。此外,我们克隆了染色体特定位点的重复DNA序列,并通过核苷酸测序、染色体原位杂交和过滤杂交对其进行了表征。检测到三种类型的重复序列;一个富含200 bp at的重复序列,LCA-EcoRIa,与3对染色体的18S-28S rRNA基因簇共定位;一个364 bp的富含at的LCA-EcoRIb序列与海七鳃鳗(Petromyzon marinus)的EcoRI序列家族具有同源性,该序列包含短重复序列作为着丝性基序;一个富含gc的702 bp LCA-ApaI序列分布在几乎所有染色体上,与Ty3/Gypsy家族长末端重复(LTR)反转录转座子的整合酶编码区具有显著的同源性。所有这三个重复序列在石鳖科或石鳖科和石鳖科中都高度保守。这些位点特异性重复序列的分子细胞遗传学特征表明,它们可能与程序性基因组重排(LCA-EcoRIa)、着丝粒结构和功能(LCA-EcoRIb)以及通过非同源染色体间均质化对LTR逆转录元件的位点特异性扩增(LCA-ApaI)有关。
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Molecular cytogenetic characterization of chromosome site-specific repetitive sequences in the Arctic lamprey (Lethenteron camtschaticum, Petromyzontidae)
Abstract All extant lamprey karyotypes are characterized by almost all dot-shaped microchromosomes. To understand the molecular basis of chromosome structure in lampreys, we performed chromosome C-banding and silver staining and chromosome mapping of the 18S–28S and 5S ribosomal RNA (rRNA) genes and telomeric TTAGGG repeats in the Arctic lamprey (Lethenteron camtschaticum). In addition, we cloned chromosome site-specific repetitive DNA sequences and characterized them by nucleotide sequencing, chromosome in situ hybridization, and filter hybridization. Three types of repetitive sequences were detected; a 200-bp AT-rich repetitive sequence, LCA-EcoRIa that co-localized with the 18S–28S rRNA gene clusters of 3 chromosomal pairs; a 364-bp AT-rich LCA-EcoRIb sequence that showed homology to the EcoRI sequence family from the sea lamprey (Petromyzon marinus), which contains short repeats as centromeric motifs; and a GC-rich 702-bp LCA-ApaI sequence that was distributed on nearly all chromosomes and showed significant homology with the integrase-coding region of a Ty3/Gypsy family long terminal repeat (LTR) retrotransposon. All three repetitive sequences are highly conserved within the Petromyzontidae or within Petromyzontidae and Mordaciidae. Molecular cytogenetic characterization of these site-specific repeats showed that they may be correlated with programed genome rearrangement (LCA-EcoRIa), centromere structure and function (LCA-EcoRIb), and site-specific amplification of LTR retroelements through homogenization between non-homologous chromosomes (LCA-ApaI).
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