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科莫多巨蜥的线粒体基因组:一种使用针对爬行动物的引物的高效测序方法及新的基因重排

Mitochondrial genome of the Komodo dragon: efficient sequencing method with reptile-oriented primers and novel gene rearrangements.

作者信息

Kumazawa Yoshinori, Endo Hideki

机构信息

Graduate School of Science, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8602, Japan.

出版信息

DNA Res. 2004 Apr 30;11(2):115-25. doi: 10.1093/dnares/11.2.115.

Abstract

The mitochondrial genome of the Komodo dragon (Varanus komodoensis) was nearly completely sequenced, except for two highly repetitive noncoding regions. An efficient sequencing method for squamate mitochondrial genomes was established by combining the long polymerase chain reaction (PCR) technology and a set of reptile-oriented primers designed for nested PCR amplifications. It was found that the mitochondrial genome had novel gene arrangements in which genes from NADH dehydrogenase subunit 6 to proline tRNA were extensively shuffled with duplicate control regions. These control regions had 99% sequence similarity over 700 bp. Although snake mitochondrial genomes are also known to possess duplicate control regions with nearly identical sequences, the location of the second control region suggested independent occurrence of the duplication on lineages leading to snakes and the Komodo dragon. Another feature of the mitochondrial genome of the Komodo dragon was the considerable number of tandem repeats, including sequences with a strong secondary structure, as a possible site for the slipped-strand mispairing in replication. These observations are consistent with hypotheses that tandem duplications via the slipped-strand mispairing may induce mitochondrial gene rearrangements and may serve to maintain similar copies of the control region.

摘要

科莫多巨蜥(Varanus komodoensis)的线粒体基因组除了两个高度重复的非编码区域外,几乎已被完全测序。通过结合长聚合酶链反应(PCR)技术和一组为巢式PCR扩增设计的面向爬行动物的引物,建立了一种用于有鳞目线粒体基因组的高效测序方法。研究发现,该线粒体基因组具有新的基因排列方式,其中从NADH脱氢酶亚基6到脯氨酸tRNA的基因与重复的控制区域广泛重排。这些控制区域在700 bp的长度上具有99%的序列相似性。尽管已知蛇的线粒体基因组也拥有序列几乎相同的重复控制区域,但第二个控制区域的位置表明,在导致蛇和科莫多巨蜥的谱系上,重复事件是独立发生的。科莫多巨蜥线粒体基因组的另一个特征是存在大量串联重复序列,包括具有强大二级结构的序列,这些序列可能是复制过程中滑链错配的位点。这些观察结果与以下假设一致:通过滑链错配的串联重复可能会诱导线粒体基因重排,并可能有助于维持控制区域的相似拷贝。

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