PCR检测结果表明,在非洲爪蟾(Xenopus laevis)中,medaka fish Tol1转座因子具有可移动性。

Akira Hikosaka, Akihiko Koga
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引用次数: 5

摘要

Tol1是medaka鱼类Oryzias latipes中发现的基于dna的转座元件,是hAT (hobo/Activator/Tam3)转座元件家族的成员。除了最初的宿主物种外,它的移动性已经在人类和老鼠身上得到了证明。因此,该元件有望在广泛的脊椎动物中作为基因组操作工具而发挥作用。本研究表明,非洲爪蟾(Xenopus laevis)是脊椎动物遗传学和发育生物学的主要模式生物,其Tol1基因可以被切除。将携带Tol1元件的指示质粒与编码全长Tol1转座酶的辅助质粒或产生截断蛋白的修饰的辅助质粒一起注射到2或4细胞期胚胎中,从尾期胚胎中回收。在全长转座酶的实验中观察到指示质粒的Tol1区域缺失,而在其他情况下没有。这种缺失与断点上的各种足迹序列有关,正如在许多基于dna的转座元件中经常观察到的那样。这些结果表明,在转座酶的催化下,Tol1元件从指示质粒中被切除,表明Tol1元件在该蛙种中是可移动的。
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PCR detection of excision suggests mobility of the medaka fish Tol1 transposable element in the frog Xenopus laevis.

Tol1 is a DNA-based transposable element identified in the medaka fish Oryzias latipes and a member of the hAT (hobo/Activator/Tam3) transposable element family. Its mobility has already been demonstrated in the human and mouse, in addition to its original host species. This element is thus expected to be useful in a wide range of vertebrates as a genomic manipulation tool. Herein, we show that the Tol1 element can undergo excision in the African clawed frog Xenopus laevis, a major model organism for vertebrate genetics and developmental biology. An indicator plasmid carrying a Tol1 element was injected into 2- or 4-cell-stage embryos together with either a helper plasmid coding for the full-length Tol1 transposase or a modified helper plasmid yielding a truncated protein, and recovered from tailbud-stage embryos. Deletion of the Tol1 region of the indicator plasmid was observed in the experiment with the full-length transposase, and not in the other case. The deletion was associated with various footprint sequences at breakpoints, as frequently observed with many DNA-based transposable elements. These results indicate that the Tol1 element was excised from the indicator plasmid by catalysis of the transposase, and suggest that the Tol1 element is mobile in this frog species.

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