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转座酶是Tc1体外转座所需的唯一线虫蛋白。

Transposase is the only nematode protein required for in vitro transposition of Tc1.

作者信息

Vos J C, De Baere I, Plasterk R H

机构信息

The Netherlands Cancer Institute, Division of Molecular Biology, Amsterdam.

出版信息

Genes Dev. 1996 Mar 15;10(6):755-61. doi: 10.1101/gad.10.6.755.

Abstract

The Tc1 element of Caenorhabditis elegans is a member of the most widespread class of DNA transposons known in nature. Here, we describe efficient and precise transposition of Tc1 in a cell-free system. Tc1 appears to jump by a cut-and-paste mechanism of transposition. The terminal 26 bp of the Tc1 terminal repeats together with the flanking TA sequence are sufficient for transposition. The target site choice in vitro is similar to that in vivo. Transposition is achieved with an extract prepared from nuclei of transgenic nematodes that overexpress Tc1 transposase but also by recombinant transposase purified from Escherichia coli. The simple reaction requirements explain why horizontal spread of Tc1/mariner transposons can occur. They also suggest that Tcl may be a good vector for transgenesis of diverse animal species.

摘要

秀丽隐杆线虫的Tc1元件是自然界中已知分布最广泛的一类DNA转座子的成员。在此,我们描述了Tc1在无细胞系统中的高效且精确的转座。Tc1似乎通过一种剪切粘贴的转座机制进行跳跃。Tc1末端重复序列的末端26个碱基对与侧翼TA序列一起对于转座是足够的。体外的靶位点选择与体内相似。转座可通过从过表达Tc1转座酶的转基因线虫的细胞核制备的提取物实现,也可通过从大肠杆菌纯化的重组转座酶实现。简单的反应条件解释了Tc1/水手转座子为何能发生水平传播。它们还表明Tc1可能是用于多种动物物种转基因的良好载体。

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