van Luenen H G, Colloms S D, Plasterk R H
Netherlands Cancer Institute, Division of Molecular Biology, Amsterdam.
EMBO J. 1993 Jun;12(6):2513-20. doi: 10.1002/j.1460-2075.1993.tb05906.x.
The commonly studied Caenorhabditis elegans strain Bristol N2 contains approximately 15 copies per genome of the transposon Tc3. However, Tc3 is not active in Bristol N2. Tc3 contains one major open reading frame (Tc3A). We have fused this open reading frame to an inducible promoter and expressed it in a transgenic Bristol N2 line. Tc3A expression resulted in frequent excision and transposition of endogenous Tc3 elements. This shows that the Bristol N2 genome contains Tc3 transposons that are cis proficient for transposition, but are immobile because Tc3A is absent. We demonstrate that recombinant Tc3A binds specifically to the terminal nucleotides of the Tc3 inverted repeat, indicating that Tc3A is the Tc3 transposase. Activation of Tc3 transposition in vivo was accompanied by the appearance of extrachromosomal, linear copies of Tc3. These may be intermediates in Tc3 transposition.
常用的秀丽隐杆线虫布里斯托N2品系每个基因组中约含有15个转座子Tc3拷贝。然而,Tc3在布里斯托N2中不活跃。Tc3含有一个主要的开放阅读框(Tc3A)。我们已将此开放阅读框与一个可诱导启动子融合,并在一个转基因布里斯托N2品系中进行表达。Tc3A的表达导致内源性Tc3元件频繁切除和转座。这表明布里斯托N2基因组含有对转座顺式作用 proficient的Tc3转座子,但由于缺乏Tc3A而不能移动。我们证明重组Tc3A特异性结合Tc3反向重复序列的末端核苷酸,表明Tc3A是Tc3转座酶。体内Tc3转座的激活伴随着染色体外线性Tc3拷贝的出现。这些可能是Tc3转座的中间体。