Analysis of the piggyBac transposase reveals a functional nuclear targeting signal in the 94 c-terminal residues.
Analysis of the piggyBac transposase reveals a functional nuclear targeting signal in the 94 c-terminal residues.
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DOI：
10.1186/1471-2199-9-72
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发表时间：
2008-08-11
影响因子：
--
通讯作者：
Fraser MJ Jr
中科院分区：
文献类型：
--
作者：
Keith JH;Fraser TS;Fraser MJ Jr
The piggyBac transposable element is a popular tool for germ-line transgenesis of eukaryotes. Despite this, little is known about the mechanism of transposition or the transposase (TPase) itself. A thorough understanding of just how piggyBac works may lead to more effective use of this important mobile element. A PSORTII analysis of the TPase amino acid sequence predicts a bipartite nuclear localization signal (NLS) near the c-terminus, just upstream of a putative ZnF (ZnF). We fused the piggyBac TPase upstream of and in-frame with the enhanced yellow fluorescent protein (EYFP) in the Drosophila melanogaster inducible metallothionein protein. Using Drosophila Schneider 2 (S2) cells and the deep red fluorescent nuclear stain Draq5, we were able to track the pattern of piggyBac localization with a scanning confocal microscope 48 hours after induction with copper sulphate. Through n and c-terminal truncations, targeted internal deletions, and specific amino acid mutations of the piggyBac TPase open reading frame, we found that not only is the PSORTII-predicted NLS required for the TPase to enter the nucleus of S2 cells, but there are additional requirements for negatively charged amino acids a short length upstream of this region for nuclear localization.
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DOI：
10.1073/pnas.0504679102
发表时间：
2005-11-08
影响因子：
11.1
作者：
Balu, B;Shoue, DA;Adams, JH
通讯作者：
Adams, JH
影响因子：
7.2
作者：
BOEHM, U;HEINLEIN, M;KUNZE, R
通讯作者：
KUNZE, R
影响因子：
64.5
作者：
Ding, S;Wu, XH;Xu, T
通讯作者：
Xu, T
DOI：
10.1073/pnas.95.13.7520
发表时间：
1998-06-23
影响因子：
11.1
作者：
Handler, AM;McCombs, SD;Saul, SH
通讯作者：
Saul, SH
影响因子：
2.6
作者：
Grossman, GL;Rafferty, CS;Benedict, MQ
通讯作者：
Benedict, MQ