Split-Cre complementation restores combination activity on transgene excision in hair roots of transgenic tobacco.
Split-Cre complementation restores combination activity on transgene excision in hair roots of transgenic tobacco.
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Split-Cre 互补恢复转基因烟草毛根中转基因切除的组合活性
DOI：
10.1371/journal.pone.0110290
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Luo K
中科院分区：
文献类型：
--
作者：
Wen M;Gao Y;Wang L;Ran L;Li J;Luo K
Cre/loxP系统越来越多地用于体外和体内DNA的遗传操作。以前有报道，当两个不同的启动子控制重叠共表达时，无活性的“split-Cre”片段可以恢复转基因小鼠中的Cre活性。在这项研究中，我们分析了分裂Cre蛋白的重组活性，发现没有重组酶活性检测在体外重组反应中，其中只有N-末端结构域（NCre）的分裂Cre蛋白的表达，而重组活性时，获得的C-末端（CCre）或NCre和CCre片段。我们还测定了在转基因烟草的发根中共表达的split-Cre蛋白的重组效率。当NCre或CCre基因单独表达时，在转基因烟草的发根中没有观察到Cre重组事件。与此相反，一个有效的重组事件被发现在转基因毛状根共表达两个非活性分裂Cre基因。此外，在转基因株系中，与核定位序列（NLS）融合的分裂Cre蛋白的恢复重组效率高于完整的Cre蛋白。因此，由split-Cre蛋白介导的DNA重组为转基因植物中基因表达的时空调控提供了另一种方法。
The Cre/loxP system is increasingly exploited for genetic manipulation of DNA in vitro and in vivo. It was previously reported that inactive ‘‘split-Cre’’ fragments could restore Cre activity in transgenic mice when overlapping co-expression was controlled by two different promoters. In this study, we analyzed recombination activities of split-Cre proteins, and found that no recombinase activity was detected in the in vitro recombination reaction in which only the N-terminal domain (NCre) of split-Cre protein was expressed, whereas recombination activity was obtained when the C-terminal (CCre) or both NCre and CCre fragments were supplied. We have also determined the recombination efficiency of split-Cre proteins which were co-expressed in hair roots of transgenic tobacco. No Cre recombination event was observed in hair roots of transgenic tobacco when the NCre or CCre genes were expressed alone. In contrast, an efficient recombination event was found in transgenic hairy roots co-expressing both inactive split-Cre genes. Moreover, the restored recombination efficiency of split-Cre proteins fused with the nuclear localization sequence (NLS) was higher than that of intact Cre in transgenic lines. Thus, DNA recombination mediated by split-Cre proteins provides an alternative method for spatial and temporal regulation of gene expression in transgenic plants.
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裂解2：通过分裂蛋白片段互补介导的DNA重组的时间控制。
影响因子：
3.7
作者：
Hirrlinger J;Requardt RP;Winkler U;Wilhelm F;Schulze C;Hirrlinger PG
通讯作者：
Hirrlinger PG
影响因子：
64.5
作者：
GU, W;SCHNEIDER, JW;NADALGINARD, B
通讯作者：
NADALGINARD, B
影响因子：
5.6
作者：
HOESS, R;ABREMSKI, K;MACK, A
通讯作者：
MACK, A
影响因子：
14.9
作者：
Jullien, N;Sampieri, F;Herman, JP
通讯作者：
Herman, JP
影响因子：
48
作者：
Miller, Ashleigh J.;Dudley, Sandra D.;Liskay, R. Michael
通讯作者：
Liskay, R. Michael