α complementation in the Cre recombinase enzyme
α complementation in the Cre recombinase enzyme
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DOI：
10.1002/gene.10227
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发表时间：
2003-09-01
期刊：
影响因子：
1.5
通讯作者：
Schütz, G
中科院分区：
文献类型：
--
作者：
Casanova, E;Lemberger, T;Schütz, G
Cre-IoxP系统越来越多地用于空间和时间基因失活。在这里，我们提出了一种新的方法来实现这一目标的选择性基因失活。根据β -半乳糖苷酶的互补模型，该酶被分裂成能够结合并维持酶活性的独立多肽，我们将Cre重组酶分为两个独立的多肽（一个包含NH2末端（α），另一个包含cooh末端（β））。单独来看，这两种多肽没有可检测到的活性。然而，当共表达时，多肽能够结合，产生Cre酶活性，其在体外最高可达野生型Cre重组酶的30%。我们提出了这种策略作为传统Cre-IoxP系统的修改，该系统可以通过在单独启动子的控制下表达两半来获得高度特异性的重组模式。(C) 2003 Wiley-Liss, Inc。
The Cre-IoxP system is increasingly exploited for spatial and temporal gene inactivation. Here we present a novel approach to achieve this goal of selective gene inactivation. Following the model of complementation in the beta-galactosidase enzyme, where the enzyme is split into independent polypeptides which are able to associate and maintain the enzymatic activity, we divided the Cre recombinase into two independent polypeptides (one containing the NH2 terminus (alpha) and a second one containing the COOH-terminus (beta)). Individually, the two polypeptides have no detectable activity. However, when coexpressed the polypeptides are able to associate, giving rise to Cre enzymatic activity, which optimally is as high as 30% of that seen with wildtype Cre recombinase in vitro. We present this strategy as a modification of the traditional Cre-IoxP system, which could be used to obtain a highly specific recombination pattern by expressing the two halves under the control of separate promoters. (C) 2003 Wiley-Liss, Inc.