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Ligand-regulated site-specific recombination.

机译:配体调节的位点特异性重组。

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摘要

Site-specific recombination offers a potential way to alter a living genome by design in a precise and stable manner. This potential requires strategies which can be used to regulate the recombination event. We describe a strategy to regulate FLP recombinase activity which relies on expressing FLP as a fusion protein with steroid hormone receptor ligand binding domains (LBDs). In the absence of a ligand cognate to the LBD, the recombinase activity of the fusion protein is extremely low. Upon ligand administration, recombinase activity is rapidly induced. These results outline the basis for inducible expression or disruption strategies based on inducible recombination. Additionally, we have exploited the conditional nature of FLP-LBD fusion proteins to direct integration of a plasmid into a specific genomic site at frequencies approaching the frequency of random integration.
机译:位点特异性重组提供了一种潜在的方式,可通过精确,稳定的设计来改变活的基因组。这种潜力需要可用于调节重组事件的策略。我们描述了一种调节FLP重组酶活性的策略,该策略依赖于表达FLP为与类固醇激素受体配体结合域(LBDs)的融合蛋白。在不存在与LBD同源的配体的情况下,融合蛋白的重组酶活性极低。配体给药后,重组酶活性被迅速诱导。这些结果概述了基于可诱导重组的可诱导表达或破坏策略的基础。此外,我们已经利用FLP-LBD融合蛋白的条件性质,以接近随机整合频率的频率将质粒整合到特定的基因组位点。

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