Use of the gateway system for protein expression in multiple hosts.
Use of the gateway system for protein expression in multiple hosts.
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DOI：
10.1002/0471140864.ps0517s30
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发表时间：
2003-02-01
影响因子：
--
通讯作者：
Hartley, James L
中科院分区：
文献类型：
--
作者：
Hartley, James L
Gateway克隆方法允许克隆基因，随后通过体外位点特异性重组将其转移到任何载体中。然而，这并不一定意味着一个基因可以克隆一次，并在所有可用的宿主-载体组合中表达，得到一致满意的结果。这是因为不同的生物体具有不同的将mRNA翻译成蛋白质的机制，并且还因为在设计表达构建体时总是必须做出选择，例如存在或不存在终止密码子。本单元回顾了Gateway克隆，总结了限制在许多载体和宿主中使用一个克隆的普遍性的蛋白质表达方面，并讨论了如何最小化或解决基因的Gateway克隆的结构和蛋白质表达规则之间的冲突。Gateway克隆方法允许基因被克隆，随后通过体外位点特异性转移到任何载体中。
The Gateway cloning method allows a gene to be cloned and subsequently transferred into any vector by in vitro site-specific recombination. It does not necessarily follow, however, that a gene can be cloned once and expressed in all the available host-vector combinations with uniformly satisfactory results. This is because different organisms have different mechanisms of translating mRNA into protein, and also because choices always have to be made when designing an expression construct, for example, the presence or absence of a stop codon. This unit reviews Gateway cloning, summarizes aspects of protein expression that limit the universality of the use of one clone in many vectors and hosts, and discusses how conflicts between the structure of a Gateway clone of a gene and the rules of protein expression can be minimized or resolved.The Gateway cloning method allows a gene to be cloned and subsequently transferred into any vector by in vitro site-specific.