Novel methods for cloning and engineering genes using the polymerase chain reaction
Novel methods for cloning and engineering genes using the polymerase chain reaction
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DOI：
10.1016/0958-1669(95)80006-9
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发表时间：
1995-01-01
影响因子：
7.7
通讯作者：
Rashtchian, Ayoub
中科院分区：
文献类型：
--
作者：
Rashtchian, Ayoub
聚合酶链式反应 (PCR) 的使用在分子生物学的几乎所有方面都变得越来越广泛。最近，已经开发出新的不依赖于连接的方法来克隆使用 PCR 扩增的 DNA 片段。利用尿嘧啶 DNA 糖基化酶进行不依赖于连接的克隆（称为 UDG 克隆）为基因克隆和重组 PCR 提供了一种有效的方法。该技术现已应用于定点诱变、嵌套缺失的生成以及新型基因构建体的工程设计。这种方法的简便性和灵活性与 PCR 扩增相结合，简化了基因克隆和工程技术。
Use of the polymerase chain reaction (PCR) has become increasingly widespread in virtually all aspects of molecular biology. Recently, novel ligation-independent methods have been developed for the cloning of DNA fragments amplified using PCR. Ligation-independent cloning utilizing the enzyme uracil DNA glycosylase (termed UDG cloning) provides an efficient method for gene cloning and recombinant PCR. This technology is now being applied to site-directed mutagenesis, the generation of nested deletions, and the engineering of novel gene constructs. The ease and flexibility of this methodology, combined with PCR amplification, simplify gene cloning and engineering techniques.