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质粒载体pBR322及其专用衍生物——综述

Plasmid vector pBR322 and its special-purpose derivatives--a review.

作者信息

Balbás P, Soberón X, Merino E, Zurita M, Lomeli H, Valle F, Flores N, Bolivar F

出版信息

Gene. 1986;50(1-3):3-40. doi: 10.1016/0378-1119(86)90307-0.

Abstract

The plasmid pBR322 was one of the first EK2 multipurpose cloning vectors to be designed and constructed (ten years ago) for the efficient cloning and selection of recombinant DNA molecules in Escherichia coli. This 4363-bp DNA molecule has been extensively used as a cloning vehicle because of its simplicity and the availability of its nucleotide sequence. The widespread use of pBR322 has prompted numerous studies into its molecular structure and function. These studies revealed two features that detract from the plasmid's effectiveness as a cloning vector: plasmid instability in the absence of selection and, the lack of a direct selection scheme for recombinant DNA molecules. Several vectors based on pBR322 have been constructed to overcome these limitations and to extend the vector's versatility to accommodate special cloning purposes. The objective of this review is to provide a survey of these derivative vectors and to summarize information currently available on pBR322.

摘要

质粒pBR322是最早(十年前)设计和构建的用于在大肠杆菌中高效克隆和筛选重组DNA分子的EK2多功能克隆载体之一。这个4363碱基对的DNA分子因其简单性和核苷酸序列的可获得性而被广泛用作克隆载体。pBR322的广泛使用促使人们对其分子结构和功能进行了大量研究。这些研究揭示了两个降低该质粒作为克隆载体有效性的特征:在无选择条件下质粒的不稳定性,以及缺乏针对重组DNA分子的直接筛选方案。基于pBR322构建了几种载体以克服这些限制,并扩展该载体的通用性以适应特殊的克隆目的。本综述的目的是对这些衍生载体进行概述,并总结目前关于pBR322的可用信息。

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