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[天蓝色链霉菌A3(2)肌动噬菌体的物理图谱。VI. 利用肌动噬菌体φC31的缺失突变体构建噬菌体载体]

[Physical mapping of actinophage Streptomyces coelicolor A3(2). VI. The use of deletion mutants of actinophage phi C31 for construction of phage vectors].

作者信息

Sladkova I A, Klochkova O A, Lomovskaia N D

出版信息

Mol Biol (Mosk). 1982 Jul-Aug;16(4):739-44.

PMID:6289084
Abstract

Nonessential region responsible for G function has been identified in theta C31 phage genome by means of deletion mutants. The mutant phenotype is expressed upon theta C31 phage propagation in Streptomyces albus G strains differing in functioning of restriction and modification systems. Based on their increased resistance to EDTA, deletions were located in theta C31 delta 10 and theta C31 delta 65 phage mutants. Data are presented on physical mapping of nonessention region of theta C31 phage. The total length of this region is 24.1% of the overall length of DNA molecules. The DNA segment of 19.1% of the whole genome contains overlapped deletions. Theta C31 actinophage is proposed to be used as a cloning vector for Streptomyces. Various deletion mutants obtained, with the capacity of about 3 thousands base pairs may serve as "insertion vectors". The presence of the stretched nonessential genome region allows to use theta C31 phage as a "replacement vector". Then, insertion of foreign DNA to replace the EcoRI--C fragment of theta C31 DNA of 6.4 x 10(3) base pairs is possible. The phages comprising hybrid molecules may be selected for G and Lyg phenotypes.

摘要

通过缺失突变体已在θC31噬菌体基因组中鉴定出负责G功能的非必需区域。当θC31噬菌体在限制和修饰系统功能不同的白色链霉菌G菌株中繁殖时,突变表型得以表达。基于它们对EDTA的抗性增强,缺失位于θC31δ10和θC31δ65噬菌体突变体中。给出了关于θC31噬菌体非必需区域物理图谱的数据。该区域的总长度占DNA分子全长的24.1%。整个基因组19.1%的DNA片段包含重叠缺失。提议将θC31肌动噬菌体用作链霉菌的克隆载体。获得的各种缺失突变体,容量约为3千个碱基对,可作为“插入载体”。延伸的非必需基因组区域的存在使得可以将θC31噬菌体用作“置换载体”。然后,有可能插入外源DNA以取代6.4×10³碱基对的θC31 DNA的EcoRI - C片段。包含杂合分子的噬菌体可针对G和Lyg表型进行选择。

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