偶然分枝杆菌偶然变种质粒pAL5000的限制性内切酶图谱分析与克隆

Restriction endonuclease mapping and cloning of Mycobacterium fortuitum var. fortuitum plasmid pAL5000.

作者信息

Labidi A, David H L, Roulland-Dussoix D

出版信息

Ann Inst Pasteur Microbiol (1985). 1985 Sep-Oct;136B(2):209-15. doi: 10.1016/s0769-2609(85)80045-4.

Abstract

A restriction map of Mycobacterium fortuitum var. fortuitum plasmid pAL5000 was established. The unique sites for ApaI, BamHI, BglII, BstEII, ClaI, EcoRI, EcoRV, HpaI, KpnI and NarI were located on the 5.0-Kb plasmid. The plasmid had no sites for AhaIII, BclI, HindIII, PstI, SphI and XbaI. pAL5000 was cloned into pBR322 and propagated in Escherichia coli. Three hybrid pAL5000-pBR322 plasmids carrying the complete pAL5000 sequence were constructed by joining the plasmids at their BamHI, EcoRI or EcoRV sites. We also cloned into these plasmids a 1489-bp DNA fragment conferring resistance to kanamycin and originating from the streptococcal plasmid pJH1. The construction of these plasmids will facilitate the analysis and manipulation of pAL5000, and may allow the development of a vector system for genetic analysis in mycobacteria.

摘要

构建了偶然分枝杆菌偶然变种质粒pAL5000的限制性图谱。ApaI、BamHI、BglII、BstEII、ClaI、EcoRI、EcoRV、HpaI、KpnI和NarI的独特位点位于5.0-Kb的质粒上。该质粒没有AhaIII、BclI、HindIII、PstI、SphI和XbaI的位点。pAL5000被克隆到pBR322中并在大肠杆菌中繁殖。通过在其BamHI、EcoRI或EcoRV位点连接质粒，构建了三个携带完整pAL5000序列的杂交pAL5000-pBR322质粒。我们还将一个1489-bp的赋予卡那霉素抗性且来源于链球菌质粒pJH1的DNA片段克隆到这些质粒中。这些质粒的构建将有助于对pAL5000进行分析和操作，并可能有助于开发一种用于分枝杆菌遗传分析的载体系统。