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噬菌体LL-H介导的德氏乳杆菌中有效质粒pX3转导

Effective plasmid pX3 transduction in Lactobacillus delbrueckii by bacteriophage LL-H.

作者信息

Ravin Victor, Sasaki Takashi, Räisänen Liisa, Riipinen Katja-Anneli, Alatossava Tapani

机构信息

Department of Biology, University of Oulu, Linnanmaa campus, P.O. Box 3000, FIN-90014 Oulu, Finland.

出版信息

Plasmid. 2006 May;55(3):184-93. doi: 10.1016/j.plasmid.2005.12.003. Epub 2006 Feb 3.

Abstract

High-frequency plasmid transductions in Lactobacillus delbrueckii subsp. lactis and subsp. bulgaricus strains mediated by pac-type bacteriophages were observed and further investigated. The frequency of plasmid transduction by phages LL-H and LL-S attained levels of from 0.10 to about 1 with plasmid p X 3, but only about 2 x 10(-2) with plasmid pJK650. Infection of L. delbrueckii subsp. lactis strain LKT(pX3) or ATCC 15808(pX3) with phage LL-H resulted in intensive concatemerization of plasmid pX3, and most progeny phage particles contained concatemers of plasmid DNA instead of phage LL-H DNA. The synthesis of phage LL-H DNA was depressed. No evident homology or recombination was observed between phage LL-H DNA and plasmid pX3. The unusually high frequency of plasmid pX3 transduction by phage LL-H could be considered to result from specific interaction(s) between a particular phage and plasmid. These interactions may include pX3-mediated blockage of phage LL-H DNA replication and effective use of a particular pac-like site located about 1 kb from BglII in the smaller NdeI-BglII fragment of plasmid pX3. Phage LL-H together with plasmid vector pX3 could be used as effective plasmid transduction tools for genetic engineering of L. delbrueckii subsp. lactis and subsp. bulgaricus strains.

摘要

观察到并进一步研究了由pac型噬菌体介导的德氏乳杆菌乳酸亚种和保加利亚亚种菌株中的高频质粒转导。噬菌体LL-H和LL-S对质粒pX3的转导频率达到了0.10至约1的水平,但对质粒pJK650的转导频率仅约为2×10⁻²。用噬菌体LL-H感染德氏乳杆菌乳酸亚种菌株LKT(pX3)或ATCC 15808(pX3)导致质粒pX3强烈串联化,并且大多数子代噬菌体颗粒含有质粒DNA的串联体而非噬菌体LL-H DNA。噬菌体LL-H DNA的合成受到抑制。未观察到噬菌体LL-H DNA与质粒pX3之间有明显的同源性或重组。噬菌体LL-H对质粒pX3的异常高频率转导可被认为是由特定噬菌体与质粒之间的特异性相互作用导致的。这些相互作用可能包括pX3介导的对噬菌体LL-H DNA复制的阻断以及对位于质粒pX3较小的NdeI-BglII片段中距BglII约1 kb处的特定类pac位点的有效利用。噬菌体LL-H与质粒载体pX3一起可作为德氏乳杆菌乳酸亚种和保加利亚亚种菌株基因工程的有效质粒转导工具。

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