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首页> 外文期刊>Journal of bacteriology >Characterization of the lactococcal temperate phage TP901-1 and its site-specific integration.
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Characterization of the lactococcal temperate phage TP901-1 and its site-specific integration.

机译：乳球菌温带噬菌体TP901-1的表征及其位点特异性整合。

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The temperate lactococcal phage TP901-1, induced by UV light from Lactococcus lactis subsp. cremoris 901-1, was characterized. The restriction map was found to be circular, and the packaging of TP901-1 DNA was concluded to occur by a headful mechanism. The pac region was localized on the 38.4-kb phage genome. TP901-1 belongs to the class of P335 phages (V. Braun, S. Hertwig, H. Neve, A. Geis, and M. Teuber, J. Gen. Microbiol. 135:2551-2560, 1989). Evidence is presented that the phages TP936-1 (V. Braun, S. Hertwig, H. Neve, A. Geis, and M. Teuber, J. Gen. Microbiol. 135:2551-2560, 1989) and C3-T1 (A. W. Jarvis, V. R. Parker, and M. B. Bianchin, Can. J. Microbiol. 38:398-404, 1992) are very closely related to or are identical to TP901-1. The lytically propagated TP901-1 phages were able to lysogenize both indicator strains Lactococcus cremoris 3107 and Wg2. Lysogenization resulted in site-specific integration of the phage genome into the bacterial chromosome. Only one chromosomal attB site was found in 20 independent lysogens. The attP region of TP901-1 and the attL and attR regions were cloned and sequenced. The results showed a core region of only 5 bp, in which the recombination occurs, followed after a 1-bp mismatch by a 7-bp identical region, TCAAT(T/C)AAGGTAA. This result was further verified by sequencing of the attB region obtained by PCR. An integration vector was constructed with the 6.5-kb EcoRI fragment from TP901-1 containing attP. This vector also functions in the plasmid-free strains, MG1363 and LM0230 with only one specific attB site, strongly indicating a more general use of the TP901-1-based integration vector in lactococci.
机译：温带乳球菌噬菌体TP901-1，由乳酸乳球菌亚种的紫外线诱导。 cremoris 901-1，被表征。发现限制性酶切图是圆形的，并且TP901-1 DNA的包装被认为是通过有害的机制发生的。 pac区位于38.4-kb噬菌体基因组上。 TP901-1属于P335噬菌体的类别（V.Braun，S.Hertwig，H.Neve，A.Geis和M.Teuber，J.Gen.Microbiol.135：2551-2560，1989）。证据表明噬菌体TP936-1（V. Braun，S. Hertwig，H. Neve，A.Geis，and M.Teuber，J.Gen.Microbiol.135：2551-2560，1989）和C3-T1（ AW Jarvis，VR Parker和MB Bianchin，Can.J.Microbiol.38：398-404，1992）与TP901-1密切相关或相同。裂解繁殖的TP901-1噬菌体能够裂解两种指示菌株克雷莫氏乳球菌3107和Wg2。溶原化导致噬菌体基因组到细菌染色体的位点特异性整合。在20个独立的溶原菌中仅发现一个染色体atB位点。克隆和测序TP901-1的attP区以及attL和attR区。结果显示，只有5 bp的核心区域发生重组，随后在1 bp错配后出现7 bp相同区域TCAAT（T / C）AAGGTAA。通过对通过PCR获得的attB区域进行测序，进一步证实了该结果。用含有attP的来自TP901-1的6.5kb EcoRI片段构建整合载体。该载体还可以在仅具有一个特定attB位点的无质粒菌株MG1363和LM0230中起作用，强烈表明在乳球菌中更普遍地使用基于TP901-1的整合载体。

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