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嗜热链球菌温和噬菌体phi Sfi21溶原性DNA模块的特性分析。

Characterization of the lysogeny DNA module from the temperate Streptococcus thermophilus bacteriophage phi Sfi21.

作者信息

Bruttin A, Desiere F, Lucchini S, Foley S, Brüssow H

机构信息

Nestlé Research Centre, Nestec Ltd., Lausanne, Switzerland.

出版信息

Virology. 1997 Jun 23;233(1):136-48. doi: 10.1006/viro.1997.8603.

DOI:10.1006/viro.1997.8603
PMID:9201223
Abstract

Phage phi Sfi21, the only temperate Streptococcus thermophilus phage from our phage collection, showed extensive DNA homology with virulent phages from lytic group I. Southern blot hybridizations demonstrated that the phi Sfi21-specific DNA was clustered in an approximately 6.6-kb-long region, the putative lysogeny module. Sequence analysis and database research identified an integrase within this module; orf 203 with homology to an anonymous orf 258 from the temperate lactococcal phage BK5-T; orf 127 and orf 122 with weak homology to the N- and C-terminal parts, respectively, of the cl-like repressor from lactococcal phages Tuc2009 and BK5-T; orf 75 with homology to a repressor protein from lambdoid phage 434 and an anti-repressor ant with homology to phage P1. The molecular arrangement of the predicted orfs in phage phi Sfi21 was very similar to that of the lactococcal phage BK5-T. The transition from phi Sfi21-specific DNA into DNA shared with virulent phages was abrupt and flanked at one side by notable DNA repeats. Sequence analysis identified a holin protein to the left of the lysogeny module. A site-specific deletion of 2.4 kb, which reproducibly transformed phi Sfi21 into a lytic phage, was localized in the lysogeny module. It was flanked at both sides by conspicuous DNA repeats. One repeat region reflected the DNA around the attP site, while the other reflected the putative genetic switch region between repressor and anti-repressor genes. S. thermophilus host Sfi1 transformed with a plasmid containing int and orf 203 showed resistance to superinfection by heterologous phages, but not by the homologous phi Sfi21. Part of the int gene could be deleted without loss of this activity, while a deletion in orf 203 resulted in loss of the phage resistance. We speculate on the possibility of a bipartite immunity system for the control of lysogeny in phi Sfi21.

摘要

噬菌体phi Sfi21是我们噬菌体库中唯一的温和嗜热链球菌噬菌体,它与裂解I组的烈性噬菌体显示出广泛的DNA同源性。Southern杂交表明,phi Sfi21特异性DNA聚集在一个约6.6 kb长的区域,即假定的溶原模块。序列分析和数据库研究在该模块中鉴定出一种整合酶;与温和乳球菌噬菌体BK5-T的一个未知orf 258具有同源性的orf 203;分别与乳球菌噬菌体Tuc2009和BK5-T的cl样阻遏物的N端和C端具有弱同源性的orf 127和orf 122;与λ样噬菌体434的一种阻遏蛋白具有同源性的orf 75和与噬菌体P1的一种抗阻遏物具有同源性的ant。噬菌体phi Sfi21中预测的orf的分子排列与乳球菌噬菌体BK5-T非常相似。从phi Sfi21特异性DNA到与烈性噬菌体共有的DNA的转变是突然的,并且在一侧由明显的DNA重复序列侧翼。序列分析在溶原模块左侧鉴定出一种孔蛋白。一个可重复地将phi Sfi21转化为裂解性噬菌体的2.4 kb位点特异性缺失位于溶原模块中。它两侧都有明显的DNA重复序列。一个重复区域反映了attP位点周围的DNA,而另一个反映了阻遏物和抗阻遏物基因之间假定的遗传开关区域。用含有int和orf 203的质粒转化的嗜热链球菌宿主Sfi1对异源噬菌体的超感染具有抗性,但对同源噬菌体phi Sfi21没有抗性。int基因的一部分可以被删除而不丧失这种活性,而orf 203中的一个缺失导致噬菌体抗性丧失。我们推测了phi Sfi21中用于控制溶原性的二分免疫系统的可能性。

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