Identification of a replication protein and repeats essential for DNA replication of the temperate lactococcal bacteriophage TP901-1

被引:23
作者
Ostergaard, S
Brondsted, L
Vogensen, FK
机构
[1] Royal Vet & Agr Univ, Dept Dairy & Food Sci, DK-1958 Frederiksberg C, Denmark
[2] Tech Univ Denmark, Dept Microbiol, DK-2800 Lyngby, Denmark
关键词
D O I
10.1128/AEM.67.2.774-781.2001
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
DNA replication of the temperate lactococcal bacteriophage TP901-1 was shown to involve the gene product encoded by orf13 and the repeats located within the gene. Sequence analysis of 1,500 bp of the early transcribed region of the phage genome revealed a single-stranded DNA binding protein analogue (ORF12) and the putative replication protein (ORF13). The putative origin of replication was identified as series of repeats within orf13 and was shown to confer a TP901-1 resistance phenotype when present in trans. Site-specific mutations were introduced into the replication protein and into the repeats. The mutations were introduced into the TP901-1 prophage by homologous recombination by using a vector with a temperature-sensitive replicon. Subsequent analysis of induced phages showed that the protein encoded by orf13 and the repeats within orf13 were essential for phage TP901-1 amplification. In addition, analyses of internal phage DNA replication showed that the ORF13 protein and the repeats are essential for phage TP901-1 DNA replication in vivo. These results show that orf13 encodes a replication protein and that the repeats within the gene are the origin of replication.
引用
收藏
页码:774 / 781
页数:8
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