Bacteriophage P4282, a parasite of Ralstonia solanacearum, encodes a bacteriolytic protein important for lytic infection of its host

Citation
H. Ozawa et al., Bacteriophage P4282, a parasite of Ralstonia solanacearum, encodes a bacteriolytic protein important for lytic infection of its host, MOL GENET G, 265(1), 2001, pp. 95-101
Citations number
23
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Molecular Biology & Genetics
Journal title
MOLECULAR GENETICS AND GENOMICS
ISSN journal
1617-4615 → ACNP
Volume
265
Issue
1
Year of publication
2001
Pages
95 - 101
Database
ISI
SICI code
1617-4615(200103)265:1<95:BPAPOR>2.0.ZU;2-C
Abstract
To enhance bacterial wilt resistance in tobacco expressing a foreign protei n, we isolated the bacteriolytic gene from a bacteriophage that infects Ral stonia solanacearum The bacteriolytic protein of phage P4282 isolated in To chigi Prefecture was purified from a lysate of R. solanacearum M4S cells in fected with the phage, and its bacteriolytic activity was assayed by follow ing the decrease in the turbidity of suspensions of R. solanacearum M4S cel ls. The molecular weight of the bacteriolytic protein was approximately 71 kDa, and the sequence of the N-terminal 13 amino acids was determined. We u sed oligonucleotide probes based on this amino acid sequence to isolate the bacteriolytic gene from phage P4282 DNA. This gene of 2061 bp encodes a pr oduct of 687 amino acids, whose calaculated molecular weight was 70.12 kDa. The bacteriolytic gene was placed under the control of an inducible promot er, and the plasmid was transformed into Escherichia coli NM522. The solubl e proteins extracted from E. coli NM522 cells harboring the plasmid with th e bacteriolytic gene showed obvious bacteriolytic activities against severa l strains of R. solanacearum isolated in various districts in Japan. DNA fr agments from five phages, isolated in Niigata. Aomori, Okinawa, Fukushima a nd Yamaguchi Prefectures, hybridized to the bacteriolytic gene of phage P42 82. These observations indicate that the bacteriolytic protein shows nonspe cific activity against R. solanacearum strains, and a sequence similar to t hat of the bacteriolytic gene is conserved in the DNA of other bacteriophag es. These results indicate that the generation of transgenic (tobacco) plan ts expressing the bacteriolytic gene of phage P4282 might result in enhance d resistance to bacterial wilt in tobacco.