Characterization of the genes encoding integrative and excisive functions of Lactobacillus phage øg1e: cloning, sequence analysis, and expression in Escherichia coli.
øg1e is a temperate phage of the Lactobacillus strain G1e. The phage-host junctions attR and attL cloned from the lysogen have a 24-bp common (core) sequence implicated in recombination. DNA sequencing analysis of a 5.2-kbp SacI fragment of the øg1e phage genome (42.5 kbp) revealed two possible open reading frames (ORF), xis and int, and the phage attachment (recombination) site (attP), whose 24-bp sequence is identical to the core sequence detected in attR and attL. The deduced int product (Int) is a basic protein of 391 amino acids with an estimated pI of 9.70, and significantly resembles other presumed integrases encoded by the Lactobacillus and Lactococcus phages including øadh and øLC3, as well as the Escherichia coli phages such as lambda. The predicted øg1e xis protein (Xis) is small and very acidic (66 amino acids; pI 4.55), and shows a resemblance (32% overall identity) with a putative excisionase encoded by the Staphylococcus phage ø11. The øg1e Int with a deduced molecular mass of 45.5 kDa was overproduced in E. coli cells, and electrophoretically analyzed.
Kakikawa
M
M
Faculty of Engineering, Toyama University, Gofuku, Japan.
Oki
M
M
Watanabe
N
N
Yasukawa
H
H
Masamune
Y
Y
Taketo
A
A
Kodaira
K I
KI
D000595:Amino Acid Sequence; D001287:Attachment Sites, Microbiological / Q000235:genetics; D010582:Bacteriophage lambda / Q000235:genetics; D001435:Bacteriophages / Q000235:genetics*; D001483:Base Sequence; D015139:Blotting, Southern; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D004254:DNA Nucleotidyltransferases / Q000235:genetics* / Q000502:physiology; D004926:Escherichia coli / Q000235:genetics / Q000821:virology; D015964:Gene Expression Regulation, Bacterial; D019426:Integrases / Q000235:genetics* / Q000502:physiology; D007778:Lactobacillus / Q000821:virology*; D017045:Lactococcus / Q000821:virology; D008242:Lysogeny / Q000235:genetics*; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010957:Plasmids; D015183:Restriction Mapping; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013210:Staphylococcus / Q000821:virology; D014764:Viral Proteins
D014764:Viral Proteins; D004254:DNA Nucleotidyltransferases; D019426:Integrases; C063911:excisionase