Nucleotide sequence determination, characterization and purification of the single-stranded DNA-binding protein and major coat protein of filamentous phage phi Lf of Xanthomonas campestris pv. campestris.
phi Lf is a filamentous bacteriophage of Xanthomonas campestris pv. campestris, which can integrate its genome into the host chromosome. The nucleotide sequence of an EcoRV-SphI fragment (1018 bp) from the phi Lf replicative form DNA was determined. Four contiguous open reading frames (ORFs), orf98-orf43-orf38-orf42, were revealed. ORFs 98 and 42 were identified as the genes encoding a single-stranded DNA-binding protein (Sbp) and a major coat protein, respectively. Sbp was purified and found to bind with a high affinity to ssDNA prepared from phi Lf phage particles. The major coat protein showed sequence features similar to those of the typical major coat proteins of other filamentous phages. However, it appears to be synthesized as a mature product, similar to the situation with Pf3 but different from that with other filamentous phage major coat proteins which are synthesized as pre-coats and are subject to post-translational processing. The M(r)s, estimated by SDS-PAGE, of both the Sbp (10.9K) and the major coat protein (4.1K) coincide with the values deduced from the nucleotide sequences. ORFs 43 and 38 are proposed to be the genes encoding two minor coat proteins. The order of these four genes is similar to that found in the Escherichia coli filamentous phages.
Wen
F S
FS
Department of Botany, National Chung Hsing University, Taiwan, Republic of China.
Tseng
Y H
YH
D000595:Amino Acid Sequence; D001435:Bacteriophages / Q000737:chemistry / Q000235:genetics*; D001483:Base Sequence; D002213:Capsid / Q000235:genetics* / Q000302:isolation & purification; D003062:Codon; D004277:DNA, Single-Stranded / Q000235:genetics* / Q000302:isolation & purification; D004279:DNA, Viral / Q000235:genetics* / Q000302:isolation & purification; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000302:isolation & purification; D004720:Endonucleases; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016366:Open Reading Frames / Q000235:genetics; D016959:Xanthomonas campestris
D003062:Codon; D004277:DNA, Single-Stranded; D004279:DNA, Viral; D004268:DNA-Binding Proteins; D004720:Endonucleases