title: Purification and expression of the gene III protein from filamentous phage phi Lf.
abstract: The gene III protein (pIII) from phi Lf, a filamentous phage of Xanthomonas campestris pv.campestris, was purified by gel filtration with FPLC. The gIII coding region was amplified by PCR, which was then cloned into pUC18 and expressed in Escherichia coli. The size of both pIII, purified from phage particle and expressed in E. coli, is similar to the value deduced from the nucleotide sequence as shown by Western blot analysis. This is different from the case in Ff phages (f1, fd, and M13), in which the size of pIII observed in SDS-polyacrylamide gel electrophoresis is substantially larger than the deduced value. Upon infection of X. c. pv. vesicatoria carrying cloned phi Lf gIII with phi Xv, a filamentous phage of pv. vesicatoria, the progeny particles in supernatant were able to infect both pv. campestris carrying cloned phi Lf gIII and pv. vesicatoria, indicating that a mixture of authentic phi Xv and chimeric phage consisting of phi Xv DNA and phi Lf pIII was produced. These results suggest pIII to be the adsorption protein required for host recognition.
authors.affiliation: Institute of Molecular Biology, National Chung Hsing University, Taichung, Taiwan.
authors.forename: T J
authors.initials: TJ
authors.lastname: Liu
authors.forename: B Y
authors.initials: BY
authors.lastname: You
authors.forename: N T
authors.initials: NT
authors.lastname: Lin
authors.forename: M T
authors.initials: MT
authors.lastname: Yang
authors.forename: Y H
authors.initials: YH
authors.lastname: Tseng
mesh_terms: D001435:Bacteriophages / Q000737:chemistry* / Q000472:pathogenicity; D002213:Capsid / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*; D036022:Capsid Proteins; D004926:Escherichia coli / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis; D013045:Species Specificity; D016959:Xanthomonas campestris / Q000821:virology*
chemical_list: D036022:Capsid Proteins; D011994:Recombinant Proteins; C110214:gene III protein, bacteriophage Phi-Lf