TITLE	Molecular cloning and expression of the coat protein genes of Cf, a filamentous bacteriophage of Xanthomonas campestris pv. citri
AUTHOR	Mei-Kwei Yang The Graduate School of Biology, Fu-Jen University, Taipei, Taiwan, Republic of China Huei-Mei Huang Institute of Life Science, National Defense Medical Center, Taipei, Taiwan, Republic of China Yen-Chun Yang The Graduate School of Biology, Fu-Jen University, Taipei, Taiwan, Republic of China Wei-Chih Su Graduate Institute of Botany, National Taiwan University, Taipei, Taiwan, Republic of China
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ABSTRACT	Particles of the filamentous bacteriophage Cf contain a major coat protein, the B protein, with a molecular weight of approximately 6,000. In addition, a minor coat protein, the A protein, with a molecular weight of about 50,000, was also identified on sodium dodecyl sulfate-containing polyacrylamide gels. A 3.3 kbp HincII fragment derived from Cf genome was cloned into the expression plasmid pG308N, an E. coli plasmid which carries pL promoter. The recombinant plasmid pG33 and a series of deletion derivatives of pG33 were constructed and transformed into E. coli DG116 for expression of phage Cf genes. The genes coding for A and B proteins of Cf were found on the 2.0 kbp EcoRI-HincII fragment. The complete nucleotide sequences of the 2.0 kbp EcoRI-HincII insert were determined. The deduced amino acid sequence corresponds to a 62-amino acid-residue polypeptide that has a calculated Mr of 6070 was identified as the B protein by SDS/PAGE and immunoblotting. Another open reading frame (ORF419) downstream of the B protein gene (ORF62) was found, and was shown to code for a polypeptide of 419 amino acids with a calculated Mr of 44,676 that exhibits considerable identity to the A protein.
KEYWORD	Bacteriophage Cf; Coat protein genes; Gene expression; Host specificity; Nucleotide sequences;
ARTICLE INFO	Botanical Bulletin of Academia Sinica, Volume 36 Number 4 October 1995, page 207-214, 8 pages
PUBLISHER	Institute of Plant and Microbial Biology, Academia Sinica, Taipei, Taiwan, Republic of China