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Characterization of Filamentous Bacteriophage Lf from Xanthomonas Campestris pv. Campestris

Kuo-Chih Lin^1,

Chao-Chi Pan^1,>

¹ Department of Botany
and² Agricultural Biotechnology Laboratories, National Chung Hsing University, Taichung, Taiwan 40227, Republic of China

A filamentous phage, Lf, which specifically infects Xanthomonas campestris pv. campestris was isolated. The phage particle measured 1000 (±200) x 8 nm. It formed turbid plaques of about 1 mm in diameter. During multiplication, the progeny virions extruded into the medium without retarding host cell growth. Stocks were stable for 6 months at 4 °C and survived treatment at 80 °C for 10 min. Treatment with chloroform, ethanol or acetone completely destroyed infectivity; ethyl ether and methanol inactivated 98 to 99% of the phage. SDS-polyacrylamide gel electrophoresis showed a major coat protein band of approximate M_r 4000 whereas an immunoprecipitation test detected the existence of two coat protein species. The phage genome was shown to be a single-stranded DNA molecule. A physical map was constructed and the DNA size was calculated to be 5.9 kb. Cells treated with Tris-HCl containing CaCl₂ and polyethylene glycol 6000 were transfected by replicative form DNA at a frequency of 3.4 x 10³ p.f.u./µg.

Present address: Molecular Biology Division, Development Center for Biotechnology, Taipei, Taiwan.

^> Present address: Union Chemical Laboratories, Industrial Technology Research Institute, Hsinchu, Taiwan.

Received 3 January 1990; accepted 26 March 1990.

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