Broad-Host Range Gene Transporter Particles Produced by Aliivibrio fischeri
Hiroshi Xavier Chiura1, Nami Uchiyama, Kazuhiro Kogure
1Marine Microbiology Laboratory, Department of Marine Ecosystems Dynamics, Ocean Research Institute, University of Tokyo.
Abstract:
Aliivibrio fischeri NCIMB1281(T) (basonym, Vibrio fischeri) spontaneously started broad-host range vector particle (AfVP) production by budding from the logarithmic phase, and stabilised at around 7.0×10(10)-7.4×10(11) particles mL(-1) without any accompanying change in the host population. AfVPs had a spherical shape and varied in diameter from 18.1 to 159.2 nm [median±SD, 58.4±11.9 nm, n=528], with 95.1% between 30.2 and 84.6 nm in diameter exhibiting a normal distribution. Their buoyant density and DNA content ranged from 1.3607 to 1.3980 g cm(-3), and 17.3 to 95.3 kbp, respectively. Regardless of UV treatment, AfVPs enhanced the efficiency of plating 116-136% at a multiplicity of infection of ca. 140 in Escherichia coli AB1157. Generalised transduction was observed with a frequency of between 10(-4) and 10(-6) cells per AfVP without UV treatment. Upon infection, the particle membrane remained outside the recipient cell, and a string-like structure coated with a fibrous proteinaceous-like material was present. The growth of the E. coli transductant (AfV-E-trans) reached a maximum of ca. 415% that of the parental E. coli recipient. AfV-E-trans acquired the ability to produce budding particles.
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