Modified electroporation protocol for Lactobacilli isolated from the chicken crop facilitates transformation and the use of a genetic tool.
Isolates of Lactobacillus spp. from a collection of potentially probiotic strains isolated from the crops of broiler chickens were found to be non-electrotransformable using published techniques. One strain of Lactobacillus salivarius was shown to develop electrocompetence when an overnight culture was incubated in fresh medium. The effect was enhanced if glycine was incorporated into the fresh growth medium. When these modifications were applied to a number of other crop isolates of Lactobacillus spp., electrocompetence could be detected in approximately half the strains tested. Two temperature sensitive plasmid vectors that had been used for the genetic modification of other lactic acid bacteria were introduced into a crop strain of Lb. salivarius. Both showed temperature sensitivity at 42 degrees C and above but were relatively stable at 37 degrees C. The genetic tool harbouring an IS element allowed the delivery of the plasmid to multiple independent sites in the host chromosome. Harnessing such genetic tools will facilitate the future genetic analysis of the host bacterium.
Mason
Charlene K
CK
Queen's University of Belfast, Newforge Lane, Belfast, BT9 5PX, Northern Ireland, UK.
Collins
Martin A
MA
Thompson
Keith
K
D000818:Animals; D002645:Chickens / Q000382:microbiology*; D003426:Crop, Avian / Q000382:microbiology; D003470:Culture Media / Q000737:chemistry; D004251:DNA Transposable Elements / Q000235:genetics; D004587:Electrophoresis, Agar Gel / Q000662:veterinary; D018274:Electroporation / Q000379:methods / Q000662:veterinary*; D005998:Glycine / Q000378:metabolism; D007778:Lactobacillus / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification / Q000502:physiology*; D010957:Plasmids / Q000235:genetics; D019936:Probiotics; D014170:Transformation, Genetic / Q000235:genetics*
D003470:Culture Media; D004251:DNA Transposable Elements; D005998:Glycine