Genetic transfer systems for delivery of plasmid deoxyribonucleic acid to Lactobacillus acidophilus ADH: conjugation, electroporation, and transduction.
Lactobacillus acidophilus ADH is a bacteriocin-producing human isolate that adheres to human fetal intestinal cells and human ileal cells. We have employed both electroporation and conjugation methodologies to transfer various plasmids to L. acidophilus ADH. Furthermore, we have demonstrated transduction of plasmid DNA within this strain by a temperate bacteriophage (phi adh) harbored by L. acidophilus ADH. Plasmid pGK12 was introduced into strain ADH by electroporation at frequencies as high as 3.3 X 10(5) transformants/micrograms of plasmid DNA. Transconjugants of strain ADH were recovered at frequencies of 10(-2) (pAMB1), 10(-4) (pVA797::Tn917), and 10(-4) (pVA797) per donor cell after filter-mating with Lactococcus lactis ssp. lactis. Plasmid pGK12 was transduced from a phage phi adh lysogen into a recipient strain of L. acidophilus ADH at an average frequency of 3.4 X 10(-8) transductants/pfu. Transformants, transconjugants, or transductants were verified by both phenotype and plasmid profile for acquisition of plasmid DNA. The ability to transfer plasmids and mobilize DNA sequences by electroporation, conjugation, and transduction will augment our efforts to define and characterize the activities of L. acidophilus in the intestinal tract.
Luchansky
J B
JB
Department of Food Science, Southeast Dairy Foods Research Center, North Carolina State University, Raleigh 27695-7624.
Kleeman
E G
EG
Raya
R R
RR
Klaenhammer
T R
TR
D003227:Conjugation, Genetic; D004269:DNA, Bacterial / Q000235:genetics* / Q000378:metabolism; D007779:Lactobacillus acidophilus / Q000235:genetics*; D010957:Plasmids; D014161:Transduction, Genetic; D014162:Transfection
D004269:DNA, Bacterial