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Genetic Transfer Systems for Delivery of Plasmid Deoxyribonucleic Acid to Lactobacillus acidophilus ADH: Conjugation, Electroporation, and Transduction¹

Departments of Food Science and Microbiology, Southeast Dairy Foods Research Center, North Carolina State University, Raleigh 27695-7624

⁴ Corresponding author.

ABSTRACT

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 (øadh) harbored by L. acidophilus ADH. Plasmid pGK12 was introduced into strain ADH by electroporation at frequencies as high as 3.3 x 10⁵ transformants/µg 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-matings with Lactococcus lactis ssp. lactis. Plasmid pGK12 was transduced from a phage øadh 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.

¹ Paper Number 11824 of the Journal Series of the North Carolina Agricultural Research Service, Raleigh 27695-7601. The use of trade names in this publication does not imply endorsement by the North Carolina Agricultural Research Service nor criticism of similar ones not mentioned.

² Department of Food Science.

³ Department of Microbiology.