INVESTIGADORES
RAYA Raul Ricardo
artículos
Título:
Genetic transfer systems for delivery of plasmid deoxyribonucleic acid to Lactobacillus acidophilus ADH: conjugation, electroporation, and transduction.
Autor/es:
J. B. LUCHANSKY, E. G. KLEEMAN, R. R. RAYA, AND T. R. KLAENHAMMER
Revista:
JOURNAL OF DAIRY SCIENCE
Editorial:
AMER DAIRY SCIENCE ASSOC-ADSA
Referencias:
Lugar: Champaign, IL, USA; Año: 1989 vol. 72 p. 1408 - 1417
ISSN:
0022-0302
Resumen:
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.