Luchansky J B, Kleeman E G, Raya R R, Klaenhammer T R
Department of Food Science, Southeast Dairy Foods Research Center, North Carolina State University, Raleigh 27695-7624.
J Dairy Sci. 1989 Jun;72(6):1408-17. doi: 10.3168/jds.S0022-0302(89)79248-1.
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.
嗜酸乳杆菌ADH是一种能产生细菌素的人体分离菌株，可黏附于人类胎儿肠道细胞和回肠细胞。我们已采用电穿孔法和接合方法将各种质粒转移至嗜酸乳杆菌ADH。此外，我们还证明了嗜酸乳杆菌ADH携带的温和噬菌体（phi adh）可在该菌株内转导质粒DNA。通过电穿孔法，质粒pGK12以高达3.3×10⁵个转化子/微克质粒DNA的频率导入ADH菌株。在与乳酸乳球菌乳酸亚种进行滤膜交配后，ADH菌株的接合子以每个供体细胞10⁻²（pAMB1）、10⁻⁴（pVA797::Tn917）和10⁻⁴（pVA797）的频率获得。质粒pGK12以平均3.4×10⁻⁸个转导子/噬菌斑形成单位的频率从噬菌体phi adh溶原菌转导至嗜酸乳杆菌ADH的受体菌株。通过表型和质粒图谱对转化子、接合子或转导子获取质粒DNA进行了验证。通过电穿孔、接合和转导转移质粒和动员DNA序列的能力将增强我们对嗜酸乳杆菌在肠道中活性进行定义和表征的努力。
