Genetic transformation in Lactobacillus sp. strain 100-33 of the capacity to colonize the nonsecreting gastric epithelium in mice.
Lactobacillus isolates able to colonize the surfaces of the nonsecreting epithelia in the stomachs of monoassociated ex-germfree mice were derived from Lactobacillus acidophilus 100-33. Strain 100-33 was originally isolated from pig feces and is unable to colonize the murine gastric epithelium. In experiments involving attempts genetically to transform the capacity to colonize the epithelium, cells of strain 100-33 were treated with muralytic enzymes and mixed with polyethylene glycol and genomic or plasmid DNA extracted from Lactobacillus fermentum RI. Strain RI was originally isolated from a conventional mouse and has the capacity to colonize the nonsecreting gastric epithelium. The mixtures containing cells, polyethylene glycol, and DNA were plated on a regeneration medium. After overnight incubation, the cells were washed from the plates and introduced by gastric gavage into germfree mice. Only mice that received regenerated 100-33 cells previously mixed with genomic DNA from strain RI had layers of gram-positive bacteria on the keratinized epithelia of their stomachs. Six isolates cultured from the washed gastric tissues of these animals were characterized. When a culture of each or a pool of cultures of the six were orally administered to germfree mice, layers of gram-positive bacterial cells were visible on the keratinized gastric epithelia of the animals within 1 to 3 weeks. Cells of all six, but not of strain 100-33, reacted with antibody made in rabbits to L. fermentum RI cells, as determined by an enzyme-linked immunosorbent assay. Nevertheless, all six had fermentation profiles identical to that of strain 100-33.(ABSTRACT TRUNCATED AT 250 WORDS)
McCarthy
D M
DM
Department of Microbiology, University of Illinois, Urbana 61801.
Lin
J H
JH
Rinckel
L A
LA
Savage
D C
DC
D000818:Animals; D000942:Antigens, Bacterial / Q000032:analysis; D004269:DNA, Bacterial / Q000235:genetics; D004352:Drug Resistance, Microbial; D004797:Enzyme-Linked Immunosorbent Assay; D004848:Epithelium / Q000382:microbiology; D005260:Female; D005285:Fermentation; D005856:Germ-Free Life; D007778:Lactobacillus / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development / Q000276:immunology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008813:Mice, Inbred ICR; D010957:Plasmids; D013270:Stomach / Q000382:microbiology*; D014169:Transformation, Bacterial
D000942:Antigens, Bacterial; D004269:DNA, Bacterial