Molecular cloning and deoxyribonucleic acid polymorphisms in Lactobacillus acidophilus and Lactobacillus gasseri.
Lactobacillus strain ADH is a bile-resistant, bacteriocin-producing human isolate that was phenotypically classified within the Lactobacillus acidophilus group. Total DNA and phage DNA extracted from strain ADH were separately digested with BclI and ligated with BclI-digested pGK12. Following electroporation of these ligation mixtures directly into strain ADH, electrotransformants were recovered at frequencies of 1.5 x 10(3) and 2.0 x 10(4)/micrograms of pGK12 for preparations of pGK12::phage DNA and pGK12::total DNA, respectively. Among the electrotransformants screened, 6 and 22% contained passenger DNA of either phage DNA or chromosomal origin, respectively, as determined by restriction-enzyme analyses and hybridization assays. Derivatives of pGK12 containing passenger DNA of chromosomal (pTRK120) or phage (pTRK121) origin and pTRK15 (native cryptic plasmid) were evaluated for use as species-specific probes. The strain ADH-derived probes hybridized primarily to members of the B-1 and B-2 lactobacilli homology groups and demonstrated strain-specific polymorphisms within these groups. Identical hybridization patterns were established for strain ADH and Lactobacillus gasseri VPI 6033 (ATCC 19992). Identification of DNA probes and establishment of a host-vector cloning system have facilitated our efforts to characterize the Lactobacillus chromosome and to distinguish between closely related species thought to be important inhabitants of the gastrointestinal tract.
Luchansky
J B
JB
Southeast Dairy Foods Research Center, North Carolina State University, Raleigh 27695-7624.
Tennant
M C
MC
Klaenhammer
T R
TR
D002876:Chromosomes, Bacterial; D003001:Cloning, Molecular; D015342:DNA Probes; D004262:DNA Restriction Enzymes; D004269:DNA, Bacterial / Q000235:genetics; D016680:Genome, Bacterial; D007778:Lactobacillus / Q000145:classification / Q000235:genetics*; D007779:Lactobacillus acidophilus / Q000145:classification / Q000235:genetics*; D009693:Nucleic Acid Hybridization; D010957:Plasmids; D011110:Polymorphism, Genetic; D014170:Transformation, Genetic
D015342:DNA Probes; D004269:DNA, Bacterial; D004262:DNA Restriction Enzymes