2000
DOI: 10.1128/jb.182.9.2445-2452.2000
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Abstract: Genetic studies in Borrelia burgdorferi have been hindered by the lack of a nonborrelial selectable marker. Currently, the only selectable marker is gyrB r , a mutated form of the chromosomal gyrB gene that encodes the B subunit of DNA gyrase and confers resistance to the antibiotic coumermycin A 1 . The utility of the coumermycin-resistant gyrB r gene for targeted gene disruption is limited by a high frequency of recombination with the endogenous gyrB gene. A kanamycin resistance gene (kan) was introduced int… Show more

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Cited by 197 publications
(220 citation statements)
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“…High-passage B. burgdorferi sensu stricto strain B31A and the derived flaB mutant MC-1 (referred to as the flaB mutant) have been described previously (9,46). Strain N40 was kindly provided by E. Fikrig, Yale University.…”
Section: Methodsmentioning
confidence: 99%
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“…High-passage B. burgdorferi sensu stricto strain B31A and the derived flaB mutant MC-1 (referred to as the flaB mutant) have been described previously (9,46). Strain N40 was kindly provided by E. Fikrig, Yale University.…”
Section: Methodsmentioning
confidence: 99%
“…Briefly, the flgE gene and the antibiotic resistance genes fused to the B. burgdorferi flgB promoter were amplified by PCR with primers P1 to P6 (Table 1), and the resultant PCR products were cloned into a pGEM-T vector (Promega). The kanamycin (kan) and streptomycin (aad) resistance cassettes were inserted into flgE at its single EcoRI site (9,18). The resultant flgE-kan and flgE-aad portions of the constructs were amplified by PCR, and approximately 5-g portions of PCR products were electroporated into competent B. burgdorferi cells.…”
Section: Methodsmentioning
confidence: 99%
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“…A single clone of avirulent B31, referred to as B31-A, was used for all gene transfer experiments and served as the reference wild-type strain (27). The nonmotile PF-deficient mutant MC-1, liquid BSK-II medium, agarose plates, growth conditions, and swarm agar plates have been described (8).…”
Section: Methodsmentioning
confidence: 99%