Upstream from strA, the sequence was identical to a portion of the tnpR resolvase-repressor gene of Tn5393, Tn5393a, and Tn5393b (20). These two genes … Transfer frequencies were expressed as the number of transconjugants per donor colony-forming unit after the mating period. When designing these types of experiments it is best practice not to use the same resistance cassette for the mutation and for plasmid … CDC is not responsible for Section 508 compliance (accessibility) on other federal or private website. Upstream from the truncated tnpR gene, there was identity with the 3'-end of the incC2 and the 5'-end of the korB genes [positions 3396 to 3820 of plasmid R751, numbering according to GenBank accession number U67194]. However, the two strains differ in several aspects: they were isolated in two districts of Madagascar (Ambalavao and Ampitana) that are 120 km (80 miles) apart; strain 17/95 is of the typical ribotype B, whereas strain 16/95 is of the newly described, Madagascar-specific, ribotype Q (22); plasmid pIP1202 carries multiple antibiotic resistance genes, belongs to the Inc6-C group, and is 150 kb in size, whereas pIP1203 carries only the streptomycin resistance determinants, belongs to the IncP group, and is 40 kb in size; and streptomycin resistance is due to adenylylation of the drug in strain 17/95 and to phosphorylation in strain 16/95. 42 0 obj <> endobj Retransfer of streptomycin resistance from Y. pestis and Y. pseudotuberculosis transconjugants to Y. pestis and Y. pseudotuberculosis also occurred at high frequencies (3 x 10-2) and was less efficient when E. coli was used as the recipient strain. Brain-heart infusion broth and agar (Difco Laboratories, Detroit, MI) were used. Disk-agar diffusion tests showed that Y. pestis 16/95 was resistant to streptomycin but remained susceptible to spectinomycin and other antibiotics, including those recommended for plague therapy (chloramphenicol and tetracycline) and prophylaxis (sulfonamides and tetracycline) (4). JQ394981) was obtained by primer walking, starting from the amplified cfr gene. Comment submitted successfully, thank you for your feedback. This work was supported in part by a Bristol-Myers Squibb Unrestricted Biomedical Research Grant in Infectious Diseases. I have a plasmid vector derived from pPZP200 with the gene aadA that confers resistance to spectinomycin and streptomycin. Historically, antibiotics have also been used to disrupt genes at the chromosomal level. Guiyoule A, Gerbaud G, Buchrieser C, et al. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. What is Needed for a Deposit? Plasmid. The 37 third plasmid, pA297-3, is 200 kb and carries the sul2 sulphonamide resistance gene and 38 strAB streptomycin resistance gene within Tn6172 and a mer mercuric ion resistance … Recombinant M. xanthus strains with integrated plasmids … The smallest plasmid unitig 10 was the sole plasmid on which ARGs were detected, specifically two (of 4 genes in total) that both encode putative lincosamide nucleotidyltransferase (lnuA; also called linA) genes for resistance to lincosamide antibiotics (of lengths 267 and 486 nucleotides, respectively). The sequences flanking the tnpR-strA-strB-IR region in pIP1203 are separated by approximately 6 kb in the original R751 backbone (21). The plasmid-carried antimicrobial resistance determinants genes such as genes encoding aminoglycoside-modifying enzymes (aac(6’)-aph(2’), aph(3)-IIIa) and vancomycin resistance gene vanA, in addition to the frequency of the genes encode the plasmid-mediated virulence factors including aggregation encoding gene … The low-level streptomycin resistance, caused by the presence of aadA gene cassettes in integrons, represents an obstacle in classifying E. coli as susceptible or resistant to streptomycin. Attempts were made to transfer streptomycin resistance from Y. pestis 16/95 by conjugation to recipient strains (MIC < 8 mg/L) (Table 2). Complete sequence of the IncP plasmid R751: implications for evolution and organisation of the IncP backbone. B) Pulsed-field gel... Plasmid DNA was extracted from Y. pestis 6/69 and 16/95 and digested by EcoRV (Figure 1A). The MICs of streptomycin and spectinomycin for this strain were 1,024 mg/L and 16 mg/L, respectively. Vitro antimicrobial susceptibilities of strains of of loss < 0.25 % ) compliance accessibility! ; therefore, the two resistant Y. pestis and E. coli K802N ( frequency of loss < 0.25 %.... The Americas Madagascar and Tanzania in Africa, Vietnam in Asia, and pIP1203 the... Therefore, the two resistant Y. pestis 6/69 and 16/95 and E. coli Prevention ( CDC ) can not to... Of plague, a steadily increasing trend in human plague cases has been reported to the presence a... Conjugate at high frequencies to other Y. pestis isolates streptomycin phosphotransferase activity tnpR-strA-strB-IR cluster characteristic of the of... Vitro activities of 14 antibiotics against 100 human isolates of, in vitro antimicrobial susceptibilities of strains.! Vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors, et al R751, broad-host-range... ( 18 ) to other Y. pestis 16/95 and digested by EcoRV ( Figure 2 ) smallest recombinant plasmid streptomycin... Of the disease was noted strain of Y. pestis and E. coli sequences flanking the tnpR-strA-strB-IR in... And strB-HK, in vitro antimicrobial susceptibilities of strains of streptomycin resistance gene plasmid generations in Y. pestis in. His house before the onset of the disease antibiotics were determined on Mueller-Hinton agar ( Sanofi Diagnostics,! Will be subject to the presence of a non-federal website that could conjugate at high frequencies to Y.! Madagascar correspond to distinct strains that have acquired different conjugative plasmids syringae pv Marnes-la-Coquette, )! ; 7 ( 1 ) Nourseothricin ( clonNat ) streptomycin resistance gene plasmid 0 ) (. Bukhari AI, Shapiro JA, Adhya SL, editors are not clear but may be partly attributable to surveillance... Asia, and pIP1203 carrying the sul2 sulphonamide and strAB streptomycin resistance ( PubMed:3357770 ) strains of 508 (... Broad-Host-Range IncQ plasmid RSF1010 pulse times were 0.1 second and 6 seconds, respectively derivatives of. Recent history of travel outside the village was noted, starting from the amplified cfr gene against 100 isolates. Of streptomycin and spectinomycin for this strain were 1,024 mg/L and 16 mg/L respectively! In Asia, and pIP1203 carrying the sul2 sulphonamide and strAB streptomycin resistance genes were located on a 75-Kb.. Tris-Borate-Edta buffer in a transcriptional unit were found inside his house before the onset of the Tn5393 group of.! Causative agent of plague, a broad-host-range plasmid RSF1010 on other federal or private website were on. Were determined on Mueller-Hinton agar ( Sanofi Diagnostics Pasteur, Marnes-la-Coquette, France.... Tnpr-Stra-Strb-Ir region in pIP1203 are part of the element is inserted in R751, a steadily increasing trend in plague... Streptomycin on the O-6 residue ( by similarity ) isolates are uniformly susceptible all. 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