syringae pv. Among the Sm Sa-resistant strains, the frequency of nonconjugative r (Sm Sa) plasmids was much higher than that of conjugative R plasmids encoding double resistance. No adenylyltransferase activity was found (data not shown). h�bbd``b`Z$��F ��H� �l ��HHOs��v�30R��ϸ�@� ��
The arrow indicates the extra large-size DNA fragment in strain 16/95. A portion of this plasmid was derived from a plasmid made by The streptomycin/spectinomycin resistance cassette is derived from pBSL15-Omega (Lindemann, … Plasmid maps are often not helpful: I have seen the aadA gene in pENTR223.1 annotated with SmR (Streptomycin Resistance, like in SnapGene) or SpnR (Spectinomycin Resistance, like in the maps from the ORFeome collection). Hemanta Koley To clone the streptomycin-resistant determinant, DNA from plasmids pIP1203 and pUC18 was partially digested with Sau3AI and BamHI, respectively, ligated, and introduced into E. coli JM109 (MIC of streptomycin = 2 mg/L). Analysis of plasmid pIP1203. All material on this website is protected by copyright, Copyright © 1994-2021 by WebMD LLC. The pCK41 plasmid consists of potential virulence genes, transposases, plasmid maintenance genes, antibiotic-resistance genes (including kanamycin, tetracycline, and streptomycin), conjugal transfer genes, and unknown ORFs. endstream
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Extracted plasmids were electrophoresed for 2 h at 35 mA on a 0.85% agarose gel in TBE buffer (89 mM Tris-base - 89 mM boric acid - 2.5 mM disodium RDTA) as described by Sambrook et al. Antimicrobial resistance (AMR or AR) occurs when microbes evolve mechanisms that protect them from the effects of antimicrobials. IR, inverted repeat; tnpA, transposase; res, resolution site; tnpR, resolvase; IS1133 and IS6100, insertion sequences; korB and incC, genes homologous to those involved in regulation and partition of plasmid R751, respectively; oriV, origin of vegetative replication of R751. To comment please, Comments on Medscape are moderated and should be professional in tone and on topic. Figure 1. Two potentially coding sequences of 801 bp and 834 bp identified in the insert were identical to the strA and strB genes that encode an aminoglycoside 3"-O-phosphotransferase [APH(3")-Ib] and a 6-O-phosphotransferase [APH[6]-Id], respectively[17]. We report the emergence of the plasmid-mediated oxazolidinone resistance gene poxtA in E. faecium from different farms in China. Plasmid DNA was extracted from Y. pestis 6/69 and 16/95 and digested by EcoRV (Figure 1). to high-level streptomycin resistance in lactic streptococci resides on the chromosome, and its expression is affected by a plasmid. A. Guiyoule is a research technician. Please confirm that you would like to log out of Medscape. The restriction fragments in strain 6/69 correspond to those of the three Y. pestis resident plasmids, pPla, pYV, and pFra[12]. Schematic representation of the regions of Tn5393 and derivatives and of plasmids RSF1010, and pIP1203 carrying the strA and strB genes. You've successfully added to your alerts. Comparison of the restriction profiles of strains 6/69 and 16/95 revealed that the latter contained at least one extra large-size EcoRV fragment corresponding to an additional plasmid, designated pIP1203. I have a plasmid vector derived from pPZP200 with the gene aadA that confers resistance to spectinomycin and streptomycin. 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]. These results suggest that another mechanism of streptomycin resistance was work-ing in addition to the direct usage of streptomycin. The streptomycin resistance genes in pIP1203 are part of the tnpR-strA-strB-IR cluster characteristic of the Tn5393 group of transposons. Resistant microbes are more difficult to treat, requiring higher doses, or alternative medications which may prove more toxic. From this streptomycin-sensitive plasmid vector recombinant molecules can thus be directly selected. An 81-bp sequence identical to the inverted terminal repeat (IR) of Tn5393 was identified downstream from pIP1203 strA-strB genes. Several plasmids from Staph. There are three primarily Amygdaloideae-infecting clades of E. amylovora.. rpsL-based streptomycin resistance was found most commonly in the Western North American clade of E. amylovora.. The IncP-1beta plasmid pB8, which confers resistance to amoxicillin, spectinomycin, streptomycin, and sulfonamides, was previously isolated from a sewage treatment plant. Direction of gene transcription is indicated by arrows. h��X�r�F���C?�TJs[�J�J-'�숊����HD @c�e����M��~�b5�r��r��ۑ�8�� ���yIȢ����"������X��3�X�2?�R&8|�0� r1�0�|&B�)��&^�u�p���FM9��8�˺/�\�G+�!ή����{1 Attempts were made to transfer streptomycin resistance from Y. pestis 16/95 by conjugation to recipient strains (MIC ≤ 8 mg/L) (Table 2). Upstream from strA, the sequence was identical to a portion of the tnpR resolvase-repressor gene of Tn5393, Tn5393a, and Tn5393b[20]. [ … You will receive email when new content is published. pA297-3 transferred sulphonamide, streptomycin and mercuric ion resistance at high frequency to a susceptible A. baumannii recipient, and contains several genes potentially involved in conjugative … The smallest recombinant plasmid conferring streptomycin resistance, pAT709, contained an 11-kb insert. 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]. The latter plasmids included also the class 1 integron In369, comprising dfrA1b and aadA1b cassettes, and a streptomycin resistance module, consisting of sul2, strA, and strB. The tnpR-strA-strB-IR region of pIP1203, which displayed a Tn5393-like organization, was flanked on both sides by sequences highly similar to portions of the broad-host-range IncP plasmid R751[21]. The subclone DNA hybridized with enzyme-digested genomic or plasmid DNA of three of 17 other resistant strains of X. c. vesicatoria as well as with the DNA from streptomycin-resistant strains Psp 34 and Psp 36 of Pseudomonas syringae pv. No incompatibility with prototype plasmids of incompatibility groups Inc FI, FII, I1, I2, N, 6-C, 7-M, 10-B-O, J, T, and W was observed[15]. Viva la (bacterial) resistance! Plasmid-mediated high-level resistance to multiple antibiotics was reported in a clinical isolate of Yersinia pestis in Madagascar in 1997. papulans, and Xanthomonas campestris pv. This recombinant plasmid conferred high levels of resistance to the new host (MIC of streptomycin = 512 mg/L) by synthesis of a streptomycin phosphotransferase. Plasmids associated with resistance to streptomycin, to streptomycin plus chloramphenicol or to tetracycline in Staphylococcus hyicus isolated from the skin of pigs have been compared, by restriction endonuclease digest patterns, with similar staphylococci from human sources and with published DNA base sequences. This website also contains material copyrighted by 3rd parties. If you log out, you will be required to enter your username and password the next time you visit. In experiments performed by reciprocal conjugation to assess the incompatibility group, pIP1203 exhibited strong incompatibility with plasmid RP4 (data not shown), which belongs to the IncP group. 0
vesicatoria[19,20]. High-level resistance was due to the presence of a streptomycin phosphotransferase activity. The size of the molecular weight markers in kilobases is indicated at the left of the gels. B) Pulsed-field gel electrophoresis of pIP1203 DNA extracted from Escherichia coli K802N transconjugant and digested by EcoRV (1), EcoRI+EcoRV (2), and EcoRI (3). The streptomycin resistance module has originated from plasmid RSF1010 (Yau et al., 2010), as indicated by IncQ-derived segment (nts 23,451–27,940 in pMCR_170_D1) containing repAC operon found next to it. h�b```f``����cB�F������;(*01�v```lvM"�0a��� A`Y930�� �ف�l�F>�椫��&��'�X\����fqu��0 O� K����T�Y� �t#A
"Improved Vector, pHSG 664, for direct Streptomycin-Resistance Selection: cDNA cloning with G:C-Tailing Procedure and Subcloning of Doubledigest DNA fragments." syringae. Streptomycin (1) Nourseothricin ... Other (0) 33 Results 0 — 25 of 33. TheSacl site is located 138 bp downstream of the translational … papulans, Pantoea agglomerans and a yellow‐pigmented, non‐fluorescent Pseudomonas sp. Address for correspondence: Elisabeth Carniel, Laboratoire des Yersinia, Institut Pasteur, 28 rue du Dr. Roux, 75724 Paris Cedex 15, France; fax: 33-1-40-61-30-01; e-mail: carniel 2@pasteur.fr. This IR is always present at the same position in the various genetic structures that carry the str genes (Figure 2). Nucleotide sequence analysis of plasmid pO26-CRL Complete DNA sequence analysis of pO26-CRL revealed the plasmid is 111,481 bp in size (Fig. 2). %%EOF
The streptomycin-resistance locus was found to be on a plasmid of 68 kb. They have been subsequently found as part of transposon Tn5393 and related elements in phytopathogenic Erwinia amylovora, Pseudomonas syringae pv. 2001;7(1) © 2001 Centers for Disease Control and Prevention (CDC). About one third of streptomycin resistance in Escherichia coli is mediated by APH-(3″). The approximate molecular mass of each plasmid was determined … These results suggest that pCK41 is a virulence plasmid of substantial importance in the E. tarda pathogenesis to fish. The plasmid has a copy number of about 25 per cell, and International Journal of Antimicrobial Agents, 2010. Plasmid pMG1encodes resistancetogentamicin, streptomycin, sulfonamides, andmercuricionsandalsomobilizespRO161,atransfer-deficientplasmidderived fromRP1. The term antibiotic resistance (AR or ABR) is a subset of AMR, as it applies to bacteria that become resistant to antibiotics. Ischemic Stroke May Hint at Underlying Cancer, Topol: US Betrays Healthcare Workers in Coronavirus Disaster, The 6 Dietary Tips Patients Need to Hear From Their Clinicians. Sequence determination of the insert in pAT710 revealed that the base composition of this fragment was 57.5% G+C, much higher than the average G+C content of the Y. pestis genome (46%)[16]. A) Agarose-gel electrophoresis of EcoRV-digested plasmid DNA from representative Yersinia pestis strain 6/69 (1) and from streptomycin-resistant strain 16/95 (2). 51 0 obj
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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. Plasmid-mediated streptomycin and sulfamethoxazole resistance in Shigella flexneri 3a. Inventors: TOBA ... Amsterdam T. HASHIMOTO-GOTOH et al. Hence you need to know exactly what spectinomycin resistance gene your plasmid carries if you want to replace spectinomycin with the cheaper streptomycin. streptomycin resistance of the pathogens have been affect-ed by frequent usage of streptomycin (Table 1). Seven novel plasmids were identified and confirmed ranging in size from 2.9 to 34.7 kbp. %PDF-1.6
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Low-level resistance was usually indicative of small multicopy RC plasmids, such as pS194, which carries the streptomycin adenyltransferase-encoding gene str. These factors cannot get inserted into the bacterial chromosome. This portion of the element is inserted in R751, a broad-host-range plasmid belonging to the IncP group. Please use this form to submit your questions or comments on how to make this article more useful to clinicians. Two streptomycin-resistant strains of B. subtilis harbored plasmids of 5.2 X 10 (6) and 9 X 10 (6), respectively, which were, however, not correlated with the antibiotic resistance. Plasmid pIP1203 was stable after 100 generations in Y. pestis 16/95 and E. coli K802N (frequency of loss <0.25%). endstream
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The third plasmid, pA297-3, is 200 kb and carries the sul2 sulphonamide resistance gene and strAB streptomycin resistance gene within Tn 6172 and a mer mercuric ion resistance module elsewhere. However, 44.1% of the isolates from non-sprayed orchards were also resistant to streptomycin. Streptomycin resistance in strains of Pseudomonas syringae pv. Genetic organization of the strA-strB genes. Like Tn5393a, pIP1203 possessed the TAG motif, which represents a putative insertion target for IS1133[20]. pages 125-128 * Totality * Attorney, Agent or Firm: Klein, Otto (DE) … To clone the streptomycin-resistant determinant, DNA from plasmids pIP1203 and pUC18 was partially digested with Sau 3AI and Bam HI, respectively, ligated, and introduced into E. … Annie Guiyoule, Guy Gerbaud, Carmen Buchrieser, Marc Galimand, Institut Pasteur, Paris, France; Lila Rahalison, Suzanne Chanteau, Institut Pasteur, Antananarivo, Madagascar; Patrice Courvalin, Elisabeth Carniel, Institut Pasteur, Paris, France. Adding an antibiotic resistance gene to the plasmid solves both problems at once – it allows a scientist to easily detect plasmid-containing bacteria when the cells are grown on selective media, and provides those bacteria with a pressure to keep your plasmid.
pS194 is a naturally occurring Staphylococcus aureus plasmid encoding streptomycin resistance. Ophthalmologic Manifestations of Escherichia Coli, 'Breakthrough Finding' Reveals Why Certain COVID Patients Die, Understanding Messenger RNA and Other SARS-CoV-2 Vaccines, COVID-19 Vaccine Has Potential Side Effects, Nurse Volunteer Says, After COVID Infection, Antibodies Highly Protective for Months, Prospective Study Shows, CDC Shortens COVID-19 Quarantine Time, With Conditions. PBP1, which in addition mediates sulfonamide resistance, has been found to be disseminated in … We describe a second Y. pestis strain with high-level resistance to streptomycin, isolated from a human case of bubonic Please enter a Recipient Address and/or check the Send me a copy checkbox. The str genes were originally described in the small, nonconjugative, broad-host-range IncQ plasmid RSF1010[18]. Cloningandcharacterization ofthe streptomycin resistance genes frompPSR1.The 1.5-kb SacI-EcoRVfragment, which contains the Smr genes strA and strB from the broad-host-range plasmid RSF1010 (43), was used to identify homolo-gous sequences on plasmids from Cus Smr and Cur Smr strains ofP. (11). Click the topic below to receive emails when new articles are available. Plasmid isolation Streptomycin-resistant transconjugants were screened for the presence of plasmid by the method of LeBlanc and Lee (10). Meaning of Resistance Plasmid: Resistance factors or resistance plasmids are a group of conjugative plasmids which confer to their bacterial host resistance to specific antibiotics, such as, chloramphenicol, sulphonamide, streptomycin, tetracycline etc., and to several metal ions, e.g., Hg, As, Cd etc. To submit your questions or comments on how to make this article more useful clinicians... 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Was identified downstream from pIP1203 strA-strB genes Amsterdam T. HASHIMOTO-GOTOH et al will be required to enter your username password. Lactic streptococci resides on the chromosome, and its expression is affected by a plasmid pRO161 acquires streptomycin genes... Broad-Host-Range plasmid belonging to the direct usage of streptomycin … streptomycin resistance was usually indicative small. Adenyltransferase-Encoding gene str to high-level streptomycin resistance of the element is inserted in R751, a broad-host-range belonging! Low-Level resistance was due to the inverted terminal repeat ( IR ) of Tn5393 and and! Dna fragment in strain 16/95 papulans, Pantoea agglomerans and a yellow‐pigmented, non‐fluorescent Pseudomonas sp comments responses! Of transposons check the Send me a copy checkbox upon mobilization, pRO161 acquires streptomycin resistance in strains of syringae... 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This IR is always present at the left of the isolates from non-sprayed orchards were resistant... A copy checkbox originally described in the various genetic structures that carry the str were... Insertion target for IS1133 [ 20 ] be required to enter your username and password the time. Send me a copy checkbox Control and Prevention ( CDC ) in the,. Insert into pUC18, which generated pAT710 a putative insertion target for IS1133 [ 20 ] Pantoea agglomerans a. A broad-host-range plasmid belonging to the presence of plasmid by the plasmid in... Resides on the chromosome, and pIP1203 carrying the strA and strB genes have a plasmid of importance. Form to submit your questions or comments on Medscape are moderated and should professional... More toxic recombinant molecules can thus be directly selected what antibiotic concentration of streptomycin resistance was due the...