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The Detection of Transposable ampicillin-resistance in Klebsiella pneumonia | ||
| Medical Journal of Babylon | ||
| Article 1, Volume 2, Issue 4, 2005, Pages 469-476 | ||
| Author | ||
| Rabab Omran | ||
| Abstract | ||
| Genetically i stable cliiical straii of K|ebsieqa pnelnania Nal0 was isolated fion respirarofy rract infection. The isolaLe harboN large singie non-conjugative plasmid p,(1 (120kbp) which encoding multiresistance for antibiotics. Successive subcuhuring of tbe isolate for about 480 generations yielded several plasmid derivarive. All maintained ampicillin fesistance. while lhey appealed va able deleted or abolished to the othef antibiotic resistance One of these derived plasmkls (,KPr) encoding arnpicillin (Ap') and bimethoprim (Tp) .esistance was transfomred into E.coli I,/1t4294 which aheady hafborc a non-conjugatlve p1^snid pACyCISl (Cn, rd.'fhe hansfornred cells (pKP2 ah.l p,1CYC 184) were subjected to successive subculturing for about 480 genefations in ampicilljn medium at 37'C with selecrion for ampiciuin and tetracycline and chloramphenicol. Subsequently scrcening lbr plasrnid content rcvealed that the prcseoce (on agarose) ofappfoximately severr bands ofplasnid DNA rangifg beiween p,(P? d,d pACyCl3l in |i.olec|Jaf size. Each ofthese bands was eluied ffom the gel, purified aDd re-transfomed inlfo r.coli liM2 94 co]mpelent ce|.|.-The hansformed colonies resisiani io ampicillin, chloramphenicol and tetmcycline were selected. The plasmid conient of these cells revealed the presence of extra size (approximately 5kb) to the pfe transfonned DNA band @'1CyCl8'l) ^s;ndicaled by gel electrophoresis. The added DNA could represent a rmnsposable DNA fiagrnenr c,cod,q IDLi lir "e,5ta ce | ||
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