Horizontal Transfer of Chloramphenicol Resistance Plasmids from Marine associated Pseudomonas spp. to Escherichia coli JM109

M. Jayaprakashvel¹, S. Achuthan, B. Prakash², M.C. Vanitha³* and A. Jaffar Hussain³

¹Department of Marine Biotechnology, AMET University, Kanathur, Chennai - 603112, India. ²Department of Biotechnology, PGP Arts and Science, Namakkal - 637207, India. ³Centre for Marine Bioprospecting, AMET University, Kanathur, Chennai - 603112, India.

DOI : http://dx.doi.org/10.13005/bbra/1385

ABSTRACT: The present study has concentrated on the antibiotic resistance pattern and transferability of the plasmid mediated resistance of marine bacteria. A total of 9 different samples comprising of rhizosphere of coastal sand dune plants and sea water were used in the study to isolate fluorescent pseudomonads (FPs). From these samples pseudomonas were isolated by spread plate technique using Kings B agar about 60 FPs were isolated and were screened primarily for pigment production in KB agar. When screened for antibiotic resistance the isolates AMET6109 and AMET6152 have exhibited average growth in the presence of high concentration of chloramphenicol .For plasmid transfer experiments a total of nine donor strains (Pseudomonas sp) were tested for three different donor recipient (Escherichia coli JM109) ratio. The co cultured bacteria were spread plated on EMB agar amended with Chloramphenicol (100 μg/mL concentration). To demonstrate the transformed chloramphenicol resistant plasmid from E.coli, the transformed E.coli was streaked on antibiotic amended EMB agar plate. The ability of transformed bacterium in subsequent generations in the antibiotic amended medium denotes the survival of the transferred plasmid.

KEYWORDS: Antibiotic resistance; Pseudomonas spp.; curing; Escherichia coli JM109

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