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1.
Article in English | IMSEAR | ID: sea-138999

ABSTRACT

Background & objectives: The SXT element, also known as ‘constin’ (conjugable, self transmissible, integrating element) is an integrating conjugative element (ICE) in Vibrio cholerae discovered in the chromosome of epidemic V. cholerae O139 strain MO10 (SXTMO10) which arose in late 1992 in Chennai, India. SXT related ICEs have become widespread and currently, most if not all Asian V. cholerae clinical isolates contain SXT related ICEs. The present study attempts to determine the presence of SXT Int gene in V. cholerae recovered between 2005 to 2007 in a tertiary care hospital, demonstrate its conjugal nature and also detect co-presence and co-transfer of plasmids in representative isolates. Methods: This prospective study was done on 116 V. cholerae isolates [114- O1 (107 ogawa and 7 inaba) and 2 - Non O1 Non O139 V. cholerae] from watery stools between 2005 to 2007 recovered from equal number of patients. PCR was carried out using SXT Int specific primers that produced a 592 bp internal fragment of SXT element, and rifampicin resistant strain of E.coli K-12 was used as recipient in conjugation experiments to study transfer of SXT, as also co-transfer of resistance to tetracycline, erythromycin, and nalidixic acid. Antibiotic susceptibility was performed against various antibiotics. Results: Of the 116 isolates, 110 (94.8%) were positive for SXT element by PCR. It was demonstrated in 94.7 per cent of the O1, and 100 per cent of non O1 non O139 V. cholerae. All 2005 isolates, 25 per cent of 2006 isolates and 96.6 per cent of 2007 isolates were positive for SXT. Thirty two drug resistance patterns were observed and the 2007 isolates showed resistance to as many as eight antibiotics. The resistance of SXT positive isolates was higher than those of SXT negative and the typical drug resistance pattern corresponding to SXTET and SXTMO10 was shown by only one V. cholerae O1 isolate. Successful conjugal transfer of SXT was seen in 31 (88.6%) of the 35 isolates studied without any co-transfer while, presence of plasmids was observed in two of the 31 donor V. cholerae studied. Interpretation & Conclusions: The demonstration of SXT element and its successful horizontal transfer in V. cholerae isolates studied emphasizes the need for its detection to monitor antibiotic resistance and dissemination in V. cholerae.


Subject(s)
Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Cholera/microbiology , DNA Transposable Elements , Humans , Interspersed Repetitive Sequences , Prospective Studies , Vibrio cholerae/drug effects , Vibrio cholerae/genetics , Vibrio cholerae/isolation & purification , Vibrio cholerae/metabolism
2.
Progress in Biochemistry and Biophysics ; (12): 986-993, 2006.
Article in Chinese | WPRIM | ID: wpr-408435

ABSTRACT

Commonly, the interbacterial transfer of circular plasmids is initiated by nicking at an internal sequence, oriT, followed by transferring one strand as single-stranded DNA through a type Ⅳ secretion channel on cell membrane. In contrast, Streptomyces conjugative linear plasmids, containing a free 3'-end but a protein-capped 5'-end, can potentially undergo cell-to-cell transfer by transfer of non-nicked DNA. It was reported that circular derivatives of the Streptomyces lividans linear plasmid SLP2, as well as the parental linear plasmid itself can transfer efficiently. And the genetic requirements for such transfer was described. Efficient transfer of plasmid requires six co-transcribed SLP2 genes, encoding a Tra-like DNA translocase, cell wall hydrolase, two cell membrane proteins that interact with an ATP binding protein, and a protein of unknown function. Reduced transfer efficiency of plasmid from SalⅠ R-/M-to Sal Ⅰ R/M hosts argues that transfer of both the circular and linear forms of the plasmid involves double-stranded DNA. These results suggest that conjugal transfer occurs by a similar mechanism for SLP2-derived linear and circular plasmids, and cellular membrane/wall functions in the transfer process.

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