Ordered Cloned DNA Map of the Genome of Vibrio cholerae 569B and Localization of Genetic Markers

Abstract

By using a low-resolution macrorestriction map as the foundation (R. Majumder et al., J. Bacteriol. 176:1105–1112, 1996), an ordered cloned DNA map of the 3.2-Mb chromosome of the hypertoxinogenic strain 569B of Vibrio cholerae has been constructed. A cosmid library the size of about 4,000 clones containing more than 120 Mb ofV. cholerae genomic DNA (40-genome equivalent) was generated. By combining landmark analysis and chromosome walking, the cosmid clones were assembled into 13 contigs covering about 90% of theV. cholerae genome. A total of 92 cosmid clones were assigned to the genome and to regions defined by NotI,SfiI, and CeuI macrorestriction maps. Twenty-seven cloned genes, 9 rrn operons, and 10 copies of a repetitive DNA sequence (IS1004) have been positioned on the ordered cloned DNA map.

Retracted:wbeT sequence typing and IS1004 profiling ofVibrio choleraeisolates

“…In our study, a 3AE5 kb band was uniformly detected in all strains (100%) which might be attributed to the incomplete copy of IS1004 in the rfb locus. It has been indicated that the copy numbers of IS1004 element in V. cholerae O1 El Tor strains are different from classical strain 569B, containing 5-6 and 10 copies, respectively (Bik et al 1996;Chatterjee et al 1998). IS1004 profiles of all V. cholerae O1 strains in our study (El Tor biotype) were identical and indistinguishable from V. cholerae O1 ATCC14035 (classical biotype).…”

“…IS1004 is an active mobile genetic element and is able to transpose in V. cholerae O1 classical and El Tor strains which has been exclusively described as an epidemiological marker (Chatterjee et al 1998;Nesper et al 2000). It comprises of about 628 bp, with no terminal inverted repeats (Bik et al 1996) which has been applied successfully for strain differentiation of V. cholerae strains.…”

Comparison of global transcription responses allows identification of Vibrio cholerae genes differentially expressed following infection

“…al. [11] in our laboratory. The control and experimental hybridizations were performed at least thrice by reusing the same blot.…”

“…Each cosmid clone contained an insert of V. cholerae genomic DNA between 20 and 40 kb. The restriction map of each clone was veri-¢ed with the original data [11], to check for deletions. The clones containing rrn operons were excluded as they gave saturated values and caused errors during calculations.…”

“…Ampicillin was used at 100 Wg ml 31 for E. coli and 50 Wg ml 31 for V. cholerae. The 92 sets of cosmid clones in Lorist M vector [11] were grown in TB medium containing phosphate bu¡er. Ampicillin (100 Wg ml 31 ) and kanamycin (30 Wg ml 31 ) were used where appropriate.…”