Clones and kit description were provided by Washington University Genome Sequencing Center. For further information see sequencing and construction of the physical map of S. typhimurium. Sequence published in: McClelland M, Sanderson KE, Spieth J, Clifton SW, Latreille P, Courtney L, Porwollik S, Ali J, Dante M, Du F, Hou S, Layman D, Leonard S, Nguyen C, Scott K, Holmes A, Grewal N, Mulvaney E, Ryan E, Sun H, Florea L, Miller W, Stoneking T, Nhan M, Waterston R, Wilson RK. Complete genome sequence of Salmonella enterica serovar Typhimurium LT2. Nature. 2001 Oct 25,413(6858):852-6. The physical map of Salmonella enterica serovar Typhimurium strain LT2 (STM) was constructed by Dan Layman, Mandeep Sekhon and the Washington University Genome Sequencing Center Fingerprinting Group by generating fingerprints from a BAC library supplied by Rod Wing and Jeff Tompkins at the Clemson University Genomics Institute (http://www.genome.clemson.edu). The library was constructed using a modified pBeloBAC11. The pBeloBAC11 vector was derived from the bacterial F factor and carries a chloramphenicol resistance marker. It is a modified version of pBAC108L with the polylinker replaced by the lacZ multiple cloning site region of pGEM3Z. The DNA of Typhimurium strain LT2 was SauIIIA digested, cloned into a BamHI site, and transformed into E. coli K12 with selection for chloramphenicol resistance (25 ug/ml). For details on the vector see Kim et. al. 1996. Genomics 34:213-214. The original library consisted of 4608 BAC clones with an average insert size of 100kb. 4285 clones were sucessfully fingerprinted using PstI, analyzed with Image (http://www.sanger.ac.uk/Software/Image) and assembled in an FPC database (http://www.sanger.ac.uk/Software/fpc). The assembled contigs were manually edited to achieve proper order and were then merged with one another to generate the largest contigs possible. To facilitate the final assembly of the physical map, a tiling path of 239 100kb in silico fragments was created(with overlap of 20 kb) from the finished STM-meld sequence and added to the FPC database. This allowed for contiguous assembly of the genome as well as verified the proper correlation between the physical and sequence maps. A minimal tiling path of BAC clones was selected from the map for end-sequencing and generation of STS markers. These clones formed the tiling path that is represented in the table below and are available from the SGSC. The clones are maintained as cells of E. coli K12 carrying the pBeloBAC11 vector with inserts of Typhimurium DNA. All clones were grown in LB + 25 ug/ml chloramphenicol and are stored at -70 degrees C in 30% glycerol. For additional information on BAC library construction and sequencing see: McClelland M, et. al. Complete genome sequence of Salmonella enterica serovar Typhimurium LT2. Nature. 2001 Oct 25,413(6858):852-6. Typhimurium pBeloBAC11 Tiling Path Clones: BAC Clones containing Typhimurium inserts from a region of the Typhimurium genome (starting and ending positions of the genome section represented are listed in the table) located by blasting BAC end-sequence against the finished STM and pSLT sequences. Source.


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