MEETING ABSTRACTS Proceedings of the Eighth Annual Mid-Atlantic Regional Extrachromosomal Genetic Elements Conference’
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Genes encoding galactose utilization in Vibrio cholerae have been cloned into Escherichia coli host strains in order to determine their functions and organization. BarnHI-digested chromosomal DNA from 01 V. cholerae strain 26-3 was ligated into the cloning vector pACYC 184. A chimeric plasmid, pHS17, containing a 5.9-kb insert expressed galactokinase activity in a gaK mutant of E. coli. However, pHSl7 did not complement either gan or galE strains. Digestion of the 5.9-kb fragment with Hind111 and ligation into pBR322 produced plasmid pHS176, with a 2.0-kb Hind111 fragment containing the gaK determinant. On the 5.9-kb fragment the Hind111 stretch is flanked by two BamHI sites located 0.4 and 3.6 kb on each side. Since plasmid pHSl7 possesses a unique EcoRI site, another chimeric plasmid, pHS30, was constructed by inserting EcoRIdigested chromosomal DNA from V. cholerae into pBR322. The inserts of pHS17 and pHS30 overlap each other over a 4.7-kb region, which encodes the gaK gene. Plasmid pHS30 was capable of complementing a g&Kgaff strain of E. coli but not a strain with the entire gal operon deleted. Insertion of an ampicillin resistance gene into the BamHI
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تاریخ انتشار 2003