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KMID : 0357319960310050533
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Journal of the Korean Society for Microbiology
1996 Volume.31 No. 5 p.533 ~ p.546
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Construction of Ordered Overlapping Cosmid Library and Bacterial Artificial Chromosome(BAC) Library of Helicobacter pylori
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류복덕
이우곤/조명제/전영석/이광호
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Abstract
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To elucidate the host-parasite relationship on the basis of genome of Helicobacter pylori, we constructed a cosmid and a bacterial artificial chromosome(BAC) library of Helicobacter pylori isolate strain #51. Genomic DNA fragments were cloned
into
the
cosmid vector SuperCos1 and a random 868 member library, which were 15-fold redundant to Helicobacter pylori genome (~1700 kb), were constructed. The cosmids were ordered by dot blot hybridization with RNA probes specific for the ends of cloned
DNAs. Up
to now, 34 cosmids were mapped to 8 independent miniset contigs. The DNA of each cosmid in the 8 minisets was fingerprinted by restriction analysis. The sum of each length of 8 miniset contigs was about 810 kb which is considered to cover 48% of
the
Helicobacter pylori genome. A BAC library of Helicobacter pylori genome was also constructed to obtain large clones which could facilitate to assembly an ordered overlapping library. The genomic DNA of Helicobacter pylori was prepared in LMP
agarose
film, partially digested with HindⅢand then ligated with pBeloBAC11. We continue to attempt transformations of E. coli DH10B/r with these ligates and screenings of resultant colonies by in situ colony hybridization. Until now, we selected one
hundred
BAC clones which contained inserts with a range of 110~200 kb in their length.
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