CONSTRUCTION AND CHARACTERIZATION OF A CO SMID LIBRARY OF THE CORYNEBACTERIUM-GLUTAMICUM ATCC13032 GENES

A representative genomic library of the Corynebacterium glutamicum ATC C 13032 genes in a cosmid vector Lorist6 was created. The cosmids cont ain inserts of bacterial DNA obtained by partial digestion with the Sa u3A I restrictase. Five hundred and thirty individual primary recombin ant clones were transferred into the wells of microtiter plates, where they are now being preserved. The average size of the bacterial DNA i nserts determined via a sum of restriction fragment sizes of recomibin ant molecules is about 38 kb. The capacity of the obtained gene librar y is 8.4 equivalents of the C. glutamicum genome, i. e., every fragmen t of the genome is on average represented by eight clones and is prese nted in at least one clone with the probability >99%. Clone grids (set s of recombinant clones located on the hybridization membrane in regul ar and reproducible order) were created. Specificity of the created cl one library and its representativeness were confirmed experimentally b y hybridization of clone grids with DNA probes corresponding to unique regions of the Corynebacterium genome. A plasmid containing the pheA prephenate dehydratase gene, olygonucleotide corresponding to the lysC gene, and the 21 RNA probe obtained from the insert ends in different cosmids were used as probes. The created set of clones allows the con struction of a cosmid contig overlapping the C. glutamicum genome and a physical genetic map on its base.