APPLICATION OF BACTERIAL ARTIFICIAL CHROMOSOMES TO THE GENERATION OF CONTIGUOUS PHYSICAL MAPS - A PILOT-STUDY OF HUMAN RYANODINE RECEPTOR GENE (RYR1) REGION

In order to increase the rate of generation of a contiguous human chro mosome 19 physical map, we have investigated the advantages of using a bacterial artificial chromosome (BAC) library in comparison with othe r systems. This cloning system, recently described, can faithfully pro pagate DNA fragments greater than 300 kb in size. We have screened a t otal human genomic BAC library with a complex cosmid probe, specific f or the ryanodine receptor gene (RYR1) located at 19q13.1. One 150-kb B AC was positive for the ryanodine receptor probe. The ryanodine recept or BAC hybridized to a 150-kb overlapping set of nine chromosome 19-sp ecific cosmids at the 3' terminus of the RYR1 gene as well as two esta blished cosmid contigs which can be linked to the same physical region . The hybridization of the BAC to a 150-kb set of chromosome 19 cosmid s suggests that this BAC is nonchimeric instructure. (C) 1994 Academic Press, Inc.