Sequencing and Mapping of Overlapping Clones Spanning a 45 kb Region Containing the Human Lysyl Oxidase Gene
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Lysyl oxidase is a copper-dependent amine oxidase which is responsible for the crosslinking of elastin and collagen fibers in the extracellular matrix. Increased lysyl oxidase activity has been associated with many fibrotic disorders, such as atherosclerosis and cirrhosis. In addition, the lysyl oxidase gene is identical to the ras-recision gene (rrg) and possesses tumor suppressor activity. The structure of this gene IS well-characterized, but sequence data only exists for the exons and the first intron. This project used a series of previously isolated overlapping phage clones to begin sequencing of the lysyl oxidase introns and the surrounding 5' and 3' regions. Restriction fragments of 8 kb or less were created from the phage clones and ligated into plasmid vectors for sequencing. The sequence data obtained, along with restriction data, allowed for the construction of a map of a 45 kb region covering the lysyl oxidase gene as well as the 5' and 3' surrounding regions. This information is currently being used by other members of the laboratory to further understanding of the expression oflysyl oxidase in normal and disease states.