Sequencing and DNA Fingerprint Mapping of Overlapping Clones Spanning a 45kb Genomic Region Containing the Human Lysyl Oxidase Gene
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|Krawetz, Stephen A.
|v, 31 p.
|Lysyl oxidase is an extracellular, copper dependent enzyme that initiates the covalent cross-linking of collagen and elastin. These cross-linkages, which provide tensile strength and elasticity, are essential to the normal function and structural integrity of the connective tissue matrix. Increased lysyl oxidase activity has been reported in fibrotic diseases including hypertension, atherosclerosis, and hepatic and pulmonary fibrosis, while deficiencies oflysyl oxidase have been observed in cutis laxis, and X-linked recessively inherited disorders such as Menkes syndrome and Ehlers-Danlos syndrome subtype IX. Another interesting facet of lysyl oxidase is that it is identical to the ras-recision gene (rrg) and possesses tumor suppressor activity. Sequence data exists only for the exons and the first introns, while the structure of the gene has been well characterized. This project began to uncover the sequence data 5' and 3' to the gene along with that of the introns to lay the foundation for further studies of the regulatory sites and controls of the gene. Overlapping restriction fragments made from previously obtained YAC clones were sequenced using both primers and nested deletion techniques. A fingerprint digest was also performed, and together with the sequence data and restriction data, allowed for the improvement of a previously made map covering the region 5' and 3' to the gene as well as the 45kb region covering the human lysyl oxidase gene. The information obtained by this project is currently being used by the lab to sequence the whole region in hopes that this information will be used in the near future as a form of genetic therapy for fibrotic diseases and cancers.
|Department of Obstetrics/Gynecology. Wayne State University. Detroit, Michigan.
|Kalamazoo College Biology Senior Individualized Projects Collection
|Senior Individualized Projects. Biology;
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|Sequencing and DNA Fingerprint Mapping of Overlapping Clones Spanning a 45kb Genomic Region Containing the Human Lysyl Oxidase Gene