Isolation and Characterization of Mouse Xrcc-1, a DNA Repair Gene Affecting Ligation
1994; Elsevier BV; Volume: 22; Issue: 1 Linguagem: Inglês
10.1006/geno.1994.1359
ISSN1089-8646
AutoresKerry W. Brookman, Robert S. Tebbs, Susan A. Allen, James D. Tucker, Roy R. Swiger, Jane E. Lamerdin, A.V. Carrano, Larry H. Thompson,
Tópico(s)RNA Interference and Gene Delivery
ResumoHuman DNA repair gene XRCC1 complements the strand-break rejoining defect in Chinese hamster mutant EM9 and encodes a protein that is apparently required for optimal activity of DNA ligase III. Toward the goal of producing transgenic mice that carry a mutation in the Xrcc-1 locus, the murine homolog of XRCC1 was cloned from both cosmid genomic and cDNA libraries. Upon transfection into EM9 cells, cosmids containing the functional mouse gene efficiently corrected (94-100%) the high sister-chromatid-exchange defect. Mouse Xrcc-1 is 26 kb in length, contains 17 exons, and maps by metaphase in situ hybridization to the 7A3-7B2 region of mouse chromosome 7. Isolated cDNA clones were highly truncated and were extended by anchored polymerase chain reactions. The 1893-bp open reading frame of mouse Xrcc-1 encodes 631 amino acids, compared with 633 for the human homolog. The predicted mouse Xrcc-1 protein of 69.1 kDa and pI of 5.95 is 86% identical and 93% similar to human XRCC1.
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