Molecular architecture of classical cytological landmarks: Centromeres and telomeres
Both the human telomere repeat and the pericentromeric repeat sequence (GGAAT)n were isolated based on evolutionary conservation. Their isolation was based on the premise that chromosomal features as structurally and functionally important as telomeres and centromeres should be highly conserved. Both sequences were isolated by high stringency screening of a human repetitive DNA library with rodent repetitive DNA. The pHuR library (plasmid Human Repeat) used for this project was enriched for repetitive DNA by using a modification of the standard DNA library preparation method. Usually DNA for a library is cut with restriction enzymes, packaged, infected, and the library is screened. A problem with this approach is that many tandem repeats don`t have any (or many) common restriction sites. Therefore, many of the repeat sequences will not be represented in the library because they are not restricted to a viable length for the vector used. To prepare the pHuR library, human DNA was mechanically sheared to a small size. These relatively short DNA fragments were denatured and then renatured to C{sub o}t 50. Theoretically only repetitive DNA sequences should renature under C{sub o}t 50 conditions. The single-stranded regions were digested using S1 nuclease, leaving the double-stranded, renatured repeat sequences.
- Research Organization:
- Los Alamos National Lab., NM (United States)
- Sponsoring Organization:
- USDOE, Washington, DC (United States)
- DOE Contract Number:
- W-7405-ENG-36
- OSTI ID:
- 10193973
- Report Number(s):
- LA-UR-94-3466; CONF-9403172-1; ON: DE95002710
- Resource Relation:
- Conference: US-Japan joint seminar on classical and molecular cytogenetic analysis of cereal genomes,Manhattan, KS (United States),Mar 1994; Other Information: PBD: [1994]
- Country of Publication:
- United States
- Language:
- English
Similar Records
A 37-kb fragment common to the pericentromeric region of human chromosomes 13 and 21 and to the ancestral inactive centromere of chromosome 2
Structure of DNA near long tandem arrays of alpha satellite DNA at the centromere of human chromosome 7