I will map origins of DNA replication on human chromosome 22. The Snyder laboratory constructed a DNA microarray that contains approximately 85% of the nonrepetitive DNA of chromosome 22 in fragments ranging from 0.3-1.4 kilobases. I will utilize a replication timing technique to map DNA origins with the existing chromosome 22 microarray. In addition, I will construct an assymetric chromosome 22 DNA microarray by strand specific amplification of each of the DNA fragments from the microarray. Due to the bidirectional nature of DNA replication at origins, the change in polarity of leading and lagging strand synthesis indicates the presence of an intervening origin of replication. I will utilize previously published origin mapping protocols for the isolation of both leading and lagging strand (Okazaki fragment) synthesis. The isolated nacent strands will be labeled and hybridized to the assymetric array. Origins of DNA replication will be identified at sites of replication fork polarity switching. The two techniques are complementary since the replication timing technique identifies regions containing origins while fork polarity switching more accurately defines origin location.

Agency
National Institute of Health (NIH)
Institute
National Human Genome Research Institute (NHGRI)
Type
Postdoctoral Individual National Research Service Award (F32)
Project #
5F32HG002715-02
Application #
6807031
Study Section
Special Emphasis Panel (ZRG1-F08 (20))
Program Officer
Graham, Bettie
Project Start
2003-06-01
Project End
2006-05-31
Budget Start
2004-06-01
Budget End
2005-05-31
Support Year
2
Fiscal Year
2004
Total Cost
$47,296
Indirect Cost
Name
Yale University
Department
Biochemistry
Type
Schools of Arts and Sciences
DUNS #
043207562
City
New Haven
State
CT
Country
United States
Zip Code
06520
White, Eric J; Emanuelsson, Olof; Scalzo, David et al. (2004) DNA replication-timing analysis of human chromosome 22 at high resolution and different developmental states. Proc Natl Acad Sci U S A 101:17771-6