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Generating High Fidelity Sequence for the Drosophila Virilis Dot Chromosome
Author(s) -
Itano Michelle Sumako,
Kellmann Elmer,
Leung Wilson,
Slawson Elizabeth,
Shaffer Christopher,
Buhler Jeremy,
Mardis Elaine,
Elgin Sarah CR
Publication year - 2006
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.20.4.a469
Subject(s) - drosophila virilis , biology , genetics , chromosome , heterochromatin , euchromatin , drosophila melanogaster , y chromosome , microbiology and biotechnology , gene
Studies have shown that the “dot” chromosome, the small fourth chromosome of Drosophila melanogaster , is almost completely heterochromatic. Heterochromatin Protein 1 (HP1), a protein associated with heterochromatin, localizes to the dot chromosomes of Drosophila melanogaster and other closely related Drosophila species. However, D. virilis , diverged 40‐60 million years ago, has a dot chromosome that does not stain with fluorescent antibodies specific for HP1. This suggests that the D. virilis dot chromosome is euchromatic. During the past two years, portions of the D. virilis dot chromosome have been identified and finished to high quality sequence by the students in Bio 4342. At the end of the Spring 2005 semester, 960,000 bps of the D. virilis dot had been sequenced, leaving 240,000 bps to be sequenced in Spring 2006. I have worked to fill in missing regions by pulling out additional clones from the D. virilis fosmid library. This gridded library contains over 18,000 clones. PCR primers were designed from the reference D. virilis sequence (an unfinished sequence from a different strain) to create template for the creation of radioactively labeled probes that were then used against the gridded library. Fosmids that appeared positive for the probe were prepped and end‐sequenced to determine their specific locations on the D. virilis dot chromosome. The most effective series of positive fosmids was identified and prepared to be sent to the Genome Sequencing Center to be made into libraries for use in the Bio 4342 Spring 2006 lab course.