Wednesday, 20 November 2002
D0512

This presentation is part of : Display Presentations, Section B. Physiology, Biochemistry, Toxicology, and Molecular Biology

Identification of ARSs in mosquitoes functional in yeast and their use in TAR cloning

Young S. Hong1, Cheolho Sim1, Lucas Ton1, Sun-Hee Leem2, and Frank H. Collins1. (1) University of Notre Dame, Biological Sciences, Galvin Life Science, Notre Dame, IN, (2) Dong-A University, Biology, Hadan 840, Busan, South Korea

Autonomously replicating sequences (ARSs) were first found in yeast, functioning as a replication initiation site for DNA replication in eukaryotes. Despite their absence in bacteria, ARSs are structurally complex but functionally well conserved in eukaryotes so that heterologous ARSs effectively support DNA replication in yeast. We identified ARSs from four different mosquito species, Aedes aegypti, Anopheles gamgiae, Culex quinquefasciatus, and Toxorhynchites amboinensis. A sequence analysis searching for 11 bp ARS consensus (ARSC) sequence (WTTTATRTTTW) in Ae. aegypti and An. gambiae revealed the presence of ARSC. It was also observed that ARSC itself is sufficient enough to serve as an origin of DNA replication when introduced in yeast. In addition, the application of ARSs for sequence-specific isolation of large genomic DNA by homologous recombination (TAR cloning) in yeast will be discussed.

Species 1: Diptera Culicidae Anopheles gambiae
Species 2: Diptera Culicidae Aedes aegypti
Species 3: Diptera Culicidae Culex quinquefasciatus
Keywords: ARS, replication

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