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SUMMARY:Microarrays and molecular biology: beyond gene expression profilin
 g - Dr. Paul Bertone\, EBI
DTSTART:20061115T140000Z
DTEND:20061115T150000Z
UID:TALK5864@talks.cam.ac.uk
CONTACT:Danielle Stretch
DESCRIPTION:This talk will describe the development and application of two
  novel microarray technologies\, focusing on genomic DNA tiling arrays and
  protein-based arrays. Tiling arrays are designed to represent all of the 
 DNA comprising a chromosome or other genomic locus\, without prior consult
 ation of existing gene annotation. Because they are developed for the unbi
 ased interrogation of genomic sequence\, tiling arrays enable the discover
 y of novel transcripts and regulatory elements in eukaryotic genomes. Two 
 examples of microarray-based genome annotation are presented: large-scale 
 mapping of transcribed sequences and identification of transcription facto
 r-binding sites. Chromosome- and genome-wide transcriptional activity is a
 ssessed by probing tiling arrays with normal complex tissue RNA\; the loca
 tions of cis-regulatory elements are determined via array hybridisation to
  chromatin-immunoprecipitated DNA\, originally bound in vivo by a transcri
 ption factor of interest. These approaches are first demonstrated with amp
 licon arrays representing all of the non-repetitive DNA of human chromosom
 e 22\, then extended to the entire genome using maskless photolithographic
  DNA synthesis technology. A large-scale tiling array survey revealed the 
 presence of thousands of novel transcribed sequences in addition to known 
 and predicted genes\, providing the first global transcription map of the 
 human genome. The second part of the talk will discuss the development of 
 protein-based microarrays for the large-scale characterisation of biochemi
 cal activity. Full-length\, functional proteins are produced from a librar
 y of expression clones\, purified as GST::HisX6::fusions and immobilised t
 o various support surfaces at high spatial density. This technology enable
 s high-throughput screening for protein interactions with various analytes
  and chemical libraries\, using procedures analogous to those developed fo
 r DNA microarrays. The method was first explored in a pilot study to assay
  the protein kinases of the budding yeast Saccharomyces cerevisiae using a
  silicone elastomer microwell array prototype\, then extended to the entir
 e yeast proteome using conventional contact-printing microarray technology
 . This platform is used to assess protein-protein\, protein-nucleic acid a
 nd small-molecule interactions\, enzymatic activity and posttranslational 
 modifications on a proteome-wide scale.\n
LOCATION:MR5\, DAMTP
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