title: Ips_pini-midgut.1
status: Public on Oct 17 2003
contact.city: Reno
contact.first: Christopher
contact.middle: Ian
contact.last: Keeling
contact.email: ckeeling@mac.com
contact.state: NV
contact.address: 1664 North Virginia Street
contact.department: Department of Biochemistry/Mail Stop 330
contact.country: USA
contact.web_link: http://bioinformatics.unr.edu/beetle/
contact.institute: University of Nevada, Reno
contact.phone: 775-784-1734
organism: Ips pini
description: The Ips_pini-midgut.1 array is a cDNA-based glass microarray produced for Dr. Claus Tittiger's laboratory by the Nevada Genomics Center at the University of Nevada, Reno. The array features 768 spots representing 384 tentatively unique genes chosen from an Ips pini (the North American pine engraver beetle) EST project of anterior midgut tissue from juvenile hormone III treated (20 h) males (Eigenheer, A.L., MS thesis, University of Nevada, Reno 2002 & Eigenheer, A. L., et al., Gene 316:127-136, 2003).  Pheromone biosynthesis occurs de novo in this tissue.  cDNA was amplified using vector specific primers from selected clones.  PCR products were resuspended in Micro Spotting Solution Plus (Arrayit) and spotted onto SuperAmine microarray substrates (Arrayit) using a Virtek SDDC2 Microarrayer with a 2 x 4 array of SMP3 Stealth Micro Spotting pins (Arrayit) printing subarrays of 6 x 8 spots with 300 µm spacing.  Spotting was duplicated on each substrate with duplicate spots 9 mm apart.  DNA was bound to substrate by baking for 80 min at 80°C and then the spots were rehydrated, flash dried on a hot plate, and then crosslinked with 120 mJ of UV light (Stratagene UV Stratalinker 1800).  Clones were chosen to represent the longest clone of a cluster (Eigenheer, A.L., MS thesis, University of Nevada, Reno 2002).  After this platform was spotted, the ESTs were re-clustered with Virginia Bioinformatics Institute's ESTAP software using more stringent filtering (Eigenheer, A. L., et al., Gene 316:127-136, 2003).  Those spots without GenBank accession numbers were excluded from further clustering by ESTAP.
  Keywords: North American pine engraver beetle; anterior midgut; juvenile hormone; pheromone biosynthesis; Coleoptera; Scolytidae
  
  Publication reference:
  
  Reference Type: Book Section
  Authors: Tittiger, Claus; Keeling, Christopher I.; Blomquist, Gary J.
  Year: 2005
  Title: Some insights into the remarkable metabolism of the bark beetle midgut.
  Editor: Romeo, J.T.
  Book Title: Chemical ecology and phytochemistry of forest ecosystems
  City: Toronto
  Publisher: Elsevier
  Volume: 39
  Pages: 57-78
  Series Title: Recent Advances in Phytochemistry
  