title: Development of 112 unique expressed sequence tags from chicken liver using an arbitrarily primed reverse transcriptase-polymerase chain reaction and single strand conformation gel purification method.
abstract: In order to provide information on chicken genome expression, expressed sequence tags (ESTs) were developed from chicken liver RNAs using a method based on arbitrarily primed reverse transcription-polymerase chain reaction (RT-PCR) of total RNAs. The method is similar to differential display, using one base anchored oligo-d(T) reverse-primers and 20-mer arbitrary forward-primers. A purification step by single strand conformation gel electrophoresis was added before sequencing. With a ratio of 112 unique sequences out of 155, we found this method to be highly effective when compared with EST production with randomly selected clones from non-subtracted, non-normalized libraries. A large proportion of the ESTs sequenced correspond to genes involved in transcriptional and post-transcriptional events. Cytogenetic mapping was performed for a subset of ESTs and four regions of conserved synteny between chicken and human were confirmed.
authors.affiliation: Unité Mixte de Recherche INRA-ENSA de Génétique Animale, Rennes Cedex, France.
authors.forename: W
authors.initials: W
authors.lastname: Carré
authors.forename: C
authors.initials: C
authors.lastname: Diot
authors.forename: V
authors.initials: V
authors.lastname: Fillon
authors.forename: R P
authors.initials: RP
authors.lastname: Crooijmans
authors.forename: S
authors.initials: S
authors.lastname: Lagarrigue
authors.forename: M
authors.initials: M
authors.lastname: Morrisson
authors.forename: A
authors.initials: A
authors.lastname: Vignal
authors.forename: M A
authors.initials: MA
authors.lastname: Groenen
authors.forename: M
authors.initials: M
authors.lastname: Douaire
mesh_terms: D000818:Animals; D002645:Chickens / Q000235:genetics*; D020224:Expressed Sequence Tags; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008099:Liver / Q000378:metabolism; D012313:RNA / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D026801:Synteny
chemical_list: D012313:RNA