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For the purpose of protein interaction mapping, predicted protein-encoding ORFs were amplified by PCR precisely between the predicted (WS9 version of WormPep) initiation and termination codons, using a cDNA library (AD-wrmcDNA library - Walhout etal. Methods Enzymol. 2000;328:575-92) as template. The resulting 11,984 Gateway cloned ORFs along with the attempted ones were transfered into a two-hybrid Destination vector downstream of the vector sequence encoding the activation domain (AD) of the yeast GAL4 transcription factor. Those constructs were pooled together to constitute a 'normalized' AD-ORFeome1.1 library. Reference - Reboul J, Vaglio P etal C. elegans ORFeome version 1.1: experimental verification of the genome annotation and resource for proteome-scale protein expression. Nat Genet. 2003 May;34(1):35-41. PMID: 12679813)
Nucleotide
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