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10test2answersweb - QUESTION 1a Bioinformatics ORF finder...

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QUESTION 1a Bioinformatics: ORF finder - potential reading frame? BLAST - homologues in protein data banks? Check for codon bias ? Experimental: Is this region of genome transcribed? Is there a phenotypic effect if “knock-out” of putative gene? Zoo blot (to see if homologues in other organisms)? Probe probably would cross-hybridize with simple sequences located elsewhere in genomes... & BLAST search at protein level better addresses same issue QUESTION 1b (from the classnotes) Massively-parallel pyrosequencing Cluster analysis of microarray data (transcript profiling)
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QUESTION 2 1. Strategy to generate altered protein P, followed by assay of potential interaction with protein R Affinity column chromatography is simpler approach for 2d part than ones intended for identifying an unknown “prey” protein (by screening expression libraries...) 2. Strategy to determine the precise lengths and sequences of their 5’ UTRs followed by comparative sequence analysis 5’ RACE
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