Strategy for selection of internal, low redundancy probes for identification of specific cDNA.
Internal peptide fragments containing tryptophan residues are useful predictors of coding sequence for selection of restriction enzyme fragments or synthetic oligonucleotides to use in isolation of a cDNA or genomic clone. We describe a strategy to identify such fragments which uses an on-line photodiode array spectrophotometric analysis of tryptic fragment elution from an HPLC system to select peptides for amino acid sequence analysis. We applied this information to the identification and subsequent isolation of a cDNA corresponding to bovine placental lactogen from a bovine placental cDNA library which contains numerous closely related gene products.
Department of Dairy Science, University of Wisconsin, Madison 53706.
K
K
Shimomura
L A
LA
Schuler
R D
RD
Bremel
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002417:Cattle; D004247:DNA / Q000032:analysis*; D015342:DNA Probes; D006868:Hydrolysis; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D009841:Oligonucleotides / Q000032:analysis; D010446:Peptide Fragments / Q000032:analysis; D010449:Peptide Mapping; D010928:Placental Lactogen / Q000235:genetics*; D011388:Prolactin / Q000235:genetics; D012689:Sequence Homology, Nucleic Acid; D013053:Spectrophotometry; D014357:Trypsin; D014364:Tryptophan / Q000032:analysis
D000596:Amino Acids; D015342:DNA Probes; D009841:Oligonucleotides; D010446:Peptide Fragments; D014364:Tryptophan; D011388:Prolactin; D004247:DNA; D010928:Placental Lactogen; D014357:Trypsin