Isolation of the human insulin-like growth factor I gene using a single synthetic DNA probe.
A single synthetic oligonucleotide was employed as hybridization probe to detect and enable isolation of the human insulin-like growth factor I (IGF-I) gene from a human genomic DNA library. The synthetic oligonucleotide probe coded for the B-chain of IGF-I and was designed for expression in Escherichia coli. Despite numerous interspersed mismatches, the synthetic probe hybridized specifically with seven recombinant lambda phage containing almost the entire B-chain region of the human IGF-I gene. The usefulness of this approach was further demonstrated by the detection of lambda phage containing human preproinsulin, using A and B chain synthetic oligonucleotides, 90 and 63 nucleotides in length, as hybridization probes. The nucleotide sequence of the human IGF-I exon suggests that IGF-I is synthesized as a larger precursor molecule.
D000595:Amino Acid Sequence; D010582:Bacteriophage lambda / Q000235:genetics; D001483:Base Sequence; D004274:DNA, Recombinant; D005796:Genes; D005813:Genes, Synthetic; D006801:Humans; D007328:Insulin / Q000235:genetics*; D009693:Nucleic Acid Hybridization; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis; D010455:Peptides / Q000235:genetics*; D011384:Proinsulin / Q000235:genetics; D011498:Protein Precursors / Q000235:genetics; D013002:Somatomedins / Q000235:genetics*
D004274:DNA, Recombinant; D007328:Insulin; D009838:Oligodeoxyribonucleotides; D010455:Peptides; D011498:Protein Precursors; D013002:Somatomedins; C013737:preproinsulin; D011384:Proinsulin