Isolation of the human insulin-like growth factor I gene using a single synthetic DNA probe

EMBO J. 1984 Feb;3(2):361-4.

Abstract

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.

MeSH terms

  • Amino Acid Sequence
  • Bacteriophage lambda / genetics
  • Base Sequence
  • DNA, Recombinant*
  • Genes
  • Genes, Synthetic
  • Humans
  • Insulin / genetics*
  • Nucleic Acid Hybridization
  • Oligodeoxyribonucleotides / chemical synthesis
  • Peptides / genetics*
  • Proinsulin / genetics
  • Protein Precursors / genetics
  • Somatomedins / genetics*

Substances

  • DNA, Recombinant
  • Insulin
  • Oligodeoxyribonucleotides
  • Peptides
  • Protein Precursors
  • Somatomedins
  • preproinsulin
  • Proinsulin

Associated data

  • GENBANK/X00362
  • GENBANK/X00462