A method of identifying and isolating a unique member of a multigene family: application to a trypanosome surface antigen gene

Nucleic Acids Res. 1991 Apr 25;19(8):1811-5. doi: 10.1093/nar/19.8.1811.

Abstract

A chimeric oligonucleotide was constructed using DNA sequences from two distal regions of a cDNA which encodes a major surface antigen (TSA-1) of Trypanosoma cruzi. Conditions were found that allowed the chimeric oligonucleotide to hybridize only to a 5.4 kb EcoRI fragment in a Southern blot of total genomic DNA. The 5.4 kb EcoRI genomic DNA fragment has previously been shown to be located at a telomeric site, thus the studies described here directly demonstrate that the TSA-1 gene is telomeric in location. It is also shown that the chimeric oligonucleotide can be used to selectively identify recombinant lambda phage which harbor the TSA-1 gene using standard library screening procedures. Since these studies demonstrate that a chimeric oligonucleotide can be used to identify in both Southern blots and library screens a single member among the more than sixty members of the TSA-1 gene family, it seems likely that chimeric oligonucleotides may be of general use in studies involving repetitive DNA sequence families.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Base Sequence
  • Blotting, Southern
  • Chimera / genetics
  • Chromosome Mapping
  • Cloning, Molecular
  • DNA, Protozoan / isolation & purification*
  • Gene Library
  • Molecular Sequence Data
  • Multigene Family*
  • Oligonucleotide Probes
  • Plasmids
  • Trypanosoma cruzi / genetics*
  • Trypanosoma cruzi / immunology
  • Variant Surface Glycoproteins, Trypanosoma / genetics*

Substances

  • DNA, Protozoan
  • Oligonucleotide Probes
  • Variant Surface Glycoproteins, Trypanosoma