A method of identifying and isolating a unique member of a multigene family: application to a trypanosome surface antigen gene.
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.
Ruef
B J
BJ
Department of Molecular Biology and Biochemistry, University of California, Irvine 92717.
Hecht
J H
JH
Manning
J E
JE
D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D002678:Chimera / Q000235:genetics; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D016054:DNA, Protozoan / Q000302:isolation & purification*; D015723:Gene Library; D008969:Molecular Sequence Data; D005810:Multigene Family; D015345:Oligonucleotide Probes; D010957:Plasmids; D014349:Trypanosoma cruzi / Q000235:genetics* / Q000276:immunology; D014643:Variant Surface Glycoproteins, Trypanosoma / Q000235:genetics*
D016054:DNA, Protozoan; D015345:Oligonucleotide Probes; D014643:Variant Surface Glycoproteins, Trypanosoma