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J. Biol. Chem., Vol. 260, Issue 5, 2605-2608, Mar, 1985
E Ohtsuka, S Matsuki, M Ikehara, Y Takahashi and K Matsubara
Two deoxyoligonucleotide probes (23-mer and 26-mer) carrying deoxyinosine
residues (I) at positions corresponding to ambiguous nucleotides derived
from amino acid sequence have been synthesized by the phosphotriester
method using a polymer support. The 23-mer and 26- mer corresponded to the
mRNA for 8 amino acids from gastrin and 9 amino acids from cholecystokinin,
respectively. The dIs have been used where the base in the third position
of the amino acid codon is ambiguous. These deoxyoligonucleotides were used
as probes for hybridization with colonies containing the corresponding
cDNAs or genes. The hybrid formed between a gastrin clone and the 23-mer
that harbors 5 dIs was dissociated at 50-55 degrees C, suggesting that
deoxyinosine did not significantly effect the stabilization or
destabilization of the DNA duplex. A similar result was obtained using the
26-mer that contains 5 dIs and a phage clone DNA of the cholecystokinin
gene. Thus oligonucleotide probes with deoxyinosine residues at ambiguous
points seem to be useful as hybridization probes for cloning genes for
proteins containing amino acids with degenerate codons.
An alternative approach to deoxyoligonucleotides as hybridization probes by insertion of deoxyinosine at ambiguous codon positions
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