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Detection of human leukocyte interferon alpha a and interferon alpha 2 genes in genomic dnas by the use of deoxyoctadecyloligonucleotide probes


, : Detection of human leukocyte interferon alpha a and interferon alpha 2 genes in genomic dnas by the use of deoxyoctadecyloligonucleotide probes. Journal of Interferon Research 8(1): 51-60

Two deoxyoctadecyloligonucleotides complementary to the sequence spanning a single base substitution between human leukocyte interferon (HuIFN) .alpha.A and .alpha.2 genes were efficiently used as probes to distinguish between HuIFN-.alpha.A and -.alpha.2 genes. At 37.degree. C or 42.degree. C under aqueous conditions (0.9 M NaCl), hybridization between both probes and the .alpha.A and .alpha.2 genes without any mismatch was strong, whereas the hybridization with one base mismatch (.alpha.A probe-.alpha.2 gene and .alpha.2 probe-.alpha.A gene) was very weak of negligible. Because the single base substitution of G in the .alpha.2 gene for A in the .alpha.A gene provides an extra HinfI site in the .alpha.2 gene at the center of the sequence hybridizing to the .alpha.2 probe, digestion with HinfI restriction endonuclease caused complete loss of the hybridization between the .alpha.2 probe and the .alpha.2 gene. PvuII digestion provides 298-bp fragments hybridizing to the probes only from the .alpha.A and .alpha.2 genes among the known HuIFN-.alpha. genes. Thus, with the use of these oligonucleotide probes in combination with PvuII and PvuII-HinfI restriction endonuclease digestion, the existence of the sequences corresponding to both IFN-.alpha.A and IFN-.alpha.2 genes in human genomic DNAs was demonstrated. The results also surprisingly indicate that these genes, formerly considered alleles because of their essential identity (1 base pair difference in the coding sequencing), are not likely to be alleles, but represent closely related distinct genes.

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