Problems associated with the use of highly degenerated oligonucleotide probes. Identification of mRNA and cDNA clones corresponding to the gene for the alpha subunit of Na+,K+-ATPase

Oligonucleotides deduced from the amino acid sequence of a hexapeptide Lys-Asp-Phe-Ala-Glu-Asn were synthesized and used as probes to screen a pig kidney cDNA library for a specific DNA sequence coding for the alpha-subunit of Na+, K+-ATPase. It was shown that the mixed oligoprobe, consisting of 64...

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Published inBioorganicheskaia khimiia Vol. 11; no. 12; p. 1636
Main Authors Petrukhin, K E, Grishin, A V, Arsenian, S G, Broude, N E, Grinkevich, V A
Format Journal Article
LanguageRussian
Published Russia (Federation) 01.12.1985
Subjects
Amino Acid Sequence
Animals
Base Sequence
Chromatography, High Pressure Liquid
Cloning, Molecular
DNA - genetics
Genes
Kidney Medulla - enzymology
Nucleic Acid Hybridization
Oligonucleotides - genetics
Peptides - genetics
RNA, Messenger - genetics
Sodium-Potassium-Exchanging ATPase - analysis
Sodium-Potassium-Exchanging ATPase - genetics
Swine
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Summary:Oligonucleotides deduced from the amino acid sequence of a hexapeptide Lys-Asp-Phe-Ala-Glu-Asn were synthesized and used as probes to screen a pig kidney cDNA library for a specific DNA sequence coding for the alpha-subunit of Na+, K+-ATPase. It was shown that the mixed oligoprobe, consisting of 64 heptadecamers, could be only suitable for mRNA blot analysis. To identify the clones with specific cDNA inserts, mixed oligoprobes were fractionated by HPLC technique. For the same purpose a new set of oligonucleotides, synthesized as four groups of 16 different heptadecamers each, was used.
ISSN:0132-3423