[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 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.
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics*; D005796:Genes; D007679:Kidney Medulla / Q000201:enzymology; D009693:Nucleic Acid Hybridization; D009841:Oligonucleotides / Q000235:genetics*; D010455:Peptides / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics*; D000254:Sodium-Potassium-Exchanging ATPase / Q000032:analysis / Q000235:genetics*; D013552:Swine
D009841:Oligonucleotides; D010455:Peptides; D012333:RNA, Messenger; D004247:DNA; D000254:Sodium-Potassium-Exchanging ATPase
Problemy, sviazannye s ispol'zovaniem oligonukleotidnykh zondov s bol'shoĭ stepen'iu vyrozhdennosti. Identifikatsiia mRNK i klonov kDNK, sootvetstvuiushchikh genu alpha-sub''edinitsy Na+,K+-ATP-azy.