Amplification of the phosphorylation site ‐ ATP‐binding site cDNA fragment of the Na+,K+‐ATPase and the Ca2+‐ATPase of Drosophila melanogaster by polymerase chain reaction

Abstract

In vitro DNA-amplification technique has been utilized to generate a 430 bp fragment of the Na⁺,K⁺-ATPase, and a 550 bp fragment of a Ca²⁺-ATPase (the sarcoplasmic reticulum-type) of Drosophila melanogaster. The oligonucleotide primers for the DNA-amplification (Polymerase Chain Reaction) had been designed on the basis of amino acid sequence motifs - the phosphorylation site and the ATP-binding site - conserved among members of the ATPase protein family. Using the amplified cDNA-segments as probes, we demonstrated that there is one Na⁺,K⁺-ATPase and one Ca²⁺-ATPase (sarcoplasnaic reticulum-type) gene in the Drosophila genome. Three different mRNA species are processed from the Na⁺,K⁺-ATPase gene and one from the Ca²⁺-ATPase gene. Developmental control in expression of the Ca²⁺-ATPase gene was observed.