Amplification of the phosphorylation site - ATP-binding site cDNA fragment of the Na +,K +-ATPase and the Ca 2+-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 Ca 2+-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 Ca 2+-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 Ca 2+-ATPase gene. Developmental control in expression of the Ca 2+-ATPase gene was observed.