The genes of the 70kD and 33kD subunits of the bovine brain vacuolar proton pump have been successfully expressed in E. Coli. The fragment of the gene of 70kD subunit was obtained by polymerase chain reaction (PCR) from an isolated cDNA encoding the 70kD subunit of the bovine brain vacuolar proton pump. The fragment of the gene of 33kD subunit was also obtained by PCR from the bovine brain cDNA library. Two clones encoding 70kD and 33kD subunits were individually obtained by directly joining the PCR products into PET vector. Sodium dodecyl sulfate (SDS)-poly-acrylamide gel electrophoresis (PAGE)and Western blot analysis of cultured transformants demonstrated high expression.