Molecular cloning and functional expression of bovine brain GABA transaminase.
Jeon S.G., Bahn J.H., Jang J.S., Jang S.H., Lee B.R., Lee K.S., Park J., Kang T.C., Won M.H., Kim H.B., Kwo O.S., Cho S.W., Choi S.Y.
We isolated a cDNA that encodes the bovine brain gamma-aminobutyrate transaminase (GABA-T; EC 2.6.1.19) from the lambda gt 11 cDNA library, which showed a high degree of sequence similarity to the corresponding enzymes from various sources. Northern blot analysis revealed two differentially expressed GABA-T transcripts of approximately 2.0 and 6.0 kb in the bovine tissues. Southern blot analysis indicates that the two GABA-T transcripts are encoded in a greater-than 10-kb, single-copy gene. Bovine GABA-T cDNA was expressed in E. coli using the pGEX bacterial-expression vector system. The overexpressed GABA-T was enzymatically active after purification, and it had very similar kinetic parameters when compared with those of other mammalian GABA-Ts.
Mol. Cells 12:91-96(2001) [ PubMed | SRS | CiteXplore ]
