Molecular cloning and functional expression of bovine brain GABA transaminase.
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.
Jeon
S G
SG
Department of Genetic Engineering, Hallym University, Chunchon, Korea.
Bahn
J H
JH
Jang
J S
JS
Jang
S H
SH
Lee
B R
BR
Lee
K S
KS
Park
J
J
Kang
T C
TC
Won
M H
MH
Kim
H B
HB
Kwo
O S
OS
Cho
S W
SW
Choi
S Y
SY
D000612:4-Aminobutyrate Transaminase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D001921:Brain / Q000201:enzymology* / Q000502:physiology; D002417:Cattle; D003001:Cloning, Molecular; D006801:Humans; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D016415:Sequence Alignment; D014018:Tissue Distribution
D011993:Recombinant Fusion Proteins; D000612:4-Aminobutyrate Transaminase