|
KMID : 0545119960060030162
|
|
Journal of Microbiology and Biotechnology
1996 Volume.6 No. 3 p.162 ~ p.166
|
|
|
High-level Expression and Purification of Recombinant 4-Aminobutyrate Aminotransferases in Escherichia coli
|
|
Lee, Sung Gu
Choi, Tae-Jin/Kim, Young Tae
|
|
|
Abstract
|
|
|
|
The protein coding sequence of the 4-aminobutyrate aminotransferase was amplified by polymerase chain reaction (PCR) from a previously cloned cDNA of pig brain using a pair of primers based on the published sequence. The amplified DNA was introduced into a T7 expression vector. Recombinant 4-aminobutyrate aminotransferases were overexpressed in Escherichia coli. The inclusion bodies were formed when enzyme was overexpressed. The unfolded, overproduced proteins were purified by chromatography with hydroxyapatite and refolded by a sequential dialysis method. The renatured 4-aminobutyrate aminotransferase regained the catalytic activity. However, the purified mutant protein did not show the catalytic function of 4-aminobutyrate aminotransferase.
|
|
|
KEYWORD
|
|
|
|
|
|
FullTexts / Linksout information
|
|
|
|
|
|
Listed journal information
|
|
  
|