EC Number   |
|---|
    3.1.2.4 | CHY1 cDNA cloned behind the 35S promoter in a binary vector, introduced into homozygous chy1-10 mutant plants by Agrobacterium tumefaciens strain GV3101 transformation |
| 3.1.2.4 | DNA and amino acid sequence determination and analysis, expression in Escherichia coli BL21(DE3) as His-tagged enzyme |
| 3.1.2.4 | expression in Escherichia coli |
| 3.1.2.4 | open-reading frame of the Chy1 gene amplified, cDNA ligated into the vector pEX5-CT/TOPO TA to create a fusion of the open-reading frame with a His tag-coding extension at the C-terminus. Plasmid transferred into Escherichia coli BL21 (DE3) cells for expression |
| 3.1.2.4 | subcloning in Escherichia coli strain XL1-Blue, and expression in Escherichia coli strain BL21 (DE3) under control of the T7 promoter, co-expression with the beta-ketothiolase gene from Ralstonia eutropha strain H16 and the (S)-3-hydroxybutyryl-CoA dehydrogenase gene from Ralstonia eutropha strain H16, or Clostridium acetobutylicum strain ATCC824 leads to biosynthesis of enantiomerically pure (S)-3-hydroxybutyric acid in the engineered Escherichia coli cells, extracellular product, method optimization, overview |
