Part:BBa_K3716023
C4-LacO-gusA (E. coli)
C4-LacO-gusA (E. coli)
Usage and Biology
This biological part can express the β-glucuronidase enzyme from species Escherichia Coli. This protein can hydrolyze the glycyrrhizic acid (GL) into glycyrrhetinic acid (GA) by cutting off the glucuronide groups. This part can be used in E. coli BL21(DE3) for expression with IPTG induction.
As a composite part, C4-LacO-gusA (E. coli) contains five basic parts, of which the two most important basic parts BBa_K37160012 and BBa_K3716009 are newly built, documented, and uploaded in iGEM 2021.
BBa_K3716012 is the coding sequence of β-glucuronidase, of which all related measurement results can be referred to at at this link BBa_K3716012.
BBa_K3716009 is a C4 promote obtained from mutation of a T7 promoter. This part can be recognized by a T7 RNA polymerase and efficiently transcribe downstream genes. iBowu-China 2021 measured this part by express sfGFP or β-glucuronidase enzyme in strains such as E. coli BL21 (DE3) to produce green fluorescence, or to convert glycyrrhizic acid into glycyrrhetinic acid. In order to explore the catalytic effects of the β-glucuronidase enzyme under different protein expression levels, according to the literature [1, 2], we designed a series of promoters, C4, H9 and G6, and compare them together with a T7 promoter. They can all be activated by T7 RNA polymerase, but the strength is different from that of the T7 promoter. All measurement results related to the C4 promoter can be referred to at this link BBa_K3716009.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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