Part:BBa_K5038018
LacZ
Usage and Biology
LacZ is the coding gene of β-galactosidase. In our design, this gene is placed downstream of LIRA system as a reporter gene. The product specifically cleaves the galactoside bond and produces a colour change of substrate.
Characterization
We added Chlorophenol red-β-D-galactopyranoside (CPRG) in the system. If β‐Galactosidase is present, it will convert the yellow-orange CPRG substrate into the chlorophenol red (CPR) , which will turn the colour of the solution from yellow to purple.
(Figure 1. Color reaction of CPRG)
Sample 1: bacterial lysate of BL21(DE3) + CPRG
Sample 2: bacterial lysate of DH5α + CPRG
Sample 3: β-gal + CPRG
LacZ constitutively expresses fully functional β-gal in E.coli BL21(DE3), but expresses defective β-gal in E.coli DH5α. As shown in Figure 1, the CPRG solution turns from yellow to purple immediately when adding bacterial lysate of BL21(DE3) and β-gal (positive control).
See BBa_K5038040for more details about LIRA_ANDgate5 with the reporter gene of lacZ.
See BBa_K5038044for more details about LIRA_H01 with the reporter gene of lacZ.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 3019
- 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 3019
- 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 3019
- 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 3019
- 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 3019
- 1000COMPATIBLE WITH RFC[1000]
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