Part:BBa_K4167660
LacZ generator driven by Plac
This is a β-galactosidase protein generator with strong RBS, driven by Plac promoter. IPTG can bind to the operator site of the promoter to induce β-galactosidase expression. With the different concentration of IPTG, it can express β-galactosidase at different level. With the detection of p-Nitrophenol, the activity of β-galactosidase can be measured.
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]
β-galactosidase is encoded by LacZ gene. It is widely found in animals, plants, microorganisms and cultured cells. It can catalyze the hydrolysis of β-galactoside bonds in β-galactoside compounds and release free galactose. It can also cleave lactose to glucose and galactose.
β-galactosidase is often used as a reporter protein/enzyme for colorimetric assays. For example, β-galactosidase can cleave X-gal to yield galactose and 5-bromo-4-chloro-3-hydroxyindole. The latter is then oxidized to 5-bromo-4-chloro Indigo, a blue color product which is easily measured.
In addition, β-galactosidase can decompose p-Nitrophenyl-β-D-Galactopyranoside to produce p-Nitrophenol. The product has a characteristic absorption peak at 400 nm. The activity of β-galactosidase can be characterized by the change of absorbance value, which is used to determine the activity of β-galactosidase.
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