Difference between revisions of "Part:BBa K173004"

Revision as of 12:19, 18 October 2009

Beta-galactosidase protein generator

Beta-galactosidase protein generator with strong RBS.

This part takes PoPS as input to express lacZ gene (BBa_I732005), encoding for beta-galactosidase enzyme. This enzyme can be used to cleave lactose molecule to glucose and galactose (see Fig.1), but can also be used as a reporter protein for colorimetric assays (together with X-Gal or ONPG as a substrate).

X-gal is cleaved by β-galactosidase yielding galactose and 5-bromo-4-chloro-3-hydroxyindole. The latter is then oxidized into 5,5'-dibromo-4,4'-dichloro-indigo, an insoluble blue product (see Fig.2 and Fig.3).

Fig.1: lactose cleavage to glucose and galactose.

Fig.2: X-Gal cleavage to galactose and an insoluble blue product.

Fig.3: example of blue colonies bearing lacZ.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

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 This part takes PoPS as input to express lacZ gene (<partinfo>BBa_I732005</partinfo>), encoding for beta-galactosidase enzyme. This enzyme can be used to cleave lactose molecule to glucose and galactose (see Fig.1), but can also be used as a reporter protein for colorimetric assays (together with X-Gal or ONPG as a substrate).
-X-gal is cleaved by β-galactosidase yielding galactose and 5-bromo-4-chloro-3-hydroxyindole. The latter is then oxidized into 5,5'-dibromo-4,4'-dichloro-indigo, an insoluble blue product.
+X-gal is cleaved by β-galactosidase yielding galactose and 5-bromo-4-chloro-3-hydroxyindole. The latter is then oxidized into 5,5'-dibromo-4,4'-dichloro-indigo, an insoluble blue product (see Fig.2 and Fig.3).
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