Generator

Part:BBa_K173004

Designed by: Lorenzo Pasotti, Susanna Schiavi, Letizia Diamante, Paolo Magni   Group: iGEM09_UNIPV-Pavia   (2009-10-15)
Revision as of 12:17, 18 October 2009 by Lor18 (Talk | contribs)

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.

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]


[edit]
Categories
Parameters
None