Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Dan Ziemianowicz
Date created: 2014-10-08 11:00:00
Date modified: 2015-05-08 01:10:27
Full length beta-galactosidase (LacZ)
| Types | DnaRegion |
| Roles | Reporter engineered_region |
| Sequences | BBa_K1444017_sequence (Version 1) |
Description
Beta-galactosidase (EC 3.2.1.23) is a classic enzyme most commonly used in blue-white colony screening when used in specialized vectors. This part consists of the full length beta-galactosidase sequence which can be inserted downstream of a promoter to produce the tetrameric, 47.4kDa enzyme.Beta-galactosidase cleaves the beta-glycosidic bond of a galactose bound to an organic moiety; the physiological function is to break down beta-galactosides, such as lactose, into it's respective monnosaccharides. Beta-galactosidase can act as a reporter when provided substrates such as X-gal. The cleavage of X-gal releases galactose and the chromophoric, subtsituted indole moiety. The most common form of X-gal is 5-Bromo-3-indolyl β-D-galactopyranoside, which produces an intense blue pigment. Other versions of X-gal may be used to produce other color pigments.
Typically 40-60uL of a 20ug/mL X-gal solution, dissolved in dimethyl sulfoxide or dimethyl formamide, is spread on a plate and allowed to dry before plating the bacteria expressing beta-galactosidase. X-gal may also be added to liquid cultures or incorporated into solid agar media.