Difference between revisions of "Part:BBa K1444017"

Revision as of 03:32, 18 October 2014

Full length beta-galactosidase (LacZ)

Beta-galactosidase (EC 3.2.1.23), or LacZ, 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. This part was amplified from a well-used expression vector from a molecular biology lab, and cloned into the BioBrick vector pSB1C3.

Usage and Biology

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 [http://en.wikipedia.org/wiki/X-gal X-gal]. The cleavage of X-gal releases galactose and the chromophoric, substituted 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.

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]

@@ Line 2: / Line 2: @@
 <partinfo>BBa_K1444017 short</partinfo>
-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 (EC 3.2.1.23), or LacZ, 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. This part was amplified from a well-used expression vector from a molecular biology lab, and cloned into the BioBrick vector <partinfo>pSB1C3</partinfo>.
-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 &#946;-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.
 ===Usage and Biology===
-This part was amplified from a commonly used expression vector from a molecular biology lab, and cloned into the BioBrick vector <partinfo>pSB1C3</partinfo>. With expression via an appropriate promoter, this part catalyzes the cleavage of lactose into glucose and galactose. In the lab, it most commonly used to cleave the lactose analog [http://en.wikipedia.org/wiki/X-gal X-gal], which after cleavage produces a deep blue colour.
+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 [http://en.wikipedia.org/wiki/X-gal X-gal]. The cleavage of X-gal releases galactose and the chromophoric, substituted indole moiety. The most common form of X-gal is 5-Bromo-3-indolyl &#946;-D-galactopyranoside, which produces an intense blue pigment. Other versions of X-gal may be used to produce other color pigments.
-uL 20 mg/mL X-gal (2%)
+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.
-uL of 1mM IPTG dissolved in DMSO.