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Allows for quantitative measurement of GUS enzyme activity in transformed plants through use of a fluorogenic substrate.
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The β-glucuronidase (GUS) enzyme from E. coli (EC 18.104.22.168) has been well documented to provide desirable characteristics as a marker gene in transformed plants. The GUS reporter gene system has many advantages including stable expression of E. coli GUS enzyme, no interference with normal plant metabolism, and low intrinsic GUS activity in higher plants. The enzyme is also capable of tolerating amino-terminal additions, making it useful for study of plant organelle transport [1,2,3,4]. Various β-glucuronic acid substrates are available for detection of GUS expression, all of which contain the sugar D-glucopyranosiduronic acid attached by glycosidic linkage to a hydroxyl group of a chromogenic, fluorogenic, or other detectable molecule . The most widely used fluorogenic substrate for detection of β-glucuronidase activity in vitro is 4-methylumbelliferyl β-D-glucuronide (4-MUG). Upon hydrolysis by GUS, the fluorochrome 4-methylumbelliferone (7-hydroxy-4-methyl coumarin) is produced along with sugar glucuronic acid . It is used in this fluorescent activity detection test. Please contact our technical services department for further information.
- Jefferson RA, Burgess SM, Hirsh D. (1986) “beta-Glucuronidase from Escherichia coli as a gene fusion marker.” Proc Natl Acad Sci USA. 83: 8447-8451.
- Jefferson RA, Kavanagh TA, Bevan MW. (1987) “GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants.” EMBO J 6: 3901-3907.
- Kosugi S, Ohashi Y, Nakajima K, Arai Y. (1990) “An improved assay for beta-glucuronidase in transformed cells: methanol almost completely suppresses a putative endogenous for beta-glucuronidase activity.” Plant Sci 70: 130-140.
- Gallagher SR (1992) GUS Protocols: Using the GUS Gene as a Reporter of Gene Expression. Academic Press, Inc
- Horvath BM, Magyar Z, Zhang Y, Hamburger A, Bako L, Visser RGF, Bachem CWB, Bogre L. (2006) “EBP1 regulates organ size through cell growth and proliferation in plants.” EMBO J 25: 4909-4920