Part:BBa_K4628011
GUS
The gus gene is present in the genomes of some bacteria, such as E. coli. It encodes β-glucosidase, a hydrolyzing enzyme that uses β-glucosinolates as substrates and whose product is readily detectable by the naked eye. Since the vast majority of plants do not have detectable background activity for glucosinolase, this gene is a commonly used reporter in plants.
Usage and Biology
The histochemical assay for GUS uses 5-bromo-4-chloro-3-indole-β-glucosiduronic acid (X-Gluc) as the reaction substrate, and the material being examined is soaked in a buffer containing the substrate. If the tissue cell undergoes transformation of the gus gene and expresses Gus, the enzyme hydrolyzes X-Gluc to produce a blue product under suitable conditions. This is an indigo dye formed by the oxidative dimerization of the initial product, which gives a blue color to the site or locus of Gus activity, which can be seen with the naked eye or under a microscope, and to a certain extent reflects Gus activity depending on the depth of staining.
Expression of GUS:
We stained tobacco tissues exogenously expressing the GUS gene by immersing them in x-Gluc staining solution for more than 1 hour. Then the chlorophyll of the plant tissue was removed by immersion in 95% ethanol. Finally, the expression of GUS gene was directly observed by naked eye.
Fig 1. Expression of GUS in Nicotiana benthamiana.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
biology | Escherichia coli |
chassis | Nicotiana benthamiana |