What is the purpose of the GUS reporter gene?
The functional GUS gene produces blue coloration in plants upon integration into the plant genome. Because of the ease it provides to analyze the gene expression (as no expensive equipment is needed), GUS gene is surely plant biotechnologist’s first choice as a reporter gene.
Which is the reporter gene?
Reporter genes are those genes that when introduced into target cells (e.g., brain tissues, cancer, and circulating white cells) produce a protein receptor or enzyme that binds, transports, or traps a subsequently injected imaging probe.
Where is β glucuronidase found?
lysosome
Human β-glucuronidase is located in the lysosome. In the gut, brush border β-glucuronidase converts conjugated bilirubin to the unconjugated form for reabsorption. Beta-glucuronidase is also present in breast milk, which contributes to neonatal jaundice.
How does Gus work?
The technique utilizes the uidA gene of Escherichia coli, which codes for the enzyme, β-glucuronidase; this enzyme, when incubated with specific colorless or non-fluorescent substrates, can convert them into stable colored or fluorescent products.
What is histochemical GUS assay?
3.1 Histochemical GUS Detection Assay. In the histochemical assay, hydrolysis of X-Gluc by GUS gives an insoluble and highly colored indigo dye, visualized as a blue precipitate at the site of enzyme activity that is easily detectable ( Fig. 1 ).
What is a reporter tag?
The use of epitope tags in recombinant DNA techniques allows the detection of proteins where specific antibodies are not available. Reporter genes (also known as protein expression markers or small reporter proteins) are used in gene expression studies and provide an indication of the transcriptional activity in cells.
What is a reporter gene quizlet?
Terms in this set (35) reporter gene. – has an easily detectable expression. – not endogenously present in test system. monitor promoter activity.
What are GUS reporter lines?
The GUS reporter system (GUS: β-glucuronidase) is a reporter gene system, particularly useful in plant molecular biology and microbiology. Several kinds of GUS reporter gene assay are available, depending on the substrate used. The term GUS staining refers to the most common of these, a histochemical technique.
What is GUS expression?
β-Glucuronidase (GUS) is a very versatile reporter of gene expression that is frequently used in plant molecular biology.
How is β glucuronidase used in molecular biology?
In molecular biology, β-glucuronidase is used as a reporter gene to monitor gene expression in mammalian and plant cells. Monitoring β-glucuronidase activity through the use of a GUS assay allows determination of the spatial and temporal expression of the gene in question.
What are the symptoms of beta glucuronidase in mice?
In addition, mental retardation, short stature, coarse facial features, spinal abnormalities, and enlargement of liver and spleen are observed in surviving patients. This disease has been modeled in a strain of mice as well as a family of dogs.
Where is beta glucuronidase located in the gut?
Human β-glucuronidase is located in the lysosome. In the gut, brush border β-glucuronidase converts conjugated bilirubin to the unconjugated form for reabsorption. Beta-glucuronidase is also present in breast milk, which contributes to neonatal jaundice. The protein is encoded by the GUSB gene in humans and by the uidA gene in bacteria.
What is the activity of Tyr504 in beta glucuronidase?
The specific activity of Tyr504 in the catalytic mechanism is unclear. Through comparison to the structural data of the homologous enzyme xylanase, it has been suggested that Tyr504 of β-glucuronidase might stabilize the leaving nucleophile (Glu540) or modulate its activity.