What is GFP encode?

The GFP cDNA consists of 730 bp, which encode a 238 amino acid protein with a molecular weight of 27 kD (2). Wild-type GFP emits a vibrant green fluorescence upon exposure to blue light (450-490 nm). The signal is detectable by fluorescence microscopy and fluorescence-activated cell sorting (FACS) (3).

What is GFP plasmid?

The pGLO plasmid is an engineered plasmid used in biotechnology as a vector for creating genetically modified organisms. The plasmid contains several reporter genes, most notably the green fluorescent protein (GFP) and the ampicillin resistance gene. GFP was isolated from the jelly fish Aequorea victoria.

What is the purpose of the GFP gene in the plasmid?

Green fluorescent protein (GFP) is a protein that glows with a bright green fluorescence under ultraviolet light. First isolated from the marine jellyfish Aequorea victoria, the gene encoding GFP is used in cellular and molecular biology as a reporter to detect gene expression in transgenic organisms.

How is GFP inserted into a plasmid?

To prepare the plasmid, you will receive an already existing ring of DNA, the expression plasmid, which we have cut open for you. The GFP gene is ligated into the open plasmid to recreate a circle. To ligate, an enzyme called T4 ligase is used. The outcome is a closed plasmid containing our GFP gene.

How is GFP detected?

Flow cytometry and fluorescent microscopy are two conventional tools to detect the GFP signal; flow cytometry is an effective and sensitive technique to quantitatively analyze fluorescent intensity, while fluorescent microscopy can visualize the subcellular location and expression of GFP.

What is the purpose of arabinose?

Arabinose acts as an allosteric regulator of AraC, changing which DNA sites it binds to and how it forms a dimer. Remember that arabinose is the sugar that gets catabolized by the proteins of the AraBAD operon. When arabinose is added to the environment in which E. coli live, it binds tightly to AraC.

Is GFP made of DNA?

Gfp refers to the gene that produces green fluorescent protein. Using DNA recombinant technology, scientists combine the Gfp gene to a another gene that produces a protein that they want to study, and then they insert the complex into a cell.

What is the function of EGFP?

Split EGFP: An alternative to FRET, split EGFP has also been used to study protein-protein interactions. In this case, two portions of EGFP are fused to the proteins of interest, and when they come into close proximity, the two halves of EGFP undergo folding, maturation, and fluorescence.

How is GFP used in plasmid DNA transfection?

GFP encoding plasmid DNA (25 µg) Developed and manufactured by Altogen Biosystems Kit includes 25 µg of GFP plasmid DNA DNA transfection control for fluorescent microscopy Transfection control for chemical transfection and electroporation Validated for use in human cell lines Can be used for plasmid DNA / GFP siRNA co-transfection

How is green fluorescent protein ( GFP ) used in biology?

This application note describes how the green fluorescent protein (GFP) expressed from Bio-Rad’s pGLO plasmid can be used to help illustrate and teach the central dogma of biology, from the transformation of DNA to the expression of a protein to the visualization of a trait.

Which is prokaryotic organism expresses the GFP protein?

In the first part of the exercise, a plasmid encoding the GFP protein is transformed into E. coli, a common prokaryotic organism used for DNA propagation and protein expression. Colonies of E. coliare qualitatively examined for fluorescence, which suggests that the pGLO gene is being expressed.

Where can I find the GFP nucleotide sequence?

This is the sequences that i usually use for my cloning and i also use the Snap gene software for visualize the map…