GFP is a fluorescent protein isolated from coelenterates, such as the Pacific jellyfish, Aequoria victoria, or from the sea pansy, Renilla reniformis. The green fluorescent protein (GFP) is made up of 238 amino acids - 26.9 kDa that fluoresces green by energy transfer when exposed to blue light. In molecular biology, the GFP gene is commonly used as a reporter of expression. Basically, it has been used to make biosensors, and many animals have been created that express GFP as a proof-of-concept that a gene can be expressed throughout a given organism. The GFP gene can be introduced into organisms and maintained in their genome through breeding, or local injection with a viral vector which can be used to introduce the gene.
In molecular biology, the GFP gene is commonly used as a reporter of expression. Basically, it has been used to make biosensors , and many animals have been created that express GFP as a proof-of-concept that a gene can be expressed throughout a given organism. The GFP gene can be introduced into organisms and maintained in their genome through breeding, or local injection with a viral vector which can be used to introduce the gene.
(Ribbon diagram of the crystal structure of GDP)
How the GFP gene is incorporated into E.Coli?
( E.coli is actually a type of common bacteria..)
The GFP gene is incorporated in E.coli firstly by inserting the GFP gene into the plasmid DNA known as the pGLO. The pGLO protein also contains antibiotic resistance gene which allows only the growth of transformed bacteria in an agar containing antibiotic like ampicillin. Subsequently, transformation solution is used to heat shock the pGLO plasmid into the E.coli cell. Heat shock is often done to increase the permeability of the cell membrane and allow uptake of the pGLO plasmid into the cell. Following that, the cells are allowed to grow such that it will eventually express the GFP gene and form protein. Other features on pGLO, like most other plasmids, include: a selectable marker, Ori (origin of replication), and an MCS (multiple cloning site) located at the end of the GFP gene. The plasmid is 5371 base pairs long. In supercoiled form, it runs on an agarose gel in the 4200-4500 range.
