What technique is used for the purification of green fluorescent protein?

Chromatography
Chromatography is a powerful method used in the biotechnology industry for separating and purifying proteins of interest from bacterial proteins.

How do you make green fluorescent protein?

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.

How do you test for green fluorescent proteins?

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.

Why is it important to purify GFP?

GFP has often been used as a marker of gene expression, protein localization in living and fixed tissues as well as for protein targeting in intact cells and organisms. Our data suggests this method is more efficient than any currently available method for purification of GFP protein.

How is green fluorescent protein purification by organic extraction?

An alternative method is described for the GFP purification without affinity extensions using organic extraction yielding a highly homogeneous protein indistinguishable in spectroscopic properties from that purified by previous methods. Photochem. Photobiol. Rev., 4 ( 1979), pp. 1 – 57

Where can I find recombinant green fluorescent protein?

The recombinant GFP was transiently expressed in an active form in agoinoculated Nicotiana benthamiana leaves by using Tobacco mosaic virus (TMV) RNA-based overexpression vector (TRBO). The yield of recombinant GFP was up to ~ 60% of total soluble proteins (TSP).

How long does it take to purify GFP protein?

The purification process takes only ~ 4 h and can recover 34.1% of the protein. The purity of purified GFP was more than 95% and there were no changes in its spectroscopic characteristics.

How is the purification of recombinant GFP done?

In order to achieve a high level of purity, diverse chromatographic techniques have been used to purify the recombinant GFP. In general, these chromatographic methods involve multistep, time-consuming and complicated operations, resulting in a higher purification cost [ 5 ].