(A) The fusion of nonmonomerized enhanced green fluorescent protein (EGFP) to a resident endoplasmic reticulum (ER) membrane protein induces the formation of an organized smooth ER structure. ( B ) The fusion of monomerized EGFP to the same protein does not grossly alter the structure of the ER.

In this blog I discuss strategies to generate fusion proteins and highlight some aspects of their design. The green fluorescent protein (GFP) from Aequorea victoria and its variants are genetically encoded fluorescent probes. One of the limitations is the size of GFP: ~240 amino acids or about 28 kDa.

2024年8月23日 · In order to generate a GFP fusion protein, a cDNA that encodes the protein of interest must be available or obtainable. Then, through molecular cloning, the cDNA for GFP must be cloned upstream or downstream of the cDNA for the protein of …

2017年11月24日 · Green fluorescent protein (GFP) fusions are pervasively used to study structures and processes. Specific GFP-binders are thus of great utility for detection, immobilization...

2005年12月20日 · Existing variants of green fluorescent protein (GFP) often misfold when expressed as fusions with other proteins. We have generated a robustly folded version of GFP, called 'superfolder'...

Fusion tagging: One of the most common uses, GFP can be fused to the N- or C-terminus of a protein, which allows the scientist to visualize when and where the gene is expressed. Click here to view Addgene’s collection of empty backbones for constructing fluorescent fusions.

2006年12月1日 · GFP fusion proteins can be transiently or stably expressed. Although transient expression is quick and can provide informative results, in many cases it is beneficial and/or essential to develop stable cell lines expressing the fusion protein of interest.

GFP can be detected in living cells without selection or staining and it can be fused to other proteins to yield fluorescent chimeras. The potential of GFP has also been recognised by gene therapy researchers and various GFP-tagged therapeutic proteins have been constructed.

2002年1月1日 · By allowing breathtaking visualization of fluorescent fusion proteins as they move within and between cells, GFP has fundamentally transformed the spatial analysis of protein function. Now, new...

2008年5月1日 · Here, we demonstrate a novel method, based on high specificity and affinity of monoclonal antibody against GFP, for purification of GFP and GFP fusion proteins. The binding of the monoclonal antibody to the GFP epitope allows isolation of GFP, as well as many active GFP-fused recombinant proteins, directly and easily from crude cellular sources ...