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Thursday, February 22, 2024

Understanding the Basics of eGFP mRNA

Vernal offers a catalog of eGFP mRNA optimized for high expression of the enhanced green fluorescent protein (eGFP). Isolated initially from jellyfish Aequorea victoria, eGFP is a widely used reporter molecule for monitoring protein localization.

What is the eGFP mRNA Sequence?

eGFP is a codon-optimized mRNA sequence that encodes the enhanced green fluorescent protein (EGFP) from the jellyfish Aequorea victoria. EGFP is an incredibly bright green fluorescent protein with a maximal emission peak at 509 nm when excited with UV light. This makes it an ideal candidate for direct detection, as it is easily visible in mammalian cells under UV light.

EGFP has a traditional b-barrel structure, and secondary structure analysis using DSSP indicates that 47% of the protein is helical. Specifically, the core helix containing the chromophore has been modeled as a mixture of the 310 and conformations.

eGFP mRNA sequence is an essential tool in many experiments, and it is commonly used as a marker gene to identify cells expressing a particular construct. 

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What is the eGFP Protein Sequence?

The eGFP gene encodes for an enhanced green fluorescent protein (EGFP), which binds to a GTP-cyclic AMP complex and emits a bright green fluorescence when illuminated with blue or ultraviolet light. This protein is commonly used as a reporter gene in cellular and molecular biology studies, allowing researchers to visualize the activity of various genes or proteins within a cell directly.

EGFP is one of the most famous reporters in life sciences research and has been utilized to monitor the localization of fusion proteins in live cells, enabling researchers to observe the function of specific promoters or pathways. For example, a cellular assay using EGFP and the autophagy-related protein BafA1 has been used to evaluate the ability of this protein to halt autophagosomal fusion with the lysosome.

While many newer and more advanced fluorescent protein variants have been developed, EGFP remains the workhorse for most applications due to its broad availability and relatively low cost. This may be partially attributed to the fact that these newer fluorescent proteins require extensive purification to obtain high levels of stability and brightness. This process is only sometimes feasible in many laboratories.

A common concern with these newer fluorescent proteins is the propensity of specific variants to oligomerize and produce an excess of dim light. This is a problem because faint light results in lower luminosity, which can limit the amount of signal detected by microscopy. However, researchers have been able to mitigate this issue by incorporating mutations that reduce the occurrence of oligomerization in their fusion protein designs.

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What is the eGFP Molecular Weight?

Green Fluorescent Protein (GFP) is a widely used marker for protein localization. It was initially isolated from the jellyfish Aequorea victoria and is now one of the most popular tools for direct detection in mammalian cells. GFP can tag fusion proteins for visualization or as a control in various biochemical experiments. It can also be used as a reporter gene to monitor the activity of a promoter or to track cell proliferation and differentiation.

eGFP is an enhanced version of GFP with a brighter fluorescent color and higher expression levels. It is a versatile protein with multiple applications and can be easily detected by UV light on a culture. GFP is typically tagged with an antibody or another protein to make it more specific for a biological function.

The chromophore of GFP is formed by the oxidation and cyclization of three amino acids in the center of a beta-barrel structure. When excited with UV light, GFP emits a bright green fluorescence. The high luminosity of GFP allows it to be easily visualized, even in complex cell structures or at low concentrations.

The EGFP molecular weight can be determined by SDS-PAGE, which involves running the protein on a polyacrylamide gel in a detergent such as sodium dodecyl sulfate. The protein is separated into parts, and the molecular weight is determined by comparing it to a standard ladder of known protein molecular weights.

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What is the eGFP Molecular Formula?

GFP is a protein that can be expressed in cells to produce a bright green fluorescence when excited with UV light. It was initially isolated from the jellyfish Aequorea victoria and has been used as a fluorescent marker for cells in many applications.

It is often paired with other fluorescent proteins to construct biosensors, e.g., a pair of biosensors with EGFP, and used to detect blood glucose levels or to visualize mRNA expression. In other applications, a single gene can be introduced into a cell to express eGFP in all cells or specific cell regions essential for a particular function. eGFP is also commonly used to determine if a transfection experiment was successful by shining UV light on the cell cultures and looking for the characteristic bright green glow.

An examination of electron density maps has identified the residues that differ between these two conformers. The difference is most pronounced for the side chain of the amino acid Leu18, which shifts between the two conformers with significant differences in the position and occupancy of the chromophore.

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