GFP Green fluorescent protein - Essay Example

Key Applications of GFP in Fundamental Cell and Molecular Biology Research Green Fluorescent Protein molecule is considered to be an expedient fluorescent protein. It is a boon to present research studies to understand the fundamental biology of cell and molecules. The fluorescent properties of GFP molecule enable researchers to understand the gene expression in diverse animal models. GFP molecule has an assemblage of amino acids which is built in light managing apparatus, the chromophore which is ideal for genetic engineering experiments.

A range of research studies engage GFP as a reporter molecule in order to understand the functions of molecules and genes explicitly (Chalfie, 1994). GFP as Reporter Molecule Microorganisms can be easily used for genetic tailoring by means of genetic tools to study the modifications in environmental conditions. Reporter genes are tagged with multicoloured fluorescent proteins which aid in identifying the chemical effects prevailing in the environment. The engineered microorganisms are then used for whole cell array set-up on silicon chips and diverse optic fibres to get bioavailability of pollutants which could be correlated with their impact on life (Belkin, 2003).

Reporter gene is expressed when it is fused with a promoter target sequences. Thus expression can be observed by the appearance of the reporter gene, in view of the fact that reporter is tagged with the active gene of interest. The promoter is tagged with a marker gene such as GFP, a stable molecule used to identify the phenotype or the genotype of the transformed organism. Luminescence markers: emit light making the selection process easy and simple moreover, they measure the cellular activity swiftly.

FACS (fluorescence activated cell sorting) is performed with GFP, YFP and BFP i.e. green, yellow and blue fluorescent proteins respectively. The molecules do not require oxygen for fluorescence (Belkin, 2003).In transgenic organisms GFP molecule is integrated with active gene, expression of gene is observed through green glow under UV radiation. Transgenic organisms of various species such as E coli, C elegans, Drosophila were made to glow by inserting GFP reporter gene. Toxicity of GFP is not observed in both prokaryotic and eukaryotic organism, indicating the safety of the molecule.

Small size of GFP makes it ideal for reporter gene or if GFP is tagged at the end of the active gene, it is expressed with the gene once the gene undergoes the process of transcription and translation, however, expression of GFP does not affect the protein; conversely, the protein can be seen as green fluorescence indicating the gene expression. All these properties of GFP make it an ideal for observing real-time events inside a cell, through fluorescent resonance energy transfer (FRET). This assists in understanding intracellular molecular interactions, drug-receptor interaction or substrate-enzyme interaction (Miyawaki, 2004.). JAK/STAT signaling plays vital role in development of Drosophila, tagging GFP enables one to investigate role of JAK/STAT signaling in Drosophila (Bach et al., 2006). GFP can be used to work as biosensors by tagging GFP with the heat shock protein (hsp) expressing gene, hsp-GFP.

Drosophila and other microbial organisms can be utilized for pollution monitoring because their genomes can be tailored. The heat shock proteins (hsp) or the stress sensing proteins are expressed in response to small changes in the microenvironment. Tagging reporter with hsps such as GFP will be reported by fluorescence, which is easily detected. Integration of two reporter genes within same organism is called dual labeling. In the process, one reporter acts as an internal control and other responder to analyze, such as incorporation of GFP and YFP in E.

coli, making construct as recA’::egfp and grpE’::dsRedExpress. The recA promoter responds to DNA damage grpE is heat shock protein, activated against the stress induced by chemical agents combination helps in monitoring of genotoxicity and normal toxicity (Belkin, 2003).ReferencesBach, E. A., Ekas, L. A., Ayala-Camargo, A., Flaherty, M. S., et al. (2006). GFP reporters detectthe activation of the Drosophila JAK/STAT pathway in vivo. Gene Expression Patterns,7(3), 323-31. Belkin, S., (2003).

Microbial whole-cell sensing systems of environmental pollutants. Current Opinion in Microbiology, 6, 206- 212.Chalfie, M., Tu, Y., Euskirchen, G., Ward, W. W. and Prasher, D. C. (1994). Green fluorescent protein as a marker for gene expression. Science, 263, 802-805.Miyawaki, A. (2004). Cell Structure and Function, 29, 6.