Tuesday, February 18, 2020
GFP Mutants with Altered Mass Spectra Research Proposal - 1
GFP Mutants with Altered Mass Spectra - Research Proposal Example The green fluorescent protein (GFP), was discovered and isolated from the from the jellyfish Aequorea victoria (1). The native protein contains 238 amino acids which fold into six alpha helices and eleven beta strands forming a classical cylindrical beta sheet with anti-parallel strands. The GFP protein owes its fluorescence to the derived chromophore that is buried deep in the beta-barrel which protects interactions with the solvent. Due to its stable fluorescence, GFP has been used in biological studies as an important marker for processes like protein trafficking. Sequence data for GFP has led to the development of recombinant GFP proteins with modifications to increase the fluorescence intensity and longevity. There are many mutant GFPs with differences in absorption and emission spectra. There are enhanced GFP (EGFP) that has higher-intensity emission after blue-light excitation compared to wild-type GFP (4) (3). Another is GFPuv, a recombinant GFP with maximum emission spectra is 509nm, similar to that of wild-type GFP. GFPuv has a molecular weight of 29 kDa with three amino acid substitutions (Phe-99 to Ser, Met-153 to Thr, and Val-163 to Ala resulting in 18 times more brightness under ultra-violet (UV) light at 395nm. GFPuv is 18 times brighter than wild-type GFP. The large fluorescence of GFPuv making it suitable for experiments that require detection of changes in fluorescence. GFPuv expressed in highly expressed in E. coli is a soluble, fluorescent protein in contrast to wild-type GFP, which may be expressed in inclusion bodies as a nonfluorescent protein.