GREEN FLUORESCENT PROTIEN (GFP)

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GREEN FLUORESCENT PROTEIN (GFP) HARISH KUMAR GUGGILA

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Fluorescence: bright green colour to blue light Isolation: jellyfish Aequorea Victoria Use: reporter of expression INTRODUCTION GREEN FLUORESCENT PROTIEN (GFP)

GREEN FLUORESCENT PROTIEN (GFP) :

GREEN FLUORESCENT PROTIEN (GFP)

Structure of GFP::

Structure of GFP: Amino acid residues:238

Uses of new modified GFP: :

Uses of new modified GFP : To detect the weak promoter activity and in living single cells. Analysis of signal transduction pathways can be done. To visualize promoter activities and protein targeting into nucleus and different plastids.

BACKGROUND: :

BACKGROUND : GFP isolated from coelenterates which transduces blue chemiluminescence of the protein aequorin into green fluorescent light. The formulation of the fluorophore appears to be cell-autonomous apart from the requirement for molecular oxygen . In individual cells direct visualization of gene expression is possible without cell lysis and subsequent biochemical analysis. To minimise phototoxicity and photobleaching high intensity and more efficiency of expression is required under blue light. .

MATERIALS AND METHODS: Construction of plasmid is done. Protoplast transient expression should be known. Bombardment of tissues takes place. Transformation of tobacco should be done. Fluorescence microscopy. :

MATERIALS AND METHODS: Construction of plasmid is done . Protoplast transient expression should be known . Bombardment of tissues takes place . Transformation of tobacco should be done . Fluorescence microscopy.

RESULTS: ANALYSIS AND INTERPRETATION: :

RESULTS: ANALYSIS AND INTERPRETATION: Engineered GFP’s expression in plant cells: sGFP (S65T) as a reporter for a weak promoter sGFP (S65T) as a vital reporter in tobacco protoplasts: Analysis of organelle targeting in Arabidopsis: Nuclear targeting in onion epidermal cells: A favourable reporter for identification of transgenic plants: Drought-inducible gene expression in transgenic plants:

DISCUSSION: :

DISCUSSION: The usage of jellyfish GFP with high potential but in plants and mammals, its expression is shown inefficiently, formation of chromophore is slow, fluorescence is low and complex photoisomerization is seen. photobleaching and phototoxicity reduction modifications are made to improve GFP as versatile and sensitive reporter in several laboratories which allows simultaneous analysis of two promoters or two proteins and sensitivity of GFP detection increased. sGFP and sGFP(S65T) can be used to detect the activity of week promoters.

CONCLUSION: :

CONCLUSION: The new tools obtained can be used for studies of signal transduction, gene regulation development and cell biology in higher plants, where by using maize, tobacco, Arabidopsis cells sGFP and SGFP(65T) are sensitive reporters in gene expression .

Future development:

Future development Further developments of GFP provided variants with difference in emission and absorbance spectra which allows the simultaneous visualization of distinct GFP variants in cell. fluorescence imaging method.

THANK YOU:

THANK YOU