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#GFP

6 public questions tagged with this topic.

Genetics and Cell Biology

Which one of the following reporter genes can be used for real-time visualization of living cells/tissues in transgenic

In cell biology, cellular compartmentalization and cell cycle dictates that SNARE-mediated trafficking, Hayflick limit, P53 and cyclin regulation control proliferation. Therefore gfp is correct as it aligns with membrane trafficking and cell cycle control, supported by mechanistic studies and conserved across related systems.

Ref: Alberts et al., Molecular Biology of the Cell, 6th Edition, Chapter 17, Cell Cycle and Regulation, explains Cdc2, cyclins, P53, proteasomes, Hayflick limit, primary culture and immortalization mechanisms governing proliferation.

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Genetics and Cell Biology

Lysosomes of a cell were labelled with lysotracker Red. Subsequently, these cells were infected with GFP-transfected Myc

In cell biology, cellular compartmentalization and cell cycle dictates that SNARE-mediated trafficking, Hayflick limit, P53 and cyclin regulation control proliferation. Therefore GFP-Mycobacterium will be separated from lysotracker Red labeled lysosomes. is correct as it aligns with membrane trafficking and cell cycle control, supported by mechanistic studies and conserved across related

Ref: Alberts et al., Molecular Biology of the Cell, 6th Edition, Chapter 10, Membrane Structure and Transport, discusses SNARE proteins, vesicle trafficking, ER, Golgi, endosome membranes and selective transport providing compartmentalization foundation.

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Biochemistry

GFP, when overexpressed in a cell, remains mostly in the cytosol. A GFP construct is modified such that the resultant GF

Mechanistically, enzyme catalysis and lipid structure involves substrate affinity, transition state stabilization and phospholipid composition govern function. This validates nucleus because it reflects catalytic efficiency and membrane organization, a pattern repeatedly demonstrated in biochemistry research.

Ref: Nelson and Cox, Lehninger Principles of Biochemistry, 7th Edition, Chapter 6, Enzyme Kinetics and Regulation, discusses Michaelis-Menten equation, transition state stabilization and allosteric control supporting enzyme function.

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Blotting Techniques

GFP is used in blotting because:

Green fluorescent protein from jellyfish Aequorea victoria contains internal tripeptide Ser65-Tyr66-Gly67 that undergoes autocatalytic cyclization and oxidation to form p-hydroxybenzylidene-imidazolinone chromophore requiring no exogenous cofactor. When fused to protein of interest, chimera exhibits intrinsic fluorescence excitable at 395 and 475 nm, emitting at 509 nm, allowing direct visualization on gel documentation systems or after blotting using anti-GFP antibodies. Utility does not derive from heat emission, blocking ability, or binding inhibition. Self-sufficient fluorophore formation

Ref: NCERT Biology Class XII Principles on Klenow fill-in labeling, Lehninger Chapter 9 DNA cloning techniques, and Molecular Cloning by Sambrook Chapter 10 documenting end-labeling of cohesive termini.

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Microscopy

Why is GFP unique in detection?

Green fluorescent protein isolated from jellyfish Aequorea victoria contains intrinsic chromophore formed autocatalytically from tripeptide Ser-Tyr-Gly cyclization and oxidation within beta-barrel structure. No external cofactor, substrate or enzymatic activation is required beyond molecular oxygen for maturation. Once folded, it absorbs blue light near 395 nm and emits green photons near 509 nm. This self-contained fluorescence distinguishes it from luciferase which needs luciferin and ATP or peroxidase requiring luminol. Consequently GFP can be expressed as genetic fusion tag enabling live c

Ref: NCERT Biology Class XII Principles on Klenow fill-in labeling, Lehninger Chapter 9 DNA cloning techniques, and Molecular Cloning by Sambrook Chapter 10 documenting end-labeling of cohesive termini.

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Microscopy

Which microscopy is best to track protein movement in a living cell using GFP?

Green fluorescent protein fused to a protein of interest allows non-invasive tracking inside living cells, but requires a system that rejects background and enables time-lapse. Confocal microscopy uses laser excitation with pinhole filtering to capture sharp optical sections through living cytoplasm, minimizing photodamage compared to widefield while providing high signal-to-noise. Time-lapse acquisition of GFP dynamics reveals trafficking between endoplasmic reticulum, Golgi and plasma membrane, cytoskeletal assembly and nuclear import. Alternative modalities like bright field lack molecular

Ref: NCERT Biology Class XII Principles on Klenow fill-in labeling, Lehninger Chapter 9 DNA cloning techniques, and Molecular Cloning by Sambrook Chapter 10 documenting end-labeling of cohesive termini.

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