GFP in rDNA Technology (Biotechnology).pptx

The green fluorescent protein (GFP) is a protein that exhibits green fluorescence when exposed to light in the blue to ultraviolet range.
The label GFP traditionally refers to the protein first isolated from the jellyfish Aequorea victoria and is sometimes called avGFP.
GFPs have been found in other organisms including Corals, sea anemones, zoanithids, copepods and lancelets.

1. YBT201 rDNA TECNOLOGY Topic: GFP Submitted to Ms. P. Mala
Assistant professor
Department of Biotechnology Submitted by Adithya V Biju 123011356005
First Year
Biotechnology

2. Contents.. Introduction Structure Origin Working Features Properties Application MCQ questions MCQ answers Reference

3. Introduction The green fluorescent protein (GFP) is a protein that exhibits green fluorescence when exposed to light in the blue to ultraviolet range. The label GFP traditionally refers to the protein first isolated from the jellyfish Aequorea victoria and is sometimes called avGFP . GFPs have been found in other organisms including Corals, sea anemones, zoanithids , copepods and lancelets.

4. Structure of the Aequorea victoria green fluorescent protein.

5. Structure of GFP It has 238 amino acid residues and a green fluorophore. Which is comprised of only three amino acids: Ser65-Tyr66-Gly67. Wild-type green fluorescent protein start out as a 238 amino acid peptide chain, about 25-27KDa with 11 β-sheets forming a cylindrical fence. Containing a chromophore in the center, preventing fluorescence quenching due to dipolarized water molecules or oxygen molecules.

6. Origin of GFP GFP is an endogenous protein from the jellyfish Aequorea Victoria. It was isolated by Osamu Shimomura in 1962. In 1992, the sequence of GFP was cloned Douglas Prasher and Martin Chalfie’s lab expressed the sequence in vivo. Roger Tsien’s lab improved GFP and managed to convert it into a commonly used research tool. In 2008, The Nobel Prize in Chemistry was awarded “for the discovery and development of the green fluorescent protein, GFP.”

7. Working of GFP Biologists use GFP as a marker protein. GFP can attach to and mark another protein with fluorescence, enabling scientists to see the presence of the particular protein in an organic structure. Gfp refers to the gene that produces green fluorescent protein.

8. Features of GFP Bright fluorescence: GFP has a bright green fluorescence that makes it easily visible under the right conditions.
High sensitivity: GFP is highly sensitive to changes in the environment, such as pH, temperature, and other factors, which makes it an important tool for monitoring cellular activity.
Non-toxic: GFP is non-toxic and does not interfere with cellular activity, making it a safe tool for studying cells and organisms.

9. Easy to use: GFP is easy to use and can be introduced into cells and organisms through genetic engineering.
Versatile: GFP is versatile and can be used in a wide range of applications, including studying gene expression, protein localization, and cellular behavior.
Stable: GFP is a stable protein that can be stored and used over a long period of time.
Safe: GFP is safe and does not cause any harm to cells or organisms, making it a valuable tool for studying the biology of living systems.

10. Properties of GFP Molecular weight: GFP has a molecular weight of approximately 27 kDa .
Fluorescence: GFP emits green light when exposed to ultraviolet or blue light. This fluorescence makes it an important tool for tracking cellular activity.
Absorption spectrum: GFP has a maximum absorption spectrum of 395 nm.
Emission spectrum: GFP has a maximum emission spectrum of 509 nm, which gives it its characteristic green fluorescence.
pH sensitivity: GFP is sensitive to changes in pH and its fluorescence can be affected by changes in the environment.

11. Temperature sensitivity: GFP is also sensitive to changes in temperature and its fluorescence can be affected by changes in the environment.
Stability: GFP is a relatively stable protein and can be stored for long periods of time without losing its fluorescence. Photostability : GFP is photostable and does not degrade quickly when exposed to light, making it an ideal tool for long-term studies.
Solubility: GFP is soluble in aqueous solutions, making it easy to work with in the laboratory.
Compatibility: GFP is compatible with a wide range of cells and organisms and can be used in a variety of experimental systems.

12. Application of GFP Tracking protein localization: GFP can be used to track the localization of proteins within cells, which provides insight into cellular processes and functions.
Studying gene expression: GFP can be used to study gene expression by fusing the GFP gene to a target gene of interest. This allows researchers to visualize the expression of the target gene within cells.

13. Monitoring cellular activity: GFP can be used to monitor cellular activity, including changes in pH, temperature, and other environmental factors, in real time.
Studying cellular behavior: GFP can be used to study cellular behavior, such as cell division, migration, and differentiation.
Genetic engineering: GFP can be used in genetic engineering to create new strains of organisms with specific traits.

14. MCQ Questions Full form GFP? a) Green Fluorescent Protein b)Grey Fluorescent protein c) Green fluro protein c) Grey fluro phosphate 2. Molecular weight of GFP? 34-37KDa b) 56-80KDa c) 25-27KDa d) 29-30KDa 3. GFP first isolated from? a) Rabbit b) Mouse c) Goat d) Jelly fish

15. 4. GFP produced by? a) Aequorea victoria b) Lion’s Mane c) Stygiomedusa gigantea d) Irukandji box 5. Which colour does GFP emit? a) Blue b) Green c) Grey d) Red

16. MCQ Answers Green Fluorescent protein 25-27 KDa Jelly fish Aequorea victoria Green

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