The document discusses the Green Fluorescent Protein (GFP), originally isolated from the jellyfish Aequorea victoria, detailing its structure, origin, working mechanism, and various applications in biotechnology. GFP is highlighted for its brightness, sensitivity, non-toxicity, and versatility in tracking protein localization, studying gene expression, and monitoring cellular activities. The document also includes multiple-choice questions and references related to GFP research.

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.

7. 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.”

8. 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.

10. 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.

11. • 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.

12. 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

13. • 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

14. 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.

15. • 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.

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

17. 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

18. MCQ Answers
1. Green Fluorescent protein
2. 25-27 KDa
3. Jelly fish
4. Aequorea victoria
5. Green

19. Reference
1. Chalfie M, Tu Y, Euskirchen G, Ward WW, Prasher
DC (Feb 1994). “Green fluorescent protein as a
marker for gene expression”. Science. 263 (5148):
802–5. Bibcode:1994Sci…263..802C.
Doi:10.1126/science.8303295. PMID 8303295. S2CID
904332
2. Barondeau DP, Kassmann CJ, Tainer JA, Getzoff ED
(Apr 2002). “Structural chemistry of a green
fluorescent protein Zn biosensor”. Journal of the
American Chemical Society. 124 (14): 3522–3524.
doi:10.1021/ja0176954. PMID 11929238.
3. Prendergast FG, Mann KG (Aug 1978). “Chemical and
physical properties of aequorin and the green
fluorescent protein isolated from Aequorea
forskålea”. Biochemistry. 17 (17): 3448–53.

20. 4. Yue JX, Holland ND, Holland LZ, Deheyn DD
(June 2016). “The evolution of genes encoding
for green fluorescent proteins: insights from
cephalochordates (amphioxus)”. Scientific
Reports. 6 (1): 28350.
Bibcode:2016NatSR…628350Y.
Doi:10.1038/srep28350. PMC 4911609. PMID
27311567.
5.Ormo M, Cubitt AB, Kallio K, Gross LA, Tsien
RY, Remington SJ. Crystal structure of the
Aequorea victoria green fluorescent protein.
Science. 1996;273:1392–1395. [PubMed] [Google
Scholar]

21. Thank
You…