TOPIC: TRANSCRPTION PROCESS

IN EUKARYOTES
NAME: SUJI KEERTHAN B
 CERTIFICATE
CENTRAL BOARD OF SECONDARY EDUCATION

Certified that the Project Report Entitled

TRANSCRIPTION PROCESS IN EUKARYOTES

Is bonafide work carried out by SUJI KEERTHAN B in

fulfillment of he prescribed project work as instructed
by the secondary board of secondary education for
CBSE XII BIOLOGY during the academic year 2024-
25.The project report has been approved as it satisfies
the academic requirements as the project prescribed
for CBSE XII

INTERNAL EXTERNAL
 ACKNOWLEDGEMENT
I would like to express my special thanks of
gratitude to my BIOLOGY teacher Mrs.SEEMA
and Mr.RNR who helped me throughout this
BIOLOGY project.

Your guidance and encouragement have been

invaluable in helping me understand and
complete this project.

I am grateful for the opportunity on this

project and I am proud of the results I have
achieved . Thank you everyone for your
support and belief in my abilities
 OBJECTIVE
1.To study the process of transcription in eukaryotic
cells.
2.To understand the key components and stages
involved in the transcription process.
3.To explore the role of transcription factors, RNA
polymerase, and regulatory elements.
4. To analyze the differences between eukaryotic
and prokaryotic transcription.
 Materials
1. A microscope (for observing cells)
2. Laboratory slides
3. Eukaryotic cell culture (such as yeast or plant
cells)
4. Staining agents for DNA and RNA (e.g., DAPI,
Ethidium bromide)
5. RNA extraction kits
6. Gel electrophoresis apparatus
7. PCR machine (for amplification of transcription
products)
8. Reagents for RNA synthesis (e.g., RNA
polymerase, nucleotides, transcription factors)
 METHODS
1.Cell Preparation:
Cultivate eukaryotic cells (e.g., yeast, plant cells).
Prepare slides for microscopic observation of cells in
various stages of transcription.
2.DNA and RNA Extraction:
Extract DNA and RNA from the cultured cells using
an RNA extraction kit.
Use UV spectrophotometry to measure RNA quality
and quantity.
3.In Vitro Transcription Assay:
Set up in vitro transcription reactions using RNA
polymerase and cell extracts.
Add labeled nucleotides to monitor RNA synthesis
Introduction to Transcription in
Eukaryotes
Transcription is the first step in the process of
gene expression, where a segment of DNA is
copied into RNA by the enzyme RNA
polymerase. In eukaryotic cells, transcription
occurs in the nucleus and is a highly regulated
process that involves various proteins,
transcription factors, and the chromatin
structure.
Eukaryotic transcription is more complex than
in prokaryotes due to the presence of a
nucleus, multiple RNA polymerases, and a
variety of regulatory elements. The final
product of transcription is primarily messenger
RNA (mRNA), which is later translated into
proteins.
4.Gel Electrophoresis:
Separate RNA transcripts by size using agarose gel
electrophoresis.
Visualize RNA products under UV light.

5.Microscopic Observation:
Stain the cells and observe them under a microscope
to analyze transcription-related structures such as the
nucleolus and chromatin.

6.Reverse Transcription PCR (RT-PCR):Use RT-PCR to

convert mRNA to complementary DNA (cDNA) and
analyze specific transcriptional products.
Stages of Transcription
in Eukaryotes:
Eukaryotic transcription occurs in three main
stages: Initiation, Elongation, and Termination.
1.Initiation:

Chromatin Remodeling: Before transcription can

begin, chromatin needs to be relaxed (euchromatin)
so that RNA polymerase can access the DNA. This
involves histone modifications (e.g., acetylation).
Promoter Recognition: The process begins at the
promoter region of a gene. In eukaryotes, this
involves the TATA box, a conserved sequence found
in many genes.
Transcription Factors: General transcription factors
(TFs) such as TFIID, TFIIB, TFIIF, and TFIIH bind to the
promoter region and recruit RNA polymerase II.
Formation of the Pre-initiation Complex (PIC): The
combined transcription factors and RNA polymerase
II form a complex at the promoter.
2.Elongation:
RNA Polymerase II Activity: RNA polymerase moves
along the DNA template strand, synthesizing pre-
mRNA in the 5’ to 3’ direction. The polymerase
unwinds the DNA ahead of it and rewinds it behind.
Capping: The 5’ end of the pre-mRNA is capped with
a 7-methylguanosine cap, which protects the mRNA
from degradation and helps in translation initiation.
Splicing and Polyadenylation: As the RNA
polymerase synthesizes mRNA, splicing factors
remove introns and join exons together. The 3’ end
of the mRNA is also poly adenylated
3.Termination:

Cleavage and Release:

Once RNA polymerase reaches a termination signal,
the RNA is cleaved, and the RNA polymerase is
released from the DNA template . Post-transcriptional
Modifications: The newly synthesized mRNA undergoes
several modifications, including the addition of a poly-
A tail and the removal of introns (splicing)
Key Components Involved
in Transcription:
RNA Polymerase II : The enzyme responsible for
synthesizing mRNA in eukaryotes.
Promoter Region : A sequence of DNA where
transcription begins, often containing elements like the
TATA box.
Transcription Factors : Proteins that help regulate
transcription by binding to specific DNA sequences
near the gene being transcribed. Examples include
TFIID, TFIIB, and TFIIH.
Mediator Complex : A protein complex that bridges
the interaction between transcription factors and RNA
polymerase II.
Enhancers and Silencers : Regulatory DNA sequences
that can increase (enhancers) or decrease (silencers)
the rate of transcription.
Results and Observations:
Microscopic Observations : The nucleolus (where rRNA is
transcribed) is visible in cells actively transcribing ribosomal
RNA.Chromatin structure may appear condensed
(heterochromatin) or relaxed (euchromatin) depending on
transcription activity.

RNA Extraction Results:The total RNA extracted from cells

was analyzed for integrity using electrophoresis. A clear 28S
and 18S rRNA band indicates high-quality RNA.

Gel Electrophoresis:Bands corresponding to different

transcript lengths were observed in the gel, showing active
transcription of specific genes.The presence of a 5’ cap and
poly-A tail was confirmed by RT-PCR.
Discussion:
Regulation of Transcription: Eukaryotic
transcription is highly regulated. Specific
transcription factors, coactivators, and repressors
can fine-tune gene expression. The chromatin
structure also plays a crucial role, where tightly
packed chromatin (heterochromatin) is less
transcriptionally active than relaxed chromatin
(euchromatin).Differences from Prokaryotes:
Eukaryotes have multiple RNA polymerases (I, II,
III), whereas prokaryotes have only one. Eukaryotic
transcription occurs in the nucleus, whereas in
prokaryotes it occurs in the cytoplasm.
Additionally, eukaryotes require more transcription
factors and coactivators to regulate gene
expression.
Conclusion:
The transcription process in eukaryotes is intricate
and involves multiple steps, including chromatin
remodeling, promoter recognition, and RNA
processing. The proper functioning of transcription
factors, RNA polymerase, and associated proteins
ensures accurate gene expression. This project
highlights the complexity of gene regulation and the
importance of transcription in cellular processes..
 BIBLIOGRAPHY
wikipedia.org

www.genome.gov

www.nature.com