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COLLEGE OF AGRICULTURE INDORE
Credit seminar on Gene Expression
Guided By
:- Dr. Indu
Swarup
Submitted
by: -
Nitesh
Panwar
COLLEGE OF AGRICULTURE INDORE
Credit seminar on Gene Expression
Guided By
:- Dr. Indu
Swarup
Submitted
by: -
Nitesh
Panwar
GENE EXPRESSION
Gene Expression
DNA encodes hereditary information (genotype) ->
decoded into RNA -> protein (phenotype)
DNA
RNA
Protein
Transcription
Translation
LE 17-3-1
TRANSCRIPTION
DNA
Prokaryotic cell
LE 17-3-2
TRANSCRIPTION
DNA
Prokaryotic cell
Ribosome
Polypeptide
mRNA
Prokaryotic cell
LE 17-3-3
TRANSCRIPTION
TRANSLATION
DNA
mRNA
Ribosome
Polypeptide
DNA
Prokaryotic cell
Nuclear
envelope
TRANSCRIPTION
Eukaryotic cell
LE 17-3-4
TRANSCRIPTION
TRANSLATION
DNA
mRNA
Ribosome
Polypeptide
DNA
Pre-mRNA
Prokaryotic cell
Nuclear
envelope
mRNA
TRANSCRIPTION
RNA PROCESSING
Eukaryotic cell
LE 17-3-5
TRANSCRIPTION
TRANSLATION
DNA
mRNA
Ribosome
Polypeptide
DNA
Pre-mRNA
Prokaryotic cell
Nuclear
envelope
mRNA
TRANSLATION
TRANSCRIPTION
RNA PROCESSING
Ribosome
Polypeptide
Eukaryotic cell
• Main enzyme of this process is RNA polymerase
• RNA polymerase enzyme is a holo enzyme which is consist
of two units:- core enzyme and sigma factor
• Core enzyme is also consist of four sub units 2α, 1β, 1β’
• Sigma factor is activator of this process.
• Transcription unit is a part of DNA which is involved in
Transcription have three region Promoter, Structural
genes and Terminator
Transcription
LE 17-7
Promoter Transcription unit
RNA polymerase
Start point
DNA
5
3
3
5
Steps of transcription
1. Activation of Nucleotides
2. Attachment of RNA polymerase on DNA
3. Synthesis of RNA
4. Termination of Transcription
LE 17-7
Elongation
Non-template
strand of DNA
RNA
polymerase
RNA nucleotides
3 end
3
5
5
Newly made
RNA
Template
strand of DNA
Direction of transcription
(“downstream”)
Synthesis of an RNA Transcript
• The three stages of transcription:
– Initiation
– Elongation
– Termination
RNA processing in eukaryotes
1. Addition of methylated cap to 5’ end of
messenger RNA (mRNA)-> increases stability and
translation of mRNA
2. Addition of poly(A) tail to 3’ end (polyadenylation) ->
increases stability and translation of mRNA
3. Splicing
removal of introns and joining together of exons
All processing events occur in nucleus
before transport to cytoplasm
• RNA splicing:
carried out by spliceosomes
• Spliceosomes
complex of proteins and several small nuclear
ribonucleoproteins (snRNPs)
Recognize splice sites (specific RNA sequences)
cleave out introns and splice together exons (coding
region)
LE 17-11
Exon 1
5
Intron Exon 2
Other proteins
Protein
snRNA
snRNPs
RNA transcript (pre-mRNA)
Spliceosome
5
Spliceosome
components
Cut-out
intron
mRNA
Exon 1 Exon 2
5
TRANSLATION
• Translation- synthesis of polypeptide in
association with ribosome and directed by an
mRNA molucule with the help of tRNA and
several highly specific protein factor.
• or
• Translation is the process of decoding the
mRNA into a polypeptide chain
Translation process requires the
following major components
• 1. mRNA
• 2. tRNA
• 3. Ribosome (containing rRNA
• 4. Many other factors
MESSENGER RNA (mRNA)
• Carries the information for a specific protein
• Made up of 500 to 1000 nucleotides long
• Sequence of 3 bases called codon
• AUG – methionine or start codon
• UAA, UAG, or UGA – stop codons
TRANSFER RNA (tRNA)
• Clover-leaf shape
• Single stranded molecule with attachment site
at one end for an amino acid
• Opposite end has three nucleotide bases
called the anticodon
CLOVER LEAF STRUCTURE OF tRNA
amino acid
attachment site
U A C
anticodon
CODON AND ANTICODON
• The 3 bases of an anticodon are
complementary to the 3 bases of a codon
• Example: Codon AUG
Anticodon UAC
RIBOSOME
• Made of a large and small subunit
• Composed of rRNA (40%) and proteins (60%)
• Have two sites for tRNA attachment --- P and
A
RIBOSOME
• Large
sub unit
Small
Sub unit 26
P
Site
A
Site
mRNA
A U G C U A C U U C G
copyright cmassengale
Steps of translation
1. Activation of amino acids
2. Transfer of amino acid to t-RNA
3. Chain initiation
4. Elongation of polypeptide chain
5. Termination of polypeptide chain
6. Modification of released polypeptide
CHARGING OF tRNA
Amino acid reacts with ATP to form “Amino acyl
AMP enzyme complex”
This reaction catalyzed by a specific “Amino acyl
t-RNA snthetase” enzyme.
Amino acyl AMP-enzyme complex+t-RNA
=Amino acyl t-RNA complex +AMP +enzyme
Amino acyl t-RNA complex is called Charged t-
RNA
Now Amino acyl t-RNA moves to the ribosome
for protein synthesis.
THE TRANSLATION PROCESS
• The process of translation may be divided into
the following steps:
• 1. INITIATION
• 2.ELONGATION .
• 3.TERMINATION
STEP -1 INITIATION
• In this step 30 s and 50 s sub units of
ribosome ,GTP,mg+2 ,charged t-RNA ,m-
RNA and some initiation factor are
required.
• In prokaryotes there are three initiation
factor present –IF1,IF2,IF3
• In Eukaryotes ten initiation factors have
been identified in red blood cells-
• eIF1, eIF2,
eIF3,eIF4A,eIF4B,eIF4C,eIF4D,eIF4F,eI
F5,eIF6
32
INITIATION
mRNA
A U G C U A C U U C G
2-tRNA
G
aa2
A U
A
1-tRNA
U A C
aa1
anticodon
hydrogen
bonds codon
copyright cmassengale
ELONGATION
• New tRNA with new amino acid ia attach at ‘A’
site of ribosome.
• The link between amino acid of ‘P’ site of tRNA is
broken and t-RNA of P-site is discharged so -
COOH of P-site A.A. Becomes free.
• Now peptide bond takes place between –COOH
group of P-site amino acid and –NH2 group of A-
site amino site.
• Now t-RNA OF A- site is transferred to P-site and
A-site become empty.
35
mRNA
A U G C U A C U U C G
1-tRNA 2-tRNA
U A C G
aa1 aa2
A U
A
anticodon
hydrogen
bonds codon
peptide bond
3-tRNA
G A A
aa3
Elongation
copyright cmassengale
36
mRNA
A U G C U A C U U C G
1-tRNA
2-tRNA
U A C
G
aa1
aa2
A U
A
peptide bond
3-tRNA
G A A
aa3
Ribosomes move over one codon
(leaves)
copyright cmassengale
37
mRNA
A U G C U A C U U C G
2-tRNA
G
aa1
aa2
A U
A
peptide bonds
3-tRNA
G A A
aa3
4-tRNA
G C U
aa4
A C U
copyright cmassengale
38
mRNA
G C U A C U U C G
aa1
aa2
A
peptide bonds
3-tRNA
G A A
aa3
4-tRNA
G C U
aa4
A C U
U G A
5-tRNA
aa5
copyright cmassengale
39
mRNA
G C U A C U U C G
aa1
aa2
A
peptide bonds
3-tRNA
G A A
aa3
4-tRNA
G C U
aa4
A C U
U G A
5-tRNA
aa5
Ribosomes move over one codoncopyright cmassengale
TERMINATION
• Due to slidind of ribosome over m-RNA when any
Nonsence codon (UUA,UAG,UGA)available at A-
site of ribosome ,then polypeptide chain
terminates.
• The linkage between the last t-RNA and the
polypeptide chain is broken by three release
factors called RF1, RF2, RF3 with the help of GTP.
• In eukaryotes only one release factor is known –
eRF1.
42
mRNA
A C A U G U
aa1
aa2
U
primary
structure
of a protein
aa3
200-tRNA
aa4
U A G
aa5
C U
aa200
aa199
terminator
or stop
codon
Termination
copyright cmassengale
43
End Product –The Protein
• The end products of protein synthesis is
a primary structure of a protein
• A sequence of amino acid bonded
together by peptide bonds
aa1
aa2 aa3 aa4
aa5
aa200
aa199
copyright cmassengale
THANK YOU

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GIÁO ÁN DẠY THÊM (KẾ HOẠCH BÀI BUỔI 2) - TIẾNG ANH 8 GLOBAL SUCCESS (2 CỘT) N...
 

Protein Synthesis

  • 1. COLLEGE OF AGRICULTURE INDORE Credit seminar on Gene Expression Guided By :- Dr. Indu Swarup Submitted by: - Nitesh Panwar
  • 2. COLLEGE OF AGRICULTURE INDORE Credit seminar on Gene Expression Guided By :- Dr. Indu Swarup Submitted by: - Nitesh Panwar GENE EXPRESSION
  • 3. Gene Expression DNA encodes hereditary information (genotype) -> decoded into RNA -> protein (phenotype) DNA RNA Protein Transcription Translation
  • 9. • Main enzyme of this process is RNA polymerase • RNA polymerase enzyme is a holo enzyme which is consist of two units:- core enzyme and sigma factor • Core enzyme is also consist of four sub units 2α, 1β, 1β’ • Sigma factor is activator of this process. • Transcription unit is a part of DNA which is involved in Transcription have three region Promoter, Structural genes and Terminator Transcription
  • 10. LE 17-7 Promoter Transcription unit RNA polymerase Start point DNA 5 3 3 5
  • 11. Steps of transcription 1. Activation of Nucleotides 2. Attachment of RNA polymerase on DNA 3. Synthesis of RNA 4. Termination of Transcription
  • 12. LE 17-7 Elongation Non-template strand of DNA RNA polymerase RNA nucleotides 3 end 3 5 5 Newly made RNA Template strand of DNA Direction of transcription (“downstream”)
  • 13. Synthesis of an RNA Transcript • The three stages of transcription: – Initiation – Elongation – Termination
  • 14.
  • 15. RNA processing in eukaryotes 1. Addition of methylated cap to 5’ end of messenger RNA (mRNA)-> increases stability and translation of mRNA 2. Addition of poly(A) tail to 3’ end (polyadenylation) -> increases stability and translation of mRNA 3. Splicing removal of introns and joining together of exons All processing events occur in nucleus before transport to cytoplasm
  • 16. • RNA splicing: carried out by spliceosomes • Spliceosomes complex of proteins and several small nuclear ribonucleoproteins (snRNPs) Recognize splice sites (specific RNA sequences) cleave out introns and splice together exons (coding region)
  • 17.
  • 18. LE 17-11 Exon 1 5 Intron Exon 2 Other proteins Protein snRNA snRNPs RNA transcript (pre-mRNA) Spliceosome 5 Spliceosome components Cut-out intron mRNA Exon 1 Exon 2 5
  • 19. TRANSLATION • Translation- synthesis of polypeptide in association with ribosome and directed by an mRNA molucule with the help of tRNA and several highly specific protein factor. • or • Translation is the process of decoding the mRNA into a polypeptide chain
  • 20. Translation process requires the following major components • 1. mRNA • 2. tRNA • 3. Ribosome (containing rRNA • 4. Many other factors
  • 21. MESSENGER RNA (mRNA) • Carries the information for a specific protein • Made up of 500 to 1000 nucleotides long • Sequence of 3 bases called codon • AUG – methionine or start codon • UAA, UAG, or UGA – stop codons
  • 22. TRANSFER RNA (tRNA) • Clover-leaf shape • Single stranded molecule with attachment site at one end for an amino acid • Opposite end has three nucleotide bases called the anticodon
  • 23. CLOVER LEAF STRUCTURE OF tRNA amino acid attachment site U A C anticodon
  • 24. CODON AND ANTICODON • The 3 bases of an anticodon are complementary to the 3 bases of a codon • Example: Codon AUG Anticodon UAC
  • 25. RIBOSOME • Made of a large and small subunit • Composed of rRNA (40%) and proteins (60%) • Have two sites for tRNA attachment --- P and A
  • 26. RIBOSOME • Large sub unit Small Sub unit 26 P Site A Site mRNA A U G C U A C U U C G copyright cmassengale
  • 27. Steps of translation 1. Activation of amino acids 2. Transfer of amino acid to t-RNA 3. Chain initiation 4. Elongation of polypeptide chain 5. Termination of polypeptide chain 6. Modification of released polypeptide
  • 28. CHARGING OF tRNA Amino acid reacts with ATP to form “Amino acyl AMP enzyme complex” This reaction catalyzed by a specific “Amino acyl t-RNA snthetase” enzyme. Amino acyl AMP-enzyme complex+t-RNA =Amino acyl t-RNA complex +AMP +enzyme Amino acyl t-RNA complex is called Charged t- RNA Now Amino acyl t-RNA moves to the ribosome for protein synthesis.
  • 29. THE TRANSLATION PROCESS • The process of translation may be divided into the following steps: • 1. INITIATION • 2.ELONGATION . • 3.TERMINATION
  • 30. STEP -1 INITIATION • In this step 30 s and 50 s sub units of ribosome ,GTP,mg+2 ,charged t-RNA ,m- RNA and some initiation factor are required. • In prokaryotes there are three initiation factor present –IF1,IF2,IF3 • In Eukaryotes ten initiation factors have been identified in red blood cells- • eIF1, eIF2, eIF3,eIF4A,eIF4B,eIF4C,eIF4D,eIF4F,eI F5,eIF6
  • 31.
  • 32. 32 INITIATION mRNA A U G C U A C U U C G 2-tRNA G aa2 A U A 1-tRNA U A C aa1 anticodon hydrogen bonds codon copyright cmassengale
  • 33. ELONGATION • New tRNA with new amino acid ia attach at ‘A’ site of ribosome. • The link between amino acid of ‘P’ site of tRNA is broken and t-RNA of P-site is discharged so - COOH of P-site A.A. Becomes free. • Now peptide bond takes place between –COOH group of P-site amino acid and –NH2 group of A- site amino site. • Now t-RNA OF A- site is transferred to P-site and A-site become empty.
  • 34.
  • 35. 35 mRNA A U G C U A C U U C G 1-tRNA 2-tRNA U A C G aa1 aa2 A U A anticodon hydrogen bonds codon peptide bond 3-tRNA G A A aa3 Elongation copyright cmassengale
  • 36. 36 mRNA A U G C U A C U U C G 1-tRNA 2-tRNA U A C G aa1 aa2 A U A peptide bond 3-tRNA G A A aa3 Ribosomes move over one codon (leaves) copyright cmassengale
  • 37. 37 mRNA A U G C U A C U U C G 2-tRNA G aa1 aa2 A U A peptide bonds 3-tRNA G A A aa3 4-tRNA G C U aa4 A C U copyright cmassengale
  • 38. 38 mRNA G C U A C U U C G aa1 aa2 A peptide bonds 3-tRNA G A A aa3 4-tRNA G C U aa4 A C U U G A 5-tRNA aa5 copyright cmassengale
  • 39. 39 mRNA G C U A C U U C G aa1 aa2 A peptide bonds 3-tRNA G A A aa3 4-tRNA G C U aa4 A C U U G A 5-tRNA aa5 Ribosomes move over one codoncopyright cmassengale
  • 40. TERMINATION • Due to slidind of ribosome over m-RNA when any Nonsence codon (UUA,UAG,UGA)available at A- site of ribosome ,then polypeptide chain terminates. • The linkage between the last t-RNA and the polypeptide chain is broken by three release factors called RF1, RF2, RF3 with the help of GTP. • In eukaryotes only one release factor is known – eRF1.
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  • 42. 42 mRNA A C A U G U aa1 aa2 U primary structure of a protein aa3 200-tRNA aa4 U A G aa5 C U aa200 aa199 terminator or stop codon Termination copyright cmassengale
  • 43. 43 End Product –The Protein • The end products of protein synthesis is a primary structure of a protein • A sequence of amino acid bonded together by peptide bonds aa1 aa2 aa3 aa4 aa5 aa200 aa199 copyright cmassengale