This document provides an overview of basic molecular biology concepts including:
1) DNA structure including nucleotides, base pairing, and the double helix formation.
2) Genes and genomes, including definitions of a gene, genome size comparisons, and that genes encode proteins.
3) The genetic code and mutations, including how the DNA sequence is translated into proteins and different types of mutations.
RNA Polymerase
Introduction
Purification
History
PRODUCTS OF RNAP
Messenger RNA
Non-coding RNA or "RNA genes
Transfer RNA
Ribosomal RNA
Micro RNA
Catalytic RNA (Ribozyme)
prokaryotic and eukaryotic
Transcription by RNA Polymerase
TYPES OF RNA POLYMERASE
Type I
Type II
Type III
Prokaryotic Transcription Unit
EXPRESSION OF A PROKARYOTIC GENE
Prokaryotic Polycistronic Message Codes for Several Different Proteins
Eukaryotic Transcription Unit
ENHANCERS AND SILENCERS
RESULT OF THE TRANSCRIPTION CYCLE
RNAP III TRANSCRIBES HUMAN MICRORNAS
RNAP I–specific subunits promotepolymerase clustering to enhance the rRNA genetranscription cycle
RNAP II–TFIIB STRUCTURE ANDMECHANISM OF TRANSCRIPTION INITIATION
FIVE CHECKPOINTS MAINTAINING THE FIDELITY OFTRANSCRIPTION BY RNAP IN STRUCTURAL ANDENERGETIC DETAILS
Replication Introduction , DNA replicating Models , Meselson and Stahl Experiments , Circuler Model of DNA replication , Replication in Prokaryotes , Replication In Eukaryotes , Comparison Between Prokaryotes and Eukaryotes Replicaton and PCR (Polymerease Chain Reaction)
RNA Polymerase
Introduction
Purification
History
PRODUCTS OF RNAP
Messenger RNA
Non-coding RNA or "RNA genes
Transfer RNA
Ribosomal RNA
Micro RNA
Catalytic RNA (Ribozyme)
prokaryotic and eukaryotic
Transcription by RNA Polymerase
TYPES OF RNA POLYMERASE
Type I
Type II
Type III
Prokaryotic Transcription Unit
EXPRESSION OF A PROKARYOTIC GENE
Prokaryotic Polycistronic Message Codes for Several Different Proteins
Eukaryotic Transcription Unit
ENHANCERS AND SILENCERS
RESULT OF THE TRANSCRIPTION CYCLE
RNAP III TRANSCRIBES HUMAN MICRORNAS
RNAP I–specific subunits promotepolymerase clustering to enhance the rRNA genetranscription cycle
RNAP II–TFIIB STRUCTURE ANDMECHANISM OF TRANSCRIPTION INITIATION
FIVE CHECKPOINTS MAINTAINING THE FIDELITY OFTRANSCRIPTION BY RNAP IN STRUCTURAL ANDENERGETIC DETAILS
Replication Introduction , DNA replicating Models , Meselson and Stahl Experiments , Circuler Model of DNA replication , Replication in Prokaryotes , Replication In Eukaryotes , Comparison Between Prokaryotes and Eukaryotes Replicaton and PCR (Polymerease Chain Reaction)
Cot curve dispersed repeated DNA or interspersed repeated DNA tandem repeated DNA Long interspersed repeat sequences (LINEs) Short interspersed nuclear elements (SINEs) satellite, minisatellite and microsatellite DNA Variable Number Tandem Repeat (or VNTR)
Dna supercoiling and role of topoisomerasesYashwanth B S
supercoiling is one of the important process to condenses the huge amount of DNA to fit inside the histone and its also plays a role during the replication ,transcription etc..,these activities is carried out by an enzyme called topoisomerases.
DNA supercoiling refers to the over- or under-winding of a DNA strand, and is an expression of the strain on that strand.Certain enzymes such as topoisomerases are able to change DNA topology to facilitate functions such as DNA replication or transcription
Genetic code, Deciphering of genetic code, properties of genetic code, Initiation & termination of codons, Gene Mutation, non sense codon, release factors, Transition , Trans versions
Cot curve dispersed repeated DNA or interspersed repeated DNA tandem repeated DNA Long interspersed repeat sequences (LINEs) Short interspersed nuclear elements (SINEs) satellite, minisatellite and microsatellite DNA Variable Number Tandem Repeat (or VNTR)
Dna supercoiling and role of topoisomerasesYashwanth B S
supercoiling is one of the important process to condenses the huge amount of DNA to fit inside the histone and its also plays a role during the replication ,transcription etc..,these activities is carried out by an enzyme called topoisomerases.
DNA supercoiling refers to the over- or under-winding of a DNA strand, and is an expression of the strain on that strand.Certain enzymes such as topoisomerases are able to change DNA topology to facilitate functions such as DNA replication or transcription
Genetic code, Deciphering of genetic code, properties of genetic code, Initiation & termination of codons, Gene Mutation, non sense codon, release factors, Transition , Trans versions
al-salam alykom ..
this lecture starts with basic definitions in genetic , also talk about DNA & RNA ( structures , types , similarities and differences ) .
it talks about bacterial DNA ( chromosome structure / plamids structure and functions / transponon types )
later , discusses about central dogma / gene expression starting from genetic code/codons , then DNA replication , trancription and finally translation
prepared by Sumia Abdalsalam Alfitoury / Libya
al-salam alykom ..
this lecture starts with basic definitions in genetic , also talk about DNA & RNA ( structures , types , similarities and differences ) .
it talks about bacterial DNA ( chromosome structure / plamids structure and functions / transponon types )
later , discusses about central dogma / gene expression starting from genetic code/codons , then DNA replication , trancription and finally translation
prepared by Sumia Abdalsalam Alfitoury / Libya
he culture media are classified in many different ways: Based on the physical state Liquid media Solid media Semisolid media Based on the presence or absence of oxygen Anaerobic media Aerobic media Based on nutritional factors Simple media Synthetic media Complex
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3. It’s all in the DNA
• Genetic material found in
every living cell
• Contains information to
make proteins
4. 1. DNA Structure & function
• Polymer made of 4 nucleotides : A, C, T, G
• polynucleotide eg. A-G-T-C-C-A-A-G-C-T-T….
• DNA is double stranded - Double-helix
• Complementary base pairing
• Anti-parallel
5. Nucleotides – the building blocks
of DNA
Nucleotide = Sugar + base + phosphate
Nucleoside = Sugar + base
Deoxyribose sugar 5 carbons
Phosphate (PO4) group at carbon no. 5
Hydroxyl (OH) group at carbon no. 3
www.mun.ca/biology/ scarr/Fg10_10rt.gif
10. Fig. 16-7
5 end
Hydrogen bond
3 end
1 nm
3.4 nm
3 end
0.34 nm
(a) Key features of DNA structure (b) Partial chemical structure
5 end
(c) Space-filling model
11. How one nucleotide can be joined to another through the 5’-PO4
(5 prime phosphate) and the 3’-OH (3 prime hydroxyl)
12. Many nucleotides can be joined in such a way to form a
polynucleotide chain a single stranded DNA
13. The Double Helix
The complete DNA molecule is made up of two complementary strands in
antiparallel directions
http://physicsweb.org/objects/world/16/3/7/pwhux4_03-03.jpg
14. 5’
3’
3’
5’
Complementary base-pairing
• A always pair with T
• C pairs with G
Antiparallel
• each strand of DNA has a “direction”
• at one end, the terminal carbon atom in
the backbone is the 5‟ carbon atom
• at the other end, the terminal carbon
atom is the 3‟ carbon atom
• therefore each DNA strand has a 5’ and a
3’ end
• in a double helix, the two strands are
always antiparallel
The length (or size) of a DNA molecule
is measured in basepairs (bp)
1 kilobase (kb) = 1,000 bp
1 Megabase (Mb) = 1,000,000 bp
15. Chromosomes and DNA
DNA is packaged in the
form of chromosomes in
the nucleus of a cell.
The chromosomes contain
DNA tightly wounded
around proteins.
16. 2. Genes and genomes
The Genome - complete set of DNA for an organism
A comparison of genomes
Organism
Amount of
DNA
No of
Chromosomes
No of genes
3 Gb
46
100,000
Yeast
13 Mb
16
6,000
E.coli
4.5 kb
1
1,000
Human
17. A Gene
• a specific DNA sequence that
contains genetic information
• information required to make a
specific type of protein
• that information is stored in the
sequence on the „sense‟ strand
• we say that a gene encodes a
protein
• thus a DNA molecule can contain
many genes
• the gene sequence is always
written 5‟ 3‟
5’ATGCTTGGACGTGATGACATTGGAGGA...
3’TACGAACCTGCACTACTGTAACCTCCT...
‘sense’
‘antisense’
18. 3. The Genetic Code and mutation
– how the DNA (or mRNA) sequence is translated into
the amino acid sequence of a protein
19. Reading frames
• There are 3 ways to read a gene sequence – reading frames
• Each reading frame will give a different result
• Only one reading frame is correct (usually)
20. Gene mutation
• Information coded in the DNA sequence is used to make proteins
• If the DNA sequence is changed, what will happen?
• A change in the genetic information is called a mutation. The outcome
depends on the nature of the „change‟.
• 3 types of DNA sequence mutations
21. Substitution mutations
-change of 1 base
-AAC ATA ACG CCG CGA GAT GAA –
Asn
Ile
Thr
Pro
Arg
Asp Glu
a. Silent mutation
- AAC ATC ACG CCG CGA GAT GAA –
b. Missense mutation
- AAC ATA AAG CCG CGA GAT GAA –
c. Nonsense mutation
- AAC ATA ACG CCG TGA GAT GAA –
What happens to
the amino acid
sequence?
What happens to
the protein?
23. Mutation can also happen due to
-changes in long DNA sequences
-changes in the structure of genes/ chromosomes
-changes in the number of genes/ chromosomes
-e.g. Mutations leading to cystic fibrosis
25. Denaturation – breaking up of the double helix molecule
•
•
•
•
•
hydrogen bonds broken
by heating to > 96oC (and also other conditions)
Tm = the melting point of a DNA molecule
depends on the GC content
higher %GC higher Tm
Renaturation - if the denatured DNA strands are allowed to
cool slowly
• the can re-anneal with each other and regain the double
helix structure
• reannealing by complementary base-pairing
26. 5. Comparative genomics
• Comparison of whole genome sequences
provides a highly detailed view of how
organisms are related to each other at the
genetic level. How are genomes compared
and what can these findings tell us about
how the overall structure of genes and
genomes have evolved?
• Purpose / benefits?