BACTERIAL GENETICS

10.1 Experiments showed that DNA is the genetic material

Frederick Griffith discovered that a transforming factor could be transferred into a bacterial cell
Disease-causing bacteria were killed by heat Harmless bacteria were incubated with heat-killed bacteria Some harmless cells were converted to diseasecausing bacteria, a process called transformation The disease-causing characteristic was inherited by descendants of the transformed cells

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10.1 Experiments showed that DNA is the genetic material

Alfred Hershey and Martha Chase used bacteriophages to show that DNA is the genetic material
Bacteriophages are viruses that infect bacterial cells
Phages were labeled with radioactive sulfur to detect proteins or radioactive phosphorus to detect DNA

Bacteria were infected with either type of labeled phage to determine which substance was injected into cells and which remained outside

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10.1 Experiments showed that DNA is the genetic material

The sulfur-labeled protein stayed with the phages outside the bacterial cell, while the phosphorus-labeled DNA was detected inside cells

Cells with phosphorus-labeled DNA produced new bacteriophages with radioactivity in DNA but not in protein

Animation: Hershey-Chase Experiment

Animation: Phage T2 Reproductive Cycle

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Head DNA Tail Tail fiber

Head
DNA Tail Tail fiber

Phage Bacterium

Radioactive protein

Empty protein shell Phage DNA

Radioactivity in liquid

DNA Batch 1 Radioactive protein

Centrifuge Pellet

1 Mix radioactively

2 Agitate in a blender to

3 Centrifuge the mixture

4 Measure the

labeled phages with bacteria. The phages infect the bacterial cells.

separate phages outside the bacteria from the cells and their contents.

so bacteria form a pellet at the bottom of the test tube.

radioactivity in the pellet and the liquid.

Batch 2 Radioactive DNA

Radioactive DNA Centrifuge Pellet

Radioactivity in pellet

Phage Bacterium

Radioactive protein

Empty protein shell Phage DNA

DNA Batch 1 Radioactive protein

1 Mix radioactively

2 Agitate in a blender to

labeled phages with bacteria. The phages infect the bacterial cells.

separate phages outside the bacteria from the cells and their contents.

Batch 2 Radioactive DNA

Radioactive DNA

Empty protein shell Phage DNA

Radioactivity in liquid

Centrifuge Pellet
3 Centrifuge the mixture 4 Measure the

so bacteria form a pellet at the bottom of the test tube.

radioactivity in the pellet and the liquid.

Centrifuge Pellet Radioactivity in pellet

Phage attaches to bacterial cell.

Phage injects DNA.

Phage DNA directs host cell to make more phage DNA and protein parts. New phages assemble. Cell lyses and releases new phages.

THE STRUCTURE OF THE GENETIC MATERIAL

Nucleic Acids
DNA and RNA

What we already know:

The nucleus contains DNA Eukaryotes have linear DNA

Prokaryotes have circular DNA

During cell division/growth, the DNA is copied and one copy is transferred to progeny

What we already know:

Traits are distinguishing features or characteristics Traits are passed from parent to offspring Traits are passed on through genes Genes are specific sections of DNA In sexual reproduction, offspring get half of their genes from each parent

DNA is INFORMATION
DNA= deoxyribose nucleic acid A molecule that stores information

Contains the instructions for making proteins Like a cookbook contains the instructions for making a cake

Hundreds of thousands of proteins exist inside each one of us to help carry out our daily functions. These proteins are produced locally, assembled piece-by-piece to exact specifications based on the code carried in the DNA

Nucleic Acids DNA and RNA

DNA - deoxyribonucleic acid RNA - ribonucleic acid

DNA- stores genetic information and passes it to the next generation RNA - use the coded information of DNA in protein synthesis

DNA and RNA are polymers of nucleotides

Proteins are polypeptides, Carbohydrates are polysaccharides

Nucleic acids are polynucleotides

The monomer unit of DNA and RNA is the nucleotide, containing

Nitrogenous base

5-carbon sugar
Phosphate group
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Nucleoside Nucleotide

The Structure of Nucleic Acids

DNA and RNA are polymers called polynucleotides
A sugar-phosphate backbone is formed by covalent bonding between the phosphate of one nucleotide and the sugar of the next nucleotide Nitrogenous bases extend from the sugar-phosphate backbone

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Animation: DNA and RNA Structure

The primary structure of DNA is the sequence

5 end

3 Phosphodiester linkage

3 end

Sugar-phosphate backbone Phosphate group Nitrogenous base Sugar Nitrogenous base (A, G, C, or T) DNA nucleotide Phosphate group

Thymine (T)

Sugar (deoxyribose)
DNA nucleotide

DNA polynucleotide

Different pentose sugars in RNA & DNA RNA

Sugar carbons have prime numbers, to distinguish them from atoms in bases

DNA

Nucleotides
Deoxyribonucleotides

Ribonucleotides

Heterocyclic Nitrogen Bases

RNA

DNA

The secondary structure of DNA is the double helix

25

DNA is a double-stranded helix

James D. Watson and Francis Crick deduced the secondary structure of DNA, with X-ray crystallography data from Rosalind Franklin and Maurice Wilkins DNA is composed of two polynucleotide chains joined together by hydrogen bonding between bases, twisted into a helical shape The sugar-phosphate backbone is on the outside The nitrogenous bases are perpendicular to the backbone in the interior Specific pairs of bases give the helix a uniform shape A pairs with T, forming two hydrogen bonds

G pairs with C, forming three hydrogen bonds

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The secondary structure of DNA

Two anti-parallel polynucleotide chains wound around the same axis. Sugar-phosphate chains wrap around the periphery. Bases (A, T, C and G) occupy the core, forming complementary A T and G C Watson-Crick base pairs. 27

5 end P
4 3 5 2 1

3 end
2 3 1 4 5

P
3 end 5 end

The DNA double helix is held together

mainly by- Hydrogen bonds

hydrogen bonding;
base stacking

29

Hydrogen bond
a chemical bond in which a hydrogen atom of one molecule is attracted to an electronegative atom, especially a nitrogen, oxygen, or fluorine atom, usually of another molecule.

30

Two hydrogen bonds between A:T pairs Three hydrogen bonds between C: G paire 31

Base Stacking The bases in DNA are

planar and have a

tendency to "stack". Major stacking forces: hydrophobic interaction van der Waals forces.
32

In eukaryotic cells,
DNA is folded into chromatin

DNA Tertiary Structure

DNA DOUBLE HELICAL STRUCTURE COILS ROUND HISTONES.
DNA BOUND TO HISTONES FORMS NUCLEOSOMES (10nm FIBRES) NUCLEOSOMES CONTAIN 146 NUCLEOTIDES

Nucleosomes
any of the repeating globular subunits of chromatin that consist of a complex of DNA and histone

Supercoil = coil over coil

Supercoil = coil over coil

Biochemistry for

37

Functions of DNA and summary of structure

DNA consists of four basesA, G, C, and Tthat are held in linear array by phosphodiester bonds through the 3' and 5' positions of adjacent deoxyribose moieties. DNA is organized into two strands by the pairing of bases A to T and G to C on complementary strands. These strands form a double helix around a central axis. The 3 x 109 base pairs of DNA in humans are organized into the haploid complement of 23 chromosomes. DNA provides a template for its own replication and thus maintenance of the genotype and for the transcription of the roughly 30,000 human genes into a variety of RNA molecules.

DNA REPLICATION

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DNA replication depends on specific base pairing

DNA replication follows a semiconservative model
The two DNA strands separate Each strand is used as a pattern to produce a complementary strand, using specific base pairing

Each new DNA helix has one old strand with one new strand

Animation: DNA Replication Overview

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Parental molecule of DNA

Nucleotides Parental molecule of DNA Both parental strands serve as templates

Nucleotides Parental molecule of DNA Both parental strands serve as templates Two identical daughter molecules of DNA

THE FLOW OF GENETIC INFORMATION FROM DNA TO RNA TO PROTEIN

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10.6 The DNA genotype is expressed as proteins, which provide the molecular basis for phenotypic traits
A gene is a sequence of DNA that directs the synthesis of a specific protein (genotype) DNA is transcribed into RNA RNA is translated into protein The presence and action of proteins determine the phenotype of an organism

The DNA genotype is expressed as proteins, which provide the molecular basis for phenotypic traits
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Genotype Vs Phenotype
Genotype: it is the genetic constitution of an organism/individual. The set of genes that it carries and this effects the phenotype of an individual . These inherited instructions may or may not cause an observable effect on the individual.

Phenotype: It is an observable characteristic of an organisms/individual (as morphology, development, behavious etc.) which is influenced both by its genotype and by the environment.

DNA

Nucleus Cytoplasm

DNA Transcription

RNA
Nucleus Cytoplasm

DNA Transcription

RNA
Nucleus Cytoplasm Translation

Protein

Genetic information written in codons is translated into amino acid sequences

The sequence of nucleotides in DNA provides a code for constructing a protein
Protein construction requires a conversion of a nucleotide sequence to an amino acid sequence Transcription rewrites the DNA code into RNA, using the same nucleotide language Each word is a codon, consisting of three nucleotides Translation involves switching from the nucleotide language to amino acid language Each amino acid is specified by a codon
64 codons are possible Some amino acids have more than one possible codon
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DNA molecule Gene 1

Gene 2

Gene 3

DNA strand Transcription RNA Codon Translation Polypeptide Amino acid

DNA strand

Transcription RNA Codon Translation

Polypeptide
Amino acid

The genetic code is the Rosetta stone of life

Characteristics of the genetic code
Triplet: Three nucleotides specify one amino acid
61 codons correspond to amino acids AUG codes for methionine and signals the start of transcription 3 stop codons signal the end of translation

First base

Second base

Third base

The genetic code is the Rosetta stone of life

Redundant: More than one codon for some amino acids Unambiguous: Any codon for one amino acid does not code for any other amino acid Does not contain spacers or punctuation: Codons are adjacent to each other with no gaps in between Nearly universal

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First base

Second base

Third base

Strand to be transcribed

DNA

Strand to be transcribed

DNA

Transcription

RNA Start codon Stop codon

Strand to be transcribed

DNA

Transcription

RNA Start codon Stop codon

Translation

Polypeptide

Met

Lys

Phe

Review: The flow of genetic information in the cell is DNA RNA protein
Does translation represent:
DNA RNA or RNA protein?

Where does the information for producing a protein originate:

DNA or RNA?

Which one has a linear sequence of codons:

rRNA, mRNA, or tRNA?

Which one directly influences the phenotype:

DNA, RNA, or protein?
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DNA

Transcription

mRNA

RNA polymerase Translation

1 mRNA is transcribed from a DNA template.

Amino acid

Enzyme

2 Each amino acid attaches to its proper tRNA with the help of a specific enzyme and ATP.

ATP tRNA

Anticodon Initiator tRNA Large ribosomal subunit

3 Initiation of polypeptide synthesis The mRNA, the first tRNA, and the ribosomal sub-units come together.

Start Codon mRNA

Small ribosomal subunit

Growing polypeptide

New peptide bond forming

Codons mRNA

4 Elongation A succession of tRNAs add their amino acids to the polypeptide chain as the mRNA is moved through the ribosome, one codon at a time.

Polypeptide

Stop codon

5 Termination The ribosome recognizes a stop codon. The polypeptide is terminated and released.

DNA

Transcription

mRNA RNA polymerase Amino acid Translation

1 mRNA is transcribed from a DNA template.

2 Each amino acid

Enzyme

attaches to its proper tRNA with the help of a specific enzyme and ATP.

ATP tRNA

Anticodon Initiator tRNA Large ribosomal subunit

3 Initiation of polypeptide synthesis

The mRNA, the first tRNA, and the ribosomal sub-units come together. Start Codon mRNA

Small ribosomal subunit

Growing polypeptide

New peptide bond forming

4 Elongation

Codons mRNA

A succession of tRNAs add their amino acids to the polypeptide chain as the mRNA is moved through the ribosome, one codon at a time.

Polypeptide

5 Termination

Stop codon

The ribosome recognizes a stop codon. The polypeptide is terminated and released.

Mutations can change the meaning of genes

A mutation is a change in the nucleotide sequence of DNA
Base substitutions: replacement of one nucleotide with another
Effect depends on whether there is an amino acid change that alters the function of the protein

Deletions or insertions
Alter the reading frame of the mRNA, so that nucleotides are grouped into different codons
Lead to significant changes in amino acid sequence downstream of mutation

Cause a nonfunctional polypeptide to be produced

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10.16 Mutations can change the meaning of genes

Mutations can be
Spontaneous: due to errors in DNA replication or recombination Induced by mutagens
High-energy radiation Chemicals

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Normal hemoglobin DNA

Mutant hemoglobin DNA

mRNA

mRNA

Normal hemoglobin

Sickle-cell hemoglobin

Glu

Val

Normal gene mRNA Protein

Met

Lys

Phe

Gly

Ala

Base substitution

Met

Lys

Phe

Ser

Ala

Base deletion

Missing

Met

Lys

Leu

Ala

His

MICROBIAL GENETICS

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Bacteria can transfer DNA in three ways

Three mechanisms allow transfer of bacterial DNA
Transformation is the uptake of DNA from the surrounding environment Transduction is gene transfer through bacteriophages Conjugation is the transfer of DNA from a donor to a recipient bacterial cell through a cytoplasmic bridge

Recombination of the transferred DNA with the host bacterial chromosome leads to new combinations of genes
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DNA enters cell

Fragment of DNA from another bacterial cell Bacterial chromosome (DNA)

Phage

Fragment of DNA from another bacterial cell (former phage host)

Mating bridge

Sex pili

Donor cell (male)

Recipient cell (female)

Donated DNA

Crossovers

Degraded DNA

Recipient cells chromosome

Recombinant chromosome

Bacterial plasmids can serve as carriers for gene transfer

Plasmids are small circular DNA molecules that are separate from the bacterial chromosome
F factor is involved in conjugation
When integrated into the chromosome, transfers bacterial genes from donor to recipient When separate, transfers F-factor plasmid

R plasmids transfer genes for antibiotic resistance by conjugation

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F factor (integrated)

Male (donor) cell Origin of F replication Bacterial chromosome F factor starts replication and transfer of chromosome Recipient cell

Only part of the chromosome transfers

Recombination can occur

F factor (plasmid) Male (donor) cell Bacterial chromosome F factor starts replication and transfer

Plasmid completes transfer and circularizes

Cell now male

Plasmids

Sugarphosphate backbone

Nitrogenous base

Phosphate group

Sugar

Nucleotide

DNA
Nitrogenous base

RNA C G A U

C G A T

Sugar DNA Polynucleotide

DeoxyRibose ribose

Growing polypeptide Large ribosomal subunit Amino acid

tRNA Anticodon

mRNA

Codons

Small ribosomal subunit

DNA

is a polymer made from monomers called

(a)

(b)

is performed by enzyme called

(c)

(d) RNA
comes in three kinds called

(e) (f)

molecules are components of

(g)

use amino-acid-bearing molecules called is performed by organelles called

(h)

Protein

one or more polymers made from monomers called

(i)

You should now be able to

1. Compare and contrast the structures of DNA and RNA 2. Describe how DNA replicates 3. Explain how a protein is produced 4. Distinguish between the functions of mRNA, tRNA, and rRNA in translation

5. Determine DNA, RNA, and protein sequences when given any complementary sequence
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You should now be able to

6. Explain the relationship between DNA genotype and the action of proteins in influencing phenotype
Distinguish between the effects of base substitution and insertion or deletion mutations

7.

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You should now be able to

9. Distinguish between lytic and lysogenic viral reproductive cycles and describe how RNA viruses are duplicated within a host cell

10. Explain how an emerging virus can become a threat to human health
11. Identify three methods of transfer for bacterial genes 12. Distinguish between viroids and prions 13. Describe the effects of transferring plasmids from donor to recipient cells
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Hydrogen bond

Partial chemical structure