MICROBIAL GENETICS

Two types of organism:

Eukaryotes; cells contain membrane-bound
compartments, including a nucleus and organelles.
Eukaryotes include: animals, plants, fungi, and
protozoa.
Prokaryotes: Lack internal compartments extensively.
Divided into two groups: bacteria and archaea.

Genome:
The entire genetic complement of a living organism.

Gene:
A DNA segment containing biological information and
hence coding for an RNA and / or polypeptide molecule.

Eukaryotes genomes;
Divided into two or more linear DNA molecules.
Each contained in a different chromosome.
Possess smaller, usually circular, mitochondrial
genomes.
Third genome located in the chloroplast (in plant and
other photosynthetic organism)
10 Mb 100,000 Mb in length.
Higher eukaryotes need larger genome to
accommodate the extra genes.
Correlation between genome size and complexity (???)
--- C-value paradox.
Space is saved in the gnomes of less complex
organism because the genes are more closely packed
together.

Prokaryotes genomes:
Whole prokaryotes genomes are smaller than
eukaryotes.
Most if not all -- are contained in a single DNA
molecules.
The molecule is circular.
Have second circular or linear genome, called
PLASMID.
Have fewer genes
More compact genome organization, more gene but
less space.
There is NO INTRONS (some exception is in
archaea.)
Infrequency of repetitive sequences.

Bacterial DNA structure:

Consists of two chains of chemical units called
nucleotides.
May contain 10 40 million nucleotides.
A nucleotides consists of a base (purine and
pyrimidine)
Purine : adenine (A) and guanine (G)
Pyrimidine: Thymine (T) and cytosine (C)
DNA contains a specific sequence of purine and
pyrimidine base.
A binds to T
G binds to C

upstream

exon intron exon

Initiation codon

downstream

termination codon

Circular shape of microbial DNA

DNA and Genetics Code:

The sequence of codons on the mRNA contain
the necessary information to :
Initiate polypeptide synthesis.
Designate the sequence of amino acids in the
polypeptide.
Terminate the synthesis of the polypeptide.
Release the completed polypeptide.

Genetics codes

DNA Replication:
The transmission of genetic material from parent to
progeny requires an exact duplication of the original
DNA strands.
Major errors in this process leading to prevent normal
cell growth and development.
Each strand of the parent DNA serves as template
for the formation of a complementary strand.

DNA transcription:
The process in which one strand of DNA serves as a
template for the synthesis a single-stranded
messenger RNA (mRNA).
It is the information on the RNA that can be
translated into usable information by the cell.
DNA

DNA

mRNA
rRNA
tRNA

Protein

Translation of the genetic code: Protein

synthesis
mRNA is translated into polypeptides in a process
that take place on the ribosome.
Ribosomes are small structural components in the
cell that are composed of rRNA and a variety of
proteins.
Before protein synthesis begins, each amino acids
was brought to the ribosome by tRNA.
The protein synthesis of the polypeptide takes place
by the linear movement of ribosomes along the
mRNA.

Mutation

The alterations in the base sequence of the hereditary

material (DNA)

Spontaneous mutations

Occur in the absence of environment intervention.

It is occur in case of:
Purine or pyrimidine bases temporarily exist in different
electrochemical forms -------- mispairing.
Insertion sequence.
Missmatch during replication

Induced mutations
Physical agents:
Ultraviolet light (UV)
X-rays
Gamma-rays
Chemical agents

Spontaneous mutation

A IS A

IS

IS

Movement of insertion sequence (IS)

IS element replicates and inserts a copy within the A gene

Mismatch during replication

Mutagenic agent caused mutation

How mutations affect the genetics code ?

Silent mutation (has no effect)

Missense mutation (change to a different amino acids)
Nonsense mutation (change to a stop codon)
Frameshift mutation.

Open reading frame (ORF)

How to read the codons sequences.

DOGCANRUN
DOG-CAN-RUN
D-OGS-ANR-UN
DO-GCA-NRU-N

(Frame I)
(Frame II)
(Frame III)

DOGCANRUN
DOG-CAR-UN

(Frameshift)

The effect of mutations on microorganism:

Auxotrophs
Microorganism that will only grow when provided with a
nutrient that is not produced by it self. Prototrophs are the
wild type microorganism.
Conditional-lethal mutants : permissive condition,
restrictive condition, temperature-sensitive.
Inhibitor-resistant mutants: able to resist the toxic effects
of an antibiotic or other type of inhibitor.
Regulatory mutants: has mutation in the promoter or other
regulatory sequence. EX: constitutive mutants of E. Coli
expressed genes all the times.

Mechanism of gene transfer in prokaryote

Conjugation:
Only one strand of DNA is transferred. The recipient
completes the structure of double stranded DNA.
Transduction:
Donor DNA was carried in a phage coat and is
transferred into the recipient by the mechanism used
for phage infection.
Transformation:
Direct up take of the donor DNA by the recipient cell,
may be nature or forced.

Conjugation

Transposable elements :

Insertion sequences (IS)

Segment of DNA
Approx. 1000 bp
Contain genes need for transposition only
Encode enzyme for site-specific recombination.
Has certain sequence in its termini.
Called replicative recombination
Transposons (Tn element)
10 fold of IS in length.
Has more complex enzyme/proteins
Transposable proghages
Certain phage, ex: Mu

DNA viruses

RNA viruses

Baltimore Classification of Viruses

Group

Genome

dsDNA

ssDNA

dsRNA

+ve ssRNA

dsRNA

-ve ssRNA

dsRNA

ssRNA

Replication
dsDNA

Example
mRNA

ssDNA

dsDNA
dsRNA

Herpes simplex
virus

mRNA
mRNA

Parvovirus
Reovirus

+ve ssRNA [Acts as mRNA] Enterovirus

-ve ssRNA

mRNA

Influenza A
virus

dsDNA

mRNA

Retrovirus
(e.g. HIV)

intact dsDNA

mRNA

Hepatitis B
virus

ssRNA

Nicked dsDNA nicked dsDNA

RNA

Positive Strand RNA Virus Genome

Negative strand RNA Virus Genome

Virus enzyme
Segmen
RNA (-)

Segmen
RNA (+)

Segmen
RNA (-)

Virus baru

mRNA
Strand (+)

Protein