MICROBIAL GENETICS

PART 1
 Genetic Material
 Central Dogma
 Genes Expression
 TERMINOLOGI

 Genetika : the study of what genes are, how they

carry information, how their information is
expressed, and how they are replicated
and passed to subsequent generations or
other organisms

 Genes : Segments of DNA (RNA in some viruses)

that code for functional products. Gene is
basic unit of heredity
 DNA : Makromolekul yang tersusun dari unit –unit yang
disebut sebagai nukleotida.

Nukleotida : Each nucleotide has three parts

 Basa nitrogen : Adenine, Cytosine, Guanine, Thymine
 Deoksiribosa : gula pentosa yang kehilangan satu
 Fosfat

 Chromosomes : Typically a circular (in Prokaryotes) or

linear (in Eukaryotes) thread like molecule of DNA.
Chromosomes contain the genes.
ALL GENES = DNA ?
or
ALL DNA = GENES ?
 2 strands twisted into a helix (double helix)
 Sugar - phosphate backbone
 Nitrogenous bases form steps in ladder
 constancy of base pairing
 A binds to T with 2 hydrogen bonds
 G binds to C with 3 hydrogen bonds
 Antiparallel strands 3’ to 5’ and 5’ to 3’
 One strand is the complement of the other
 Order of bases constitutes the DNA code
• rRNA • Combines with certain proteins to make
(Ribosomal RNA) ribosomes
• Provides a site for protein synthesis
 Copy of DNA information for synthesis of
 mRNA proteins
(Messenger RNA) Corresponds to one or more genes in DNA
 Have codons (triplet bases)
 One or more ribosome can attach at a time
* tRNA * Pick up amino acids in cytoplasm and transfer
them to mRNA
(Transfer RNA)
* Cloverleaf shape with attachment site for a
specific amino acid
* Has anticodon (triplet bases) complementary to
the codon in mRNA
 Ekstra-kromosomal molekul dari DNA yang dapat
melakukan replikasi sendiri.

 Membawa gen-gen yang tidak memiliki peran inti

dalam keberlangsungan hidup sel.

 Berukuran 1-5% dari ukuran kromosom bakteri.

 Ditemukan terutama pada bakteri

 TYPES OF PLASMIDS

1. Conjugative Plasmids
A prokaryotic plasmid that carries genes for sex
pili and for transfer of plasmid to another cell e.g.,
F factor plasmid

2. Dissimilation Plasmids
May code for enzymes that trigger the catabolism
of certain unusual sugars and hydrocarbon e.g.,
some Pseudomonas can use hydrocarbons of
petroleum as their primary carbon and energy
source
 PLASMIDS

3.Plasmids carrying genes for toxins or bacteriocins

These enhance the pathogenicity of a bacterium
e.g., E. coli that cause infant diarrhea and
traveler's diarrhea carry this plasmid for toxin
production and attachment to intestinal wall
(without this plasmid, it is nonpathogenic).
Bacteriocins are proteins toxic for other bacteria

4. Resistance factor (R factor) plasmids

Carry genes that cause resistance to antibiotics or
cellular toxins.
 C
E
DNA N
T
Replication R
A
DNA
L
Transcription
D
RNA O
G
Translation M
A
PROTEIN
 It takes place before every cell division so that each
daughter cell get a copy of the complete genetic material

 Two strands of the double helix separate to form a

Replication Fork (the point at which replication occurs)

 Each strand then serves as a template for the synthesis of

a new strand following the rules of complementary base
pairing
 The process of DNA replication is Semiconservative
as each double stranded DNA molecule has one
original mother strand and one newly synthesized
strand

 DNA Polymerase enzyme can add new nucleotides at

the 3`end of the newly synthesized strand only; so the
DNA is synthesized in one direction designated as

 5` 3`only
R
E
P
L
I
C
A
T
I
O
N
18
 Both strands of DNA are antiparallel (oriented in opposite direction
relative to each other)
3` 5` (Template)

5` 3` (leading strand)

 At the replication fork, leading strand is synthesized continuously as

DNA Polymerase enzyme moves towards the replication fork

 Lagging strand synthesis is discontinuous as it is made in fragments

(called Okazaki fragments) and the DNA Polymerase enzymes move in
opposite direction of the growth of replication fork
 To add a new nucleotide to the growing strand of DNA, a
nucleoside triphosphate bonds to the sugar and loses
two of its phosphates. Hydrolysis of the phosphate
bonds provide energy for the reaction.

 In some bacteria like E. coli, two replication forks move

in opposite direction, so the replication is Bidirectional.
Two forks ultimately meet each other and the replication
is complete
 Helicase enzyme opens the double helix up, so a
replication fork is made (some other proteins are also
involved)
 RNA Polymerase makes a short piece of RNA (called
RNA Primer) on the DNA template
 DNA Polymerase III then adds nucleotides at the 3`
end of the RNA Primer & make the daughter strand. It
also does the proof reading to avoid mistakes in base
pairing
 DNA Polymerase I removes RNA Primer and replaces it
with DNA nucleotides but cannot connect the two newly
made segments together
 DNA Ligase joins the newly made DNA fragments
 DNA Gyrase creates twists or supercoils of DNA
 GENES EXPRESSION

 Genotype :
The entire DNA or collection of genes. It is the entire
genetic material of an organism. It represents the
POTENTIAL properties

 Phenotype :
It refers to the actual, EXPRESSED properties of an
organism. It is an organism’s collection of proteins
 It is the synthesis of complementary strand of RNA from
a specific gene (DNA template). Only one of the two
strands of DNA serve as template
 There are three kinds of RNAs in bacteria
 mRNA, tRNA, rRNA
 Transcription requires
 RNA polymerase
 A supply of RNA nucleotides
 RNA Polymerase binds only to the Promoter Site on the
DNA
 RNA is synthesized in the 5` 3` direction too
 RNA synthesis terminates when RNA Polymerase
reaches a site on the DNA called the Terminator Site
 Protein synthesis is called translation because it involves
decoding the “language” of nucleic acids and converting that
information into the “language” of proteins.
 Language of mRNA is in the form of Codon (groups of three
nucleotides e.g., UCC AUC)
 Each codon codes for a particular amino acid (code is called
the genetic code)
 Initiator codon is AUG
 Terminator (stop or nonsense) codons; UAA, UGA, UAG
 Translation machinery includes :
 mRNA
 tRNA (which has anticodon) carrying amino acids
 Ribosome (have rRNA and specific proteins)
** Ribosomes moves along the mRNA in the 5` 3` direction
Genetic code
 STEPS INVOLVED IN TRANSLATION

Necessary components assemble. Initiator codon (AUG) is the

point where translation begins
The first tRNA binds to the start codon, bringing the amino
acid methionine. Second tRNA arrives on second codon (it
must have complementary anticodon to the second codon)
The first amino acid is transferred by the ribosome to the
Second tRNA (peptide bond is formed)
Empty tRNA leaves. Ribosome then moves along the mRNA
to the next codon toward 3` direction. New tRNA arrives with
next amino acid
As the proper amino acids are brought into line one by one,
peptide bonds are formed between them, and a polypeptide
chain is produced. Translation end at one of the stop codons
STOP CODON (UAA, UAG, or UGA) ON m RNA SEPARATES THE
SYNTHETIC MACHINERY
PROKARYOTIC TRANSLATION
mRNA Processing Before Translation In Eukaryotes
 EUKARYOTE PROKARYOTE
Chromosome > 1, Linear 1, Circular
mRNA Monocistronic Polycistronic
DNA • Contains large regions of • Rarely contains any
repetitive DNA "extra" DNA
• Much of eukaryotic DNA • Often more than 95% of
does not code for proteins the genome codes for
proteins

Genes Split into exons and introns Almost never split

Transcription & Do not occur simultaneously Can take place
Translation simultaneously
Post- √ -
transcriptional
process
Start signal for Methionine Formyl-methionine
protein synthesis
REPLICATION → DNA AMPLIFYING

POLYMERASE CHAIN REACTION

(PCR)
PCR Animation
Please click here.

Process

Denature

Anneal Primer

Replicate
DNA

1st cycle 2nd cycle 3rd cycle

 MICROBIAL GENETIC PART 2

• REGULATION OF GENE EXPRESSION

• MUTATION
• GENE TRANSFER