MICROBIOLOGY

BACTERIAL
BACTERIAL CELL CELL STRUCTURE,
STRUCTURE,
PHYSIOLOGY,
PHYSIOLOGY, METABOLISM
METABOLISM and
and GENETICS
GENETICS
NAME OF LECTURER
NAME OF LECTURE
LECTURER
DATE OF
DATE OF LECTURE

 storage deposits may consist of polysaccharides such

as glycogen, lipid such as poly-B-hydroxybutyrate, or
OUTLINE
polyphosphate. 
I. Cell Structures III. Microbial Growth and
A. Prokaryotic Cell Nutrition
Bacillus and Clostridium
Structure A. Nutritional Requirements
a) Cytoplasmic for Growth  It produces endospore in response to harsh
Structure B. Environmental Factors environmental condition. 
b) Cell Envelope Influencing Growth Endospores
Structure C. Bacterial Growth  Are small, dormant (inactive)
c) Surface Polymers IV. Bacterial Biochemistry and  Asexual spores that develop inside the Bacterial cell
d) Cell Appendages Metabolism (active vegetated cell) as means of survival. 
B. Eukaryotic Cell A. Metabolism
 Are not means of reproduction. 
Structure B. Fermentation and
Thick Protein Coats
a) Cytoplasmic Resporation
Structure C. Pathways from Glucose  makes them highly resistant to;
b) Cell Envelope to Pyruvic Acid o chemical agent
Structure D. Anaerobic Utilization of o temperature change
II. Bacterial Morphology Pyruvic Acid o starvation
A. Microscopic Shapes (Fermentation) o dehydration
a) Cocci (Spherical) E. Anaerobic Utilization of o ultraviolet and gamma radiation
b) Bacilli (Rod- Pyruvate (Oxidation) o desiccation
shaped) F. Carbohydrates Spores
c) Spirochetes Utilization and Lactose  highly refractive bodies in cells. 
(Spiral) Fermentation
B. Common Stains used V. Bacterial Genetics  Are visualized microscopically as unstained area in a cell
for Microscopic A. Anatomy of a DNA and with the use of traditional Bacterial stain
Visualization RNA Molecule           Schaffer-Fulton
a) Gram stain B. Terminology  most commonly used endospore stain 
b) Acid-fast stains C. Genetic Elements and o size
c) Acridine Orange Alterations o shape
d) Calcofluor white D. Mechanisms of Gene o interior location
e) Methylene blue Transfer o one end (terminal)
f) Lactophenol o subterminal, or central.
cotton blue
g) India ink 3) CELL ENVELOPE STRUCTURE
h) Endospore stain
1) Plasma Membrane
1. CELL STRUCTURES  A phospholipid bilayer with embedded proteins that
envelop cytoplasm
A. PROKARYOTIC CELL STRUCTURE  Made of phospholipids and proteins, doesn’t contain
 Bacterial cell is smaller and less compartmentalized than the sterols (except Mycoplasma spp.)
eukaryotic cell.   Act as osmotic barrier
 Various structures are unique to the prokaryotic cell.  Location of electron transport chain

1. CYTOPLASMIC STRUCTURE 2) Cell Wall

 Bacteria do not contain a membrane-bound nucleus  Maintains the shape of the cell
 Their genome consists of single circular chromosome.   Prevents bursting of cell
 Two Major Types of Cell Wall
1) Bacterial Ribosomes 1. Gram-positive
 consisting of RNA and protein.  a) Have very thick protective peptidoglycan
(murein)
 Found free in the cytoplasm attached to the cytoplasmic
 Consist of glycan (polysaccharide) chains
membrane. 
 Alternating N-acetyl-d-glucosamine
 Site of protein biosynthesis.
(NAG) and N-acetyl-d-muramic (NAM)
o they are 70S in size and dissociate into two sub-
acid
units ( 50S and 30S in size)
b) Many antibiotics affective against gram-positive
            S(Svedberg) Units 
organisms (e.g., penicillin)
 refer to sedimentation rates (unit of time) during high-
 Act by preventing synthesis of
speed centrifugation. 
peptidoglycan
 this unit is named THEODOR SVEDBERG the inventor
of ultracentrifuge  Other components that penetrate to exterior of the cell
 S value is Not additive o Teichoic acid (anchored to the peptidoglycan)
o Lipoteichoic acid (anchored to the PM).
2) Cytoplasmic Granules
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 2. Gram-negative
a) Two layers of cell wall  Slime layers
I. Inner peptidoglycan – thinner  Similar to capsules but are more diffuse layers
II. Outer membrane surrounding the cell
o Function:  They are also made of polysaccharides and severe
 Acts as a barrier to hydrophobic either to inhibit phagocytosis or, in some cases, to
compounds and harmful substances aid in adherence to host tissue or synthetic implants.
 Acts as sieve, allows water-soluble
molecules to enter through protein- 5) CELL APPENDAGES
lined channels (porins)
 Provides attachment sites 1) Flagella
o Contains:  Organ of locomotion; seen usually in bacilli
 Proteins  Slender whip like structures → exhibit lashing, forwards
 Phospholipids and rotatory movements
 Lipopolysaccharide (LPS)  Made up of protein called flagellin
a. antigenic O–specific  Parts: basal body, hook, filament
polysaccharide  Classification:
b. core polysaccharide o Monotrichous- single polar flagellum
c. inner lipid A (endotoxin)
o Lophotrichous- a tuft of flagella at one end or at
 responsible for
both ends
producing fever
o Amphitrichous- single flagella at both ends
 shock conditions in
o Peritrichous- flagella all around the bacillus
patients
III. Periplasmic Space o Atrichous- without flagella
o Located between outer and inner
membrane 2) Pili and Fimbriae
o A gel-like matrix; contains nutrient-binding,  Pili (pilus)
degradative and detoxifying enzymes  Known as conjugation pili
o Absent in gram-positive  Non-motile, long, hollow protein tubes
3. Acid-Fast Cell Wall  Connect two bacterial cell
a) Mycobacterium and Nocardia  media DNA exchange
 Gram-positive cell wall  Fimbriae (fimbria)
 Contain waxy layer of glycolipids and fatty  Non-flagellar, sticky, proteinaceous, hair-like
acids (mycolic acid) appendages
 More than 60% (lipid) -> mycolic acid  Adhere some bacteria cells to one another and to
(strong hydrophobic; forms lipids shell and environmental surfaces
affects permeability)
 Stained with: Types:
o Carbolfuchsin  Common pili – thousands of pili around bacteria
o followed by acid-alcohol as a  Sex (F) pili – one or 2 in a bacterium
decolorizer Functions:
4. Absence of Cell Wall  Adhesion- adherence to glycoproteins of GUT;
a) Mycoplasma and Ureaplasma common pili
 Lack of rigidity of the cell wall  Used in transfer of genetic material by the process
 Seen in various shapes microscopically of conjugation (sex pili)
 Contains sterols in their cell membrane  Virulence – ability to infect or damage a host
 Antigenic – can induce antibody production
 Gram-positive and gram-negative cells can lose their cell  Antiphagocytic – prevents action of phagocytes/
walls prevents occurrence of phagocytosis
 Grow as L-forms in media supplemented with serum or
sugar to prevent osmotic rupture of the cell membrane

4) SURFACE POLYMERS

1) Capsule
 Made of polysaccharide polymers; also made of
polypeptides
 Act as virulence factors in helping the pathogen evade
phagocytosis

 Identification of certain bacteria by serologic typing

 Capsule sometimes must be removed to detect the
somatic (cell wall) antigens present underneath them
 Capsule removal is accomplished by boiling a
suspension of the microorganisms.

 Common laboratory stains

 Gram
 India Ink B. EUKARYOTIC CELL STRUCTURE

1. CYTOPLASMIC STRUCTURE

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 1) Nucleus  Found in ciliated epithelial cell of respiratory tract
 Contains DNA in the form of discrete chromosomes  Flagella
(genes)  Longer projections (>150um)
 Covered with basic protein (histones)  Used for locomotion (spermatozoa)

2) Nucleolus  Basal body or kinetosome

 Rounded and refractile body  Small structure
 Located within the nucleus  Located at the base of cilia or flagella
 Site of rRNA synthesis o Microtubule proteins involved in movement
3) Endoplasmic Reticulum (ER) originate
 System of membranes occurs throughout the cytoplasm
o Two forms: 2. BACTERIAL MORPHOLOGY
 Smooth ER
 Don’t have ribosomes 1. MICROSCOPIC SHAPES
 Synthesize phospholipids
 bacteria vary in size from 0. 4 to 2 um. 
 Rough ER
 they occur in three basic shapes. 
 Covered with ribosomes
 Site of protein synthesis
    Three Basic Shapes
4) Golgi apparatus
 Modify and package proteins sent by rough ER
1. Cocci (Spherical) 
5) Eukaryotic ribosomes
 Protein synthesis occurs; attached to rough ER  plural of COCCUS, may occur singly
 80S in size and dissociate into 2 subunits: 60S and 40S  DIPLOCOCCI, in pairs
6) Mitochondria  STREPTOCOCCI, in chains, or;
 Main sites of energy production  STAPHYLOCOCCI, in clusters
 Contain their own DNA and electron transport system
7) Lysosomes 2. Bacilli (Rod-Shaped)
 Contains hydrolytic enzymes; degradation of  plural of BACILLUS
macromolecules and microorganisms  may vary greatly in size and length from very short
8) Peroxisomes COCCOBACILLI to long filamentous rods.
 Contain protective enzymes; breakdown hydrogen  the ends may be square or rounded. Some bacilli are
peroxide curved. 
9) Chloroplasts
 Found in plant cells a) Fusiform
 Sites of photosynthesis (glucose)  is a bacilli with tapered, pointed ends. 
 Site of energy production b) Pleomorphic
 when the species varies in size and shape within a
2. CELL ENVELOPE STRUCTURE pure culture. 
c) Palisading
1) Plasma Membrane  it is called PALISADING when bacilli may occur as
 A phospholipid bilayer with embedded proteins that single rods or in chains or may align themselves
envelop cytoplasm side by side. 
 Regulates transport of macromolecules in and out 3. Spirochetes (Spiral)

 Cholesterol
 Stabilizing effect and helps keep membrane fluid
 Hydrophilic
 Polar heads of phospholipids
 Water loving
 Lie on both the intracellular and extracellular fluids
 Hydrophobic
 Non polar tails
 Water hating
 Avoid water bu lining up in the center of the PM
“tail to tail”
 Proteins
 Perform several important of the membrane;
acts as:
o enzymes
o hormone receptors
o pore channels
o carriers

2) Cell Wall
 Provide rigidity and strength to the exterior of the cell
 Most eukaryotic cells have no cell walls except fungi
 Made of polysaccharides (chitin, mannan, glucan)

3) Motility Organelles
 Cilia
 Short projections (3-10um)
 Numerous, extended from cell surface
 Used for locomotion (protozoa)
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  vary in length and in the number of helical turns (not all a. Zielhl-Neelsen Method  - by heat
helical bacteria are called spirochetes) b. Kinyoun Method - by detergent

 Acidified Alcohol   - is used as a decolorizer

 Methylene Blue - is the counterstain
 Acid-Fast Bacteria  - retain the primary stain and are red
o bacteria that are NOT Acid-Fast are blue. 

                 Two Other Gram-Positive Genera

 Nocardia                  may stain acid fast
 Rhodococcus          by modified method

 Acid-Fast Staining   - is used to identify;

 Saccharomyces
 Yeast
 Coccidian Parasites, such as;
o Cystoisospora belli (formerly known as Isospora belli)
o Cryptosporidium
o and other Coccidia-like bodies 

 Acid-Fast Bacteria   - appear yellow or orange under fluorescent

microscope, making them easier to find. 
 Fluorochrome(i.e.fluorescent) stain   it is also been used to
 Auramine-Rhodamine screen for acid-fast bacteria

3. Acridine Orange
 It is a fluorochrome dye that stains both gram-positive
and gram-negative bacteria, living or dead. 
 It is used to locate bacteria in blood cultures and other
specimens where discerning bacteria might otherwise be
difficult. 

4. Calcofluor White
 it is fluorochrome that binds to chitin in fungal cell walls. 
 Calcofluor White was the original "blueing" used in the
high-volume laundries to whiten yellow appearing white
cotton and other fabrics. 
2. COMMON STAINS USED FOR MICROSCOPIC
VISUALIZATION 5. Methylene Blue
 stains that impart color or fluorescence are needed to  traditionally has been used to stain C. Diptheriae for
visualize bacteria under the microscope.  observation of metachromatic granules.
 It is also used as counterstain in acid-fast staining
1. Gram Stain procedures. 
 most commonly used stain in the clinical microbiology
laboratory 6. Lactophenol Cotton Blue
     is used to stain the cell walls of medically important fungi
Two Main Groups grown in slide culture.
        A. Gram Positive  (blue to purple)
        B. Gram Negative  (pink) 7. India Ink
 Some organisms are gram-variable or do not  is a negative stain used to visualize capsules
stain at all surrounding certain yeasts, such as Cryptococcus spp. 
 The gram stain consists of gentle hat fixing
(methyl alcohol may also be used to fix) of the 8. Endospore Stain 
smear.   to heat-fixed smear, the primary stain, malachite green,
is applied (flooded) and heated to steaming for about 5
minutes. 
Four Sequential Components
1. Crystal Violet - the primary stain, 1 minute Preparation:
2. Iodine - the mordant or fixated, 1 minute  washed for about 30 seconds to remove primary stain
3. Alcohol or an Alcohol Acetone Solution - the  the counterstain Safranin is applied to the smear
decolorizer, quick on and rinse.  o the endospores appear green within pink-appearing
4. Safranin - the counterstain, 30 seconds.  or red-appearing Bacterial cells.

2. Acid-Fast Stains 3. MICROBIAL GROWTH and NUTRITION

 it is used to stain bacteria that have high lipid and wax
content in their cell walls and do not stain well with  All bacteria have three major nutritional needs for growth. 
traditional Bacterial stains. 
 Carbolfuchsin (a red dye) A. Carbon 
o used as the primarily stain  a source of carbon (for making cellular constituents)
 Represents 50 % of dry weight of a bacterium. 
Treated To Allow Penetration Of The Dye B. Nitrogen    

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  a source of nitrogen (for making proteins) o media that are more complex and made of extracts
 Makes up 14% of the dry weight.  of meat or soybeans.
C. ATP                        (i.e., nutrients broth, trypticase soy broth)
 Enriched
 source of energy (for performing cellular functions) o a growth medium that contains added growth factors
such as;
Smaller amounts of molecules that make up an additional
 blood, vitamins, and yeasts (i.e., blood
4% of the weight such as:
agar, chocolate agar). 
 Phosphate for Nucleic acid
 Selective Media 
 Phospholipids of cell membrane o media containing additives that inhibit the growth of
 Sulfur for Protein Synthesis some bacteria but allow others to grow. 
 Differential Media      
 Various metals and ions for enzymatic activity must also
o It is the ingredient in media that allows visualization
be present. 
of metabolic differences between groups or species
of bacteria.
 Important Ions such as:
 Na+
MAC (MacConKey Agar)
 K+ o Can also be a differential medium because it
 Cl- distinguishes between lactose fermenters (pink)
 Ca2+ and non lactose fermenters (clear)
o bacteria vary widely in their ability to use BLOOD AGAR PLATE 
different sources of these molecules.  o can also be non strict, differential because it
distinguishes between hemolytic and non
1. NUTRITIONAL REQUIREMENTS for GROWTH hemolytic organisms. 

 Bacteria are classified into two basic groups according to how  Transport Medium
they meet their nutritional needs.  o It is used when there's a delay between
collection of the specimen and culturing the
 Two Basic Groups specimen is necessary. 
1. Autotrophs (lithotrophs)                    Transport Medium common examples;
 Able to grow simply  Stuart broth
 Using Carbon Dioxide as the sole source of  Amies
carbon, with only water and inorganic salts required  Cary-Blair
in addition.
 Occur in environmental milieus.  2. ENVIRONMENTAL FACTORS INFLUENCING GROWTH
 Obtains energy either;   
o Photosynthetically (Phototrophs)         Three Environmental Factors:
o Oxidation of inorganic compounds 1. pH
(chemolithotrophs)  Most pathogenic bacteria grow best at a neutral pH. 
2. Temperature  
 influences the rate of growth of a Bacterial culture. 

2. Heterotrophs          Category of Microorganism

 requires more complex substance for growth.   according to the optimal temperature for growth
 These bacteria require an organic source of carbon,
such as: I. Psychrophiles
o Glucose  bacteria that grow best at cold
o Obtain energy by oxidizing or fermenting temperatures (optimal growth at 10°Cto
organic substances. 20°C)
II. Mesophiles        
 All bacteria that inhabit the human body fall into the  Bacteria that grow optimally at moderate
heterotrophic group.  temperature. (optimal growth 20°C to
40°C)
I. E. coli and Pseudomonas aeruginosa III. Thermophiles       
 can be used a wide variety of organic compounds as  bacteria that grow best at high
carbon sources and grow on most simple laboratory temperature. (optimal growth to 50°C to
media.  60°C)
II. Haemophilus influenzae and the anaerobes
 are fastidious, requiring additional metabolites such as; 3. Gaseous Composition of the Atmosphere
o vitamins,  Bacteria that grow on humans vary in their
o purine, atmospheric requirements for growth. 
o pyrimidines,
o hemoglobin supplied in the growth medium. A. Obligate Aerobes  - require oxygen for growth. 
B. Aero Tolerant Anaerobes - (previously referred to
III. Chlamydia spp. as Facultative Aerobes)
 cannot be cultured in laboratory media at all and must be o Can survive in the presence of oxygen but do
grown in tissue culture or detected by other means.  not use oxygen in metabolism. 
C. Obligate Anaerobes  - cannot grow in the presence
Types of Growth Media of oxygen
 Minimal Medium   D. Facultative Anaerobes  - can grow either with or
o a laboratory growth medium whose contents are without oxygen. 
simple and completely defined. 
 Nutrients Media 
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 E. Capnophilic  - organisms grow best when the
atmosphere is enriched with extra carbon dioxide
5% to 10%.  carbohydrates.
 Air contains approximately, 21% Oxygen,
1% Carbon dioxide
F. Microaerophilic  - bacteria require a reduced level
of oxygen to grow. (i.e., pathogenic,microaerophiles
are Campylobacter spp. Which requires 5% to 6%
oxygen.  C.
3. BACTERIAL GROWTH
1.  Generation Time  fermentation.
o Bacteria replicate by binary fission, with one cell dividing
into two cells.  o
o The generation time of a bacterium in culture can be 20
minutes for a fast growing bacterium such as E. coli. 
2. Growth Curve     
o If bacteria are in a balanced growth state, with enough
nutrients and no toxic products present, the increase in
Bacterial numbers is proportional to the increase in other
Bacterial properties such as mass, protein content, and
nucleic acid content.
      Four Phases of Growth
 Lag Phase -  during which bacteria are preparing to
divide. 
 Log Phase - during which bacteria numbers
increase logarithmic ally.
 Stationary Phase - in which nutrients are becoming
limited and the numbers of bacteria remain constant
(although viability may decreased)
 Death Phase - when the number of nonviable
Bacterial cells exceeds the number of viable cells. 
3. Determination of Cell Numbers
In the diagnostic laboratory, the number of Bacterial cells
present is determined in one of three ways:
       Three Ways
E.
I. Direct Counting Under The Microscope 
 This method can be used to estimate the number of
bacteria present in specimens. It does not
distinguish between live and dead cells.  o
II. Direct Plate Count 
 This method provides a count of viable cells only. It
is used in determining the Bacterial cell count in
urine cultures.  FERMENTATION
III. Density Measurement
 The density referred to as cloudiness or  turbidity of
a Bacterial broth culture in Log phase can be
correlated to CFU/mL of the culture.  and pH indicator.
 This method is used to prepare a standard inoculum
for antimicrobial susceptibility testing.
4. BACTERIAL BIOCHEMISTRY and METABOLISM
A. METABOLISM
o A type of bacteria uses to break down organic
compounds.
o Depends on the presence and activity of specific
enzymes.
o Can be regulated by either regulating the production of
the enzyme itself or regulating the activity of the
enzyme.
B. FERMENTATION and RESPIRATION
o Bacteria use biochemical pathways to breakdown
o Produces energy by fermentation and oxidation.
o Fermentation, a chemical process by which molecules
like glucose are broken down anaerobically.
o Respiration, an efficient energy generating process in
which molecular oxygen is the final electron acceptor.
o
BIOCHEMICAL PATHWAYS from GLUCOSE to PYRUVIC
ACID
o Glucose, the starting carbohydrate for bacterial
o Pathways are designed to generate pyruvic acid
Has 3 major biochemical pathways bacteria to break
down glucose
 Embden-Meyerhof-Parnas (EMP)
Pentese phosphate pathway
 Entner-Doudoroff pathway
D. ANAEROBIC UTILIZATION of PYRUVIC ACID
(FERMENTATION)
o A key metabolic intermediate
o pathways used by the microbes that inhabit the human
body are:
1. Alcoholic fermentation: the end product is ethanol.
2. Homolactic fermentation: The end product is lactic
acid.
3. Heterolactic fermentation: the end products include
carbon dioxide, alcohols, formic acid, and acetic acid.
4. Propionic acid fermentation: gram positive bacilli is
the end product
5. Mixed acid fermentation: produce lactic, acetic,
succinic, and formic acids as end products.
6. Butanediol fermentation: The end products are
acetoin (acetyl methyl carbinol) and 2,3-butanediol.
AEROBIC UTILIZATION of PYRUVATE (OXIDATION)
o pyruvate is oxidized
o carbon skeletons for biosynthetic reactions are created
Pyruvate donates electron through an electron transport
chain to form ATP.
F. CARBOHYDRATE UTILIZATION and LACTOSE
o Fermentation of the sugar is detected by acid production
o Determination of the microorganism’s ability to ferment
lactose.
o Lactose fermenters or lactose nonfermenters.
5. BACTERIAL GENETICS
A. ANATOMY of a DNA and RNA MOLECULE
 DNA is a double helical chain of nucleotides twisted
together like a spiral staircase
 Has a nucleotide with complex combinations of the
following:
o A phosphate group (PO4)
o A cyclic five-carbon pentose sugar
o A nitrogen-containing base
 A purine consists of adenine and guanine
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  A pyrimidine thymine and cytosine
 A nucleotide is a basic building block of nucleic acid
 Adenine of one chain always pairs with thymine of the
other chain
 Cytosine of one chain pairs with guanine of the other
chain.
 The bases are held together by hydrogen bonds
 The two complementary sugar phosphate strands run
antiparallel 3′ to 5′ and 5′ to 3′.
 RNA is single-stranded and short, contains sugar ribose
 In RNA, the nitrogenous base thymine is replaced by
uracil
 Bacterial genome become a big part in microbiology
 Polymerase Chain in Reaction or (PCR) technique
amplifies specific DNA sequence to detect bacteria
present in the specimen.
B. TERMINOLOGY
 Genotype, a genetic potential of the DNA of an organism
 Protein synthesis is encoded in the bacterial DNA in the
chromosome to each generation of cells
 The general flow of information in a bacteria cell is from
DNA to messenger RNA to the actual protein itself
 Replication is the duplication of chromosomal DNA for
insertion into a daughter cell
 Translation is the actual synthesis of a specific protein
from the mRNA code
 Proteins are polypeptide composed of amino acids
 Codons determines the number of sequences of amino
acid in a polypeptide
 Ribosomes containing rRNA adds amino acids to the
growing polypeptide chain and brought back to ribosome
by tRNA that translate the codons
 tRNA molecules temporarily attach to mRNA using their
complementary anticodon regions
C. GENETICS ELEMENTS and ALTERATION
1. BACTERIAL GENOME
 Consists of a single, closed, circular piece of dsDNA is super
coiled
 Genes are specific DNA sequences that code for the amino
acid sequence in one protein
2. EXTRACHROMOSOMAL ELEMENTS
 Bacteria contain extra information on small circular pieces of
extra chromosomal, dsDNA called plasmids
 Plasmid are not essential for bacterial growth
 Genes that code for antibiotic resistance are often located on
plasmids
 Plasmids are in the cytoplasm of the cell and are self-
replicating just like DNA
 Antibiotics resistance genes are usually located in plasmids
3. MOBILE GENETIC ELEMENTS
 Certain pieces of DNA are mobile also called as jumping
genes
 Simplest mobile pieces of DNA is an insertion sequence (IS)
element
 Each IS element code for only one gene
 Bacterial genomes contain many IS elements
 Transposons are related mobile elements that carry
antibiotics
4. MUTATIONS
 Changes that occur in the DNA code and often result in a
change in the coded protein or in the prevention of its
synthesis
 Mutation can be result of a change in one nucleotide base
 Incomplete, inactive proteins are often the result of mutation
 Mutations also occur as the result of error during DNA
replication
5. MUTATION ALSO OCCUR AS THE RESULT DURING
DNA REPLICATION
D. MECHANISM of GENE TRANSFER
 Can be transferred from one bacterium to another by
o Transformation
o Transduction
o Conjugation
1. TRANSFORMATION
 The uptake and incorporation of naked DNA into a bacterial
cell
 Recombination takes plays by breaking down of DNA and
recombined to produce new combinations
 Cells that can take up naked DNA are referred to as being
competent
 Only a few bacterial species, such as Streptococcus
pneumonia, Neisseria gonorrhoeae, and H. influenza can do
this naturally
2. TRANSDUCTION
 Transfer of bacterial genes by a bacteriophage from one cell to
another
 Step by step on how transduction occurs
o Infection of the bacterial cell by bacteriophage
o The genome of the bacteriophage stably integrates into
the chromosome of the host bacterium and replicates in
concert with it
o When these viruses infect another bacterial cell, they
inject the viral DNA as well as donor DNA into the host
cell
o The bacterial DNA either forms plasmids or gets inserted
into the recipient DNA if it is homologous to the recipient
genome
o In the field of biotechnology, phages are often used to
insert cloned genes into bacteria for analysis
3. CONJUGATION
 The transfer of genetic material from a donor bacterial strain
to a recipient strain, close contact is required
 In the E. coli system, the donor strain (F+) possesses a
fertility factor (F factor) on a plasmid that carries the genes for
conjugative transfer
 The first three letters in the restriction endonuclease name
indicate the bacterial source of the enzyme