Chapter 8  Microbial Genetics

 

 

Genetics – science of heredity  (how genes operate)

 

Chromosomes –

Structures containing DNA,

Carry hereditary information,

Contain genes

Bacteria usually have single circular chromosome

Animals have multiple linear chromosomes

 

Genes – segments of DNA that code for functional products (proteins)

 

Haploid – have one set of chromosomes

Ex:  bacteria

 

Diploid – have two sets of chromosomes

Ex: animals

 

DNA and RNA

Composed of nucleotides

A.  Sugar

B.  Phosphate

C.  Base

Adenine (A)

Guanine (G)

Cytosine (C).

Thymine (T) (DNA only)

Uracil (U) (RNA only)

 

DNA – double stranded

RNA – single stranded

 

DNA exhibits complementary base pairing

A – T

C – G

 

Genotype

Genetic properties (genes)

Potential properties

 

Phenotype

Actual, expressed properties

Can see or measure


The flow of genetic information (DNA replication and gene expression)

 

DNA   transcriptionà   mRNA   translationà    protein

   |

   |  DNA replication

  V

DNA

 

 

DNA replication (short version) (Fig. 8.3)

A.  Enzymes unwind and “unzip” DNA stands

B.  DNA polymerase adds a complementary nucleotide to each nucleotide in the original strands

C.  Two identical DNA molecules are formed

 

 

DNA replication (detailed version)

 

A.  DNA structure

DNA is double stranded

Each strand has 3’ and 5’ ends

The two strands are antiparellel (Fig 8.4)

One strand runs 3’ to 5’

Other strand runs 5’ to 3’

DNA polymerase adds nucleotides to 3’ end only, so DNA replication can only go in the 5’ à 3’ direction

 

B.  Events of DNA replication (Fig 8.6)

 

1.  Enzymes unwind and unzip DNA strands

 

2.  Leading strand synthesized continuously (5’ to 3’) by DNA polymerase

 

3.  Lagging strand synthesized discontinuously

a.  RNA polymerase synthesizes a short RNA primer (provides a 3’ end to build on)

b.  DNA polymerase extends DNA from RNA primer to the last RNA primer that was installed

c.  DNA polymerase replaces previous RNA primer with DNA

d.  DNA ligase does final joining of the fragments

 

 

Gene Expression (mRNA and Protein Synthesis)

 

A.  Transcription (DNA à mRNA)  (Fig. 8.8)

 

1.  RNA polymerase binds to promoter, DNA unwinds

 

2.  RNA synthesized by complementary base pairing


3.  Site of RNA synthesis moves along DNA, transcribed DNA rewinds

 

4.  Transcription reaches terminator

 

5.  RNA and RNA polymerase released, DNA helix reforms

 

B.  Translation (mRNA à protein)

 

1.  Codons

Language of mRNA

Groups of 3 nucleotides

Codons code for amino acids (Fig. 8.9)

Code is degenerate (redundant)

More than one codon can code for the same amino acid

Protects microbe from effects of radiation

 

Type of codons

a.  Sense codons – code for amino acids

b.  Nonsense codons (Stop codons)

UAA, UAG, UGA

Signal the end of protein synthesis

c.  Start codon

AUG

Initiates the synthesis of protein

In addition, codes for Methionine

 

2.  tRNA

Carries correct amino acid to ribosome

Has anticodon on one end that binds to codon of mRNA

Has specific amino acid that is bound onto the other end

 

3.  The process of translation (Fig. 8.10)

(1)  tRNA, ribosomal subunits and mRNA come together

(2)  tRNA with Met bind to start codon on mRNA in P site of ribosome

(3)  tRNA with second amino acid binds to second codon in the A site of ribosome.  Peptide bond forms between amino acids.

(4)  First tRNA is released

(5)  Ribosome move along mRNA until second RNA is in P site.  Third tRNA binds to third codon in A site.

(6)  Process repeated, more amino acids added

(7)  Ribosome reaches stop codon, polypeptide released

(8)  Last tRNA released, ribosome comes apart, polypeptide folds into protein

 

 

Mutation – change in base sequence of DNA (i.e., a change in genotype)

Variation – a change in the appearance of a microbe due to a change in its genotype


A.  Types of mutations

 

1.  Base Substitution (Point mutations) – one base substituted by different base (Fig. 8.16)

 

a.  Silent

No change in amino acid coded for

No effect on finished protein

Ex:    UCA  à   UCG

        serine      serine

 

b.  Missense mutation (Fig. 8.17b)

New codon codes for different amino acid

Ex: sickle-cell anemia due to one missense mutation in hemoglobin gene (Aà T)

 

c.  Nonsense mutation (Fig. 8.17c)

Change in base resulting in a stop codon in middle of mRNA

Translation stops in middle of protein synthesis

 

2.  Frameshift mutations (Fig. 8.17d)

One or more bases deleted or inserted in the DNA

à Translational reading frame shifted

à Coding drastically altered

à Inactive (defective) protein

 

B.  Mutagens – agents that cause mutations

 

1.  Chemical mutagens

 

a.  DNA-modifying agents

Alters bases and makes them bond with incorrect base

Causes point mutations

Ex:  Nitrous acid (Fig. 8.18)

 

b.  Nucleoside analogs

Substitutes in place of base

Analog binds with incorrect base during replication

Causes point mutation

Ex:  2-aminopurine (Fig. 8.19)

 

c.  Frameshift mutagen

Insert between base pairs à cause frameshift mutation

Often potent carcinogens

Ex:      benzpyrene, in smoke and soot

aflatoxin, produced by mold that grows on peanuts


2.  Radiation

 

a.  Ionizing radiation

X-rays and gamma rays

Ionizes cellular constituents

Ions bind with DNA à errors in DNA replication à mutations

 

b.  Non-ionizing radiation

UV

Causes thymine dimers à stops DNA transcription or replication (Fig. 8.20)

Body repairs, but sometimes replaces with incorrect bases à mutation

 

C.  The frequency of mutation

1.  Spontaneous mutations:  1 in 106 replicated genes

2.  Mutagens:  increases 10-1000 times à up to 1 in 103 replicated genes

 

D.  Identifying Chemical Carcinogens à Ames test (Fig. 8.22)

 

1.  Background for Ames test

Prototroph – normal microbe

Auxotroph – prototroph that mutated

à Now can’t synthesize some essential nutrients

Revertant – auxotroph that had a reverse mutation back to prototroph

 

 Prototroph    mutationà  Auxotroph  Reverse mutation  à Revertant

(His + Salm.)             ( His- Salm.)   due to mutagen       (His+ Salm.)

 

2.  Procedure of Ames test

a.  Put auxotroph on Media lacking histidine and containing suspected mutagen

b.  Incubate

c.  Count colonies (any colonies will be revertants)

If count few colonies à due to spontaneous mutations

If count many colonies

à compound is mutagenic

à compound is probably carcinogenic (most carcinogens are also mutagens)

 

 

Genetic Recombination (in bacteria) – the exchange of genes between two DNA molecules

 

A.  Transformation

Genes are transferred from one bacterium to another as “naked” DNA in

solution


Griffith’s genetic transformation experiment (Fig. 8.24)

Be able to describe

 

The mechanism of genetic transformation (Fig. 8.25)

1.  Recipient cell takes up DNA fragments from donor cells

2.  Recombination occurs between donor DNA and recipient DNA

 

B.  Conjugation

Genes are transferred from one bacterium to another by direct contact (often via a sex pilus) (Fig. 8.26)

 

1.  Types of cells involved in conjugation

 

a.  F+  cell

Has F factor on a plasmid

Donor

 

b.  F-  cell

Does not have an F factor

Recipient

 

c.  Hfr cell (High frequency of recombination

Has F factor integrated into chromosome (Fig. 8.27b)

Donor

 

2.  Mechanisms of conjugation (determined by donors)

 

a.  F+ donors (Fig. 8.27a)

(1)  Sex pilus forms

(2)  A copy of plasmid (with F factor) is transferred through sex pilus from F+ to F- cell

(3)  F- cell is converted into an F+ cell

 

b.  Hfr donors (Fig. 8.27c)

(1)  Sex pilus forms

(2)  A copy of chromosome is transferred through sex pilus from Hfr to F- cell (F factor is transferred last)

(3)  Fragile sex pilus usually breaks stopping process before completion

à Part of chromosome transferred, but not F factor

à recipient stays F-

(4)  Recombination occurs between Hfr chromosome fragment and F- chromosome


C.  Transduction

Genes are transferred from one bacterium to another via a bacteriophage or phage (virus that infects bacteria)

 

1.  Generalized transduction – random genes are transferred (Fig. 8.28)

a.  Phage infects donor bacterial cell

b.  Phage DNA and proteins made, bacterial chromosome broken

c.  Occasionally bacterial DNA packaged in viral capsid

d.  Phage infects new host cell (recipient cell)

e.  Recombination occurs between donor and recipient bacterial DNA

 

2.  Specialized transduction – similar process, but only certain bacterial genes are transferred


Chapter 9  Biotechnology and Recombinant DNA

 

 

Biotechnology – the use of microorganisms or cells to make products

 

 

Genetic engineering

= Recombinant DNA technology

= Artificial techniques for exchanging genes between DNA molecules

 

 

Genetic engineering and applications (Fig. 9.1)

 

A.  Gene of interest is cut out of chromosome using restriction enzymes (restriction endonucleases)

 

A.  Gene is inserted into a plasmid

 

B.  Plasmid is taken up by a cell (usually bacterium)

 

C.  Cells cloned (copied) during log phase of bacterial growth

 

D.  Products harvested

 

1.  Genes harvested

Copies of genes are inserted into other organisms to give desirable properties

Ex:  gene for pest resistance is inserted into plants

 

2.   Protein harvested

Altered cells make a protein product

Ex:  Various enzymes, pharmaceuticals

 

 

Pharmaceutical Products of Genetic Engineering (Table 9.1)

Know at least three products


Chapter 12  The Eukaryotes:  Fungi, Algae, Protozoa, and Helminths

 

Mycology  - study of fungi

 

 

Fungi

Eukaryotic

Cell wall of chitin

Chemoheterotrophs

Most are saprophytes (saprotrophs) à feed on dead organic matter

 

A.  Three morphological groups of fungi

 

1.  Molds

Consist of hyphae à long filaments of cells joined together

Mass of hyphae à mycelium

Almost all are aerobic

 

a.  Types of hyphae based on septa (cross walls) (Fig 12.1)

(1)  Septate hyphae – contain septa

(2)  Coenocytic hyphae – no septa, long continuous cells with many nuclei

 

b.  Types of hyphae based on function (Fig 12.2)

(1)  Vegetative hyphae – obtain nutrients

(2)  Reproductive or aerial hyphae – reproduction, bears spores

 

2.  Yeasts

Nonfilamentous, unicellular

Most are facultative anaerobes

 

Reproduce asexually by

a.  budding - dividing unevenly (Fig. 12.3)

Ex:  Saccharomyces (Brewer’s yeast)

b.  fission - dividing evenly

 

3.  Dimorphic fungi

Can grow either as a mold or a yeast (Fig. 12.4)

 

B.  Life cycle of Fungi

 

1.  Asexual Reproduction

Produces asexual spores formed by the hyphae of one organism (Fig. 12.5)

 

a.   Conidiospore 

Not enclosed in a sac

Produced in chains at end of aerial hyphae

 


b.  Arthrospore

Not enclosed in a sac

Formed by fragmentation of hyphae

 

c.  Blastoconidia

Not enclosed in a sac

Formed by buds coming off the parent cell

 

d.  Chlamydospore

Thick-walled spore formed within hyphae

 

e.  Sporangiospore

Enclosed in a sac at end of aerial hyphae

 

2.  Sexual reproduction

Produces sexual spores

 

a.  Process

(1)  Plasmogamy

Haploid nucleus of a donor cell (+) penetrates the cytoplasm of a recipient cell (-)

(2)  Karyogamy

The (+) and (-) nuclei fuse to form a diploid zygote nucleus

(3)  Meiosis

Diploid nucleus à haploid nuclei (sexual spores)

 

b.  Types of sexual spores

(1)  Zygospores

(2)  Ascospores

(3)  Basidiospores

 

C.  Medically important Phyla of Fungi

Phyla determined by sexual spores produced

 

1.  Zygomycota (Fig. 12.6)

Produce zygospores

Saprophytic molds (obtains nutrients from dead organic matter)

Coenocytic hyphae

Ex:  Rhizopus nigricans (common black bread mold)

 

2.  Ascomycota (Fig. 12.7)

Produce ascospores enclosed in a saclike ascus

Molds with septate hyphae and some yeasts

Ex:  Histoplasma capsulatum (causes Histoplasmosis)

 

3.  Basidiomycota (Fig. 12.8)

Produce basidiospores on base pedestal called a basidium

Ex:  “mushrooms”


4.  Deuteromycota

“holding category” for fungi whose sexual cycle has not been observed yet

 

D.  Fungal diseases à mycoses

 

1.  Systemic mycoses

Infections deep within body à Affect tissues and organs

Transmission by inhalation of spores

Ex:  Histoplasmosis

 

2.  Subcutaneous mycoses

Infections beneath skin

Transmission by spores entering puncture wound

Ex:  Sporotrichosis

 

3.  Cutaneous mycoses

Infection of the skin

Transmission by direct contact

Ex:  Tinea capitis (ringworm) (Fig 21.16a)

 

4.  Superficial mycoses

Infection of hair and dead layers of epidermis

Transmission by direct contact

Ex:  Tinea versicolor

 

 

Algae

Unicellular or multicellular

Eukaryotic

Photoautotrophs

Examples:

A.  Brown algae (kelp) (Fig. 12.11b)

B.  Green algae (green pond scum)

C.  Dinoflagellates (Fig. 12.14)

à some produce neurotoxin

à mussels and clams concentrate

à paralytic shellfish poisoning (PSP)

 

 

Protozoa

Unicellular

Eukaryotic

Chemoheterotrophic

 

A.  Many can have two forms

1.  Trophozoite – feeding and growing stage

2.  Cyst – have protective capsule that forms during adverse conditions


B.  Medically Important Phyla of Protozoa

Classified according to type of motility

 

1.  Archaezoa (old name à Mastigophora)

Lack mitochondria

Move by flagella

Ex:  Trichomonas vaginalis (Fig. 12.17b) causes STD

Ex:  Giardia lamblia (Fig. 12.17c)

 

2.  Rhizopoda  (old name à Sarcodina)

Move by pseudopodia

Ex:  Amoeba proteus (Fig 12.18a)

 

3.  Apicomplexa (old name à Sporozoa)

Not motile

Obligate intracellular parasites

Ex:  Plasmodium vivax – causes malaria

Life Cycle (Fig. 12.19)

a.  Infected mosquito bites human and transmits P. vivax

b.  Asexual reproduction in liver and red blood cells

c.  Uninfected mosquito bites infected human à mosquito infected

d.  Sexual reproduction in mosquito

 

Some Protozoa require 2 types of hosts

a.  Definitive host – harbors sexual reproducing stage(in this case, the mosquito)

b. Intermediate host – harbors asexually reproducing stage(in this case, the human)

 

4.  Ciliophora

Move by cilia

Examples:

Paramecium (Fig. 12.20a)

Balantidium coli

Only ciliate that is a human parasite

Causes Ciliary dysentery

 

5.  Euglenozoa (old name à Mastigophora)

Move by flagella

Grouped together because of common genetic sequences

Examples:

Euglena (Fig. 12.21)

Trypanosoma cruzi à Chaga’s disease


Helminths

Parasitic worms

Multicellular

Eukaryotic

Arrangements of reproductive organs

à Dioecious – separate males and females

à Monoecious – one individual has both male and female reproductive organs

 

A.  Platyhelminths – flatworms

Most are monecious

 

1.  Trematodes – flukes

Flat, leaf-shaped bodies

Have ventral and oral suckers

Ex:  Lung fluke (Fig. 12.26)

Occurs throughout world

 

Life cycle

a.  Adult fluke lives in bronchioles of human à Reproduces sexually (definitive host)

b.  Eggs à swallowed sputum à feces à water

c.  Egg hatches à snail à reproduces asexually (int. host)

d.  Leave snail à encyst in crayfish (intermediate host)

e.  Undercooked crayfish eaten by human

 f.  Bores out intestine and into lung

 

2.  Cestodes – tapeworms (Fig. 12.27)

Intestinal parasites

Lack a digestive system à absorb food through cuticle

Has scolex (head) with suckers and hooks

Body consists of segments called proglottids

 

Ex:  Taenia saginata “beef tapeworm” 

Life Cycle

a.  Adults live in human intestines (6 m long) (Defin. host)

b.  Mature proglottids and eggs shed in feces

c.  Cattle eat eggs (intermediate host)

d.  Eggs hatch, larvae bore through intestines to muscle

e.  Human eats beef

f.  Scolex anchors in small intestine

 

B.  Nematodes – roundworms

Cylindrical and tapered at each end

Complete digestive system (mouth, intestines, anus)

Most dioecious


1.  Pinworm (Fig. 12.29)

1 cm long

No damage to host

Life cycle: 

a.  Adults live in human large intestine

b.  Lay eggs around anus

c.  Eggs ingested by contaminated hands or clothing

 

2.  Ascaris lumbricoides

30 cm long

Life cycle:

a.  Adults live in human small intestines

b.  Eggs in feces (can be viable in soil for 10 years)

c.  Eggs ingested

d.  Eggs hatch in small intestine

e.  Larvae burrow out of intestines, enter blood

f.  Carried to lungs and grow

g.  Coughed up, swallowed, and return to small intestine

h.  Mature into adults

 

3.  Hookworms (Fig. 12.30)

Life cycle:

a.  Adults live in human small intestine

b.  Eggs in feces

c.  Larvae hatch in soil

d.  Larvae penetrates hosts skin (barefoot)

e.  Larvae enters blood, carried to lungs

f.  Coughed up, swallowed, and carried to small intestine

g.  Mature into adults

 

4.  Trichinella spiralis

Causes trichinosis

Life cycle:

a.  Larvae encysted in muscle of pigs or wild game

b.  Other animal (or human) eats undercooked infected meat

c.  Mature and reproduce in human small intestine

d.  Larvae migrate and encyst in tissue

 

 

Arthropods as Vectors

Vector = Arthropods that carry pathogenic microorganisms

 

Arthropods

 

A.  Arachnida

8 legs

Spiders, mites, ticks

Ex:  Ixodes (tick) carries microorganism that causes Lyme disease (Fig. 12.32)


B.  Insecta

6 legs

Bees, flies, mosquitoes, lice

Ex:  Anopheles (mosquito) carries microorganism that causes malaria (Fig. 12.31)