Microbial Genetics

Replica Plating
negative/indirect selection
selects a cell which cannot perform a specific function

 Selective and differential

medium Used for growing nonfastidious G(-) rods

Presence of bile salts

and crystal violet

 Usually contain the

minimum nutrients possible for colony growth, generally without the presence of amino acids

2 Uses: 1. to grow "wild type" microorganisms 2. to select for or against recombinants

Three components: 1. carbon source (e.g.: sugar) 2. various salts 3. water

 Lost

capability to synthesize an amino acid

 Prototroph- The ability to synthesize all needed

amino acid Wild type Antibiotic resistant- Gained resistance to an antibiotic

 Are agents in the environment, such as certain

chemicals and radiation, that directly or indirectly bring about mutations Almost any agent that can chemically or physically react with DNA can potentially cause mutations.

 Nitrous Acid

 Still, other chemical mutagens cause small deletions

or insertions FRAMESHIFTS Ex: benzopyrene (smoke and soot) aflatoxin (Aspergillus flavus)- mold in peanut and grain acridine dyes used experimentally against herpes virus infections

 X rays and gamma rays potent mutagens because of

their ability to ionize atoms and molecules.

Electrons pop out of usual shells electrons

bombard other molecules and cause more damage ions and free radicals may combine with bases in DNA errors in replication and repair that produce mutations
Breakage of covalent bonds in the sugar-phosphate

backbone physical breaks in chromosomes.

 Ultraviolet (UV) light- a nonionizing component of

ordinary sunlight Most mutagenic component of UV light (wavelength 260 nm) is screened out by the ozone layer of the atmosphere Formation of harmful covalent bonds between certain bases THYMINE DIMERS May cause serious damage or even death because it cannot properly transcribe or replicate such DNA

 Photolyases- also known as light-repair enzymes,

use visible light energy to separate the dimer back to the original two thymines Nuc1eotide excision repair and action of methylases (Hamilton Smith, 1970)
Enzymes cut out incorrect base and fill in the gap with

newly synthesized DNA that is complementary to the correct strand Methylases add methyl group to selected bases soon after DNA is made. A repair endonuclease then cuts the nonmethylated strand.

Two ways in which DNA can become mutated: Inherited- if a parent has a mutation in his or her DNA, then the mutation is passed on to his or her children. Acquired-happens when environmental agents damage DNA, or when mistakes occur when a cell copies its DNA prior to cell division.

 Base substitution/point mutation-single base at one

point in the DNA sequence is replaced with a different base May cause amino acid substitution in the synthesized protein missense mutation or creation of a nonsense (stop) codon in the middle of an mRNA moleculenonsense mutation

 Frameshift mutations-where one or a few

nucleotide pairs are deleted or inserted in the DNA

Can shift the translational reading frame

Spontaneous mutations may occur due to occasional mistakes made during DNA replication. - apparently occur in the absence of any mutationcausing agents