Chapter 7: Microbial Genetics

The Structure and Replication of Genomes

The Structure of Nucleic Acids

-DNA and RNA are polymers of repeating nucleotides

-nucleotides are linked through their sugars and phosphates to form backbones.

-the phosphate on the 5’ carbon of the sugar is linked to the 3’ carbon of the next nucleotide. In DNA these strands run antiparallel

The Structure of Prokaryotic Genomes

-Prokaryotic Chromosomes-

-Circular chromosome is concentrated in an area called the nucleoid

-Plasmids are small circular molecules of DNA that replicate independently of the chromosome

-genes on plasmids are generally not essential for normal metabolism, growth, or reproduction.

-the genes on plasmids can confer advantages to the cells that carry them

Gene Function

Regulation of Genetic Expression-

The Nature of Prokaryotic Operons- Operons were first proposed in 1961, the operon consists of

1. promoter:

2. operator:

3. structural genes:

-Inducible operons-

-Repressible operons-

-The Lactose Operon is an example of an Inducible Operon

-it includes a promoter, an operator, and three genes that code for enzymes that are involved in catabolism of lactose.

-The Tryptophan Operon is an example of a Repressible Operon

-it includes a promoter, an operator, and five genes that code for enzymes involved in tryptophan synthesis.

-Research has shown that multiple operons can be controlled by a single repressor, or that transcription rates can vary with the concentrations of corepressors.

Mutations of Genes

Types of Mutations

-point mutations (substitutions, insertions, deletions)

-frameshift mutations

Effects of Mutations

-silent mutation-

-missense mutation-

-nonsense mutation-

Mutagens

-Radiation- can be ionizing or nonionizing.

-Chemical mutagens, three general types

-nucleotide analogs-

-nucleotide altering chemicals-

-frameshift mutagens-

Frequency of Mutation- mutations are rare events.

- one in every 107 genes contains an error.

-mutagens increase mutation rate to one in 104 to 106

DNA Repair- bacterial cells have mechanisms to repair DNA damage which may generate mutations.

-Repair of pyrimidine-dimers: thymine dimers result from exposure to UV light, two adjacent thymines bond together, instead of adenine.

-Base-Excision Repair: a wrong base is inserted during DNA replication

-Mismatch Repair: similar to base-excision, an incorrect base is incorporated into the new DNA strand.

-SOS Response: acts when major damage occurs to DNA that cannot be handled by the regular repair systems. (only in bacteria)

Identifying Mutants, Mutagens, and Carcinogens

-Positive Selection- a mixture of wild type and mutants are grown on a plate that will select against the wild type (Ex. penicillin resistant [mutants] bacteria will grow on a plate with penicillin, wild types will not)

-Negative (indirect) Selection- used for bacteria that have lost the ability to produce a needed metabolite (auxotroph). Follows these steps:

1.

2.

3.

4.

5.

6.

Ames Test for Identifying Carcinogens

-Uses Salmonella that are his-, histidine auxotrophs, follows these steps:

1.

2.

3.

Genetic Recombination and Transfer

Horizontal Gene Transfer among Prokaryotes

-genetic material is passed from a donor cell to a recipient cell of the same generation. (vertical transfer is from parent to offspring)

-recipient and donor can be of the same or difference species or genus.

-the recipient usually incorporates the donor DNA into their chromosome, becoming a recombinant.

-horizontal transfer is rare, occurring in less than 1% of the population, however, there are three types of horizontal gene transfer:

1. Transformation-

2. Transduction-

3. Bacterial Conjugation-

Transposons and Transposition

-transposons are segments of DNA (700-40,000 bp) that move themselves from one location in a DNA molecule to another location in the same or a different molecule.

-transpositions can result in frameshift mutations, rare

-can be simple; containing just the palindromes on either side of the gene for trasposase, or complex; containing two transposons on either side of a non-related gene (kanamycin resistance).

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