Gene Transfer,Genetic Engineering,and Genomics

1. Genetic recombination.

2. Genetic engineering.
3. Microbial genomics.

A. Genetic Terminology

Genotype
The genetic compliment of an organism Types of genotypic changes Mutation Conjugation Transduction Transformation

Phenotype
The genetic expression of an organism Types of phenotypic expressions Morphology Cultural Physiological

B. The Bacterial Chromosome

Introduction
DNA is arranged as a single molecule with no histones present, and with no dominance or recessiveness in the genes. Bacterial chromosome is located in the nucleoid. In E. coli there are 4000 genes spread over 1.5mm of DNA in less than 1 micrometer of space

Loop domain structure allows for compaction of DNA

Replication of the chromosome

DNA polymerase The semiconservative method Replication of a closed loop chromosome Okazaki fragments Rolling circle method

Plasmids
Fragments of DNA in the cytoplasm R Factors - confer drug resistance Bacteriocins -proteins toxic to other bacteria and human cell Many plasmids are found in GramNegative bacteria

C. Bacterial Mutation

Permanent alteration in the DNA

Example: nonpathogenic Yersinia pestis have genes that cause them to remain in mid gut, pathogenic Y. pestis do not have these genes

Types of mutations
Spontaneous Induced

Spontaneous mutations
Occurs every 106 to 1010 replications 1 mutation in every billion bacteria Example: Neisseria gonorrhoeae penicillin resistance original mutation was spontaneous Example: Salmonella strains antibiotic resistance

Induced mutations
Chemical or Physical agents enhance mutation rate Mutagens Ultraviolet lightmechanism of action Chemicals Chromosomal changes

Mutation Type Point (substitution)

(leu) (ser) (arg)

Normal AAT AGT GCC

(leu) (cyst) (arg)

Mutant AAT TGT GCC

Mutation Type Frameshift (deletion)

(leu) (ser) (arg)

Normal AAT AGT GCC

(leu) (val) (pro)

Mutant AAT AGT GCC A

Mutation Type Frameshift (insertion)

(leu) (ser) (arg)

Normal AAT AGT GCC

(leu) (glut) (cyst)

Mutant AAT CAGT GCC

Repair Mechanisms

DNA repair enzymes Many enzymes Constantly checking for errors Repair mechanisms Mismatch repair proofreads Damage repair Excision repair Dimer repair (UV light)

Mismatch Repair

Excision Repair

Transposable genetic elements

Insertion sequences Small DNA segments Provide no genetic information Located at several places on the chromosome Transposons Larger than Insertion sequences Provide information for protein synthesis

Sections A & B repeating but reversed palindrome

Ames Test

C. Bacterial Recombination

Transformation
Description Griffith's experiments Modern interpretation: Avery,McLeod & McCarty Mechanism Competence

Conjugation

Male and female cells Role of F factors (plasmids) High frequency of recombination strains Mechanism of Hfr conjugation Sexduction

Simple Conjugation

Hfr Conjugation

Transduction
Description Role of the bacteriophage The lytic cycle Lysogeny Generalized transduction

Lysogeny

F. Control of protein synthesis

 1. Mechanism proposed by Jacob and Monod 2. The operon theory

3. Repressor-inductor model

D. Genetic Engineering

Genetic Engineering
Genetic Engineering Was Born from Genetic Recombination Genetic engineering involves changing the genetic material in an organism to alter its traits or products
A recombinant DNA molecule contains DNA fragments spliced together from 2 or more organisms

History of Genetic Engineering

Discovery of endonucleases Plasmids and sticky ends

Modern applications

Pharmaceutical production Insulin, interferon, hormones, vaccines etc. Genetically engineered plants Animal gene alterations Gene probes DNA fingerprinting The human genome initiative

E. Genomics

Microbial Genomes Have Been Sequenced Hundreds of microbial genomes have been sequenced since the first in 1995
Many of which are pathogens

Segments of the Human Genome May Have Microbial Ancestors As many as 200 of the 35,000 human genes are essentially identical to those of Bacteria
They were passed down from early ancestors of humans

Microbial Genomics Will Advance Our Understanding of the Microbial World Knowing genomes of bacteria that cause foodborne diseases can help us:
develop detection methods make food safer

It can help us identify microbes that cannot be cultured in the lab Environmental genomics helps us understand how microbial communities function

Microbial Genomics Will Advance Our Understanding of the Microbial World Environmental genomics can help develop bioremediation techniques

Genomics can help develop detection methods for potential bioweapons organisms and other agents of warfare

Comparative Genomics Brings a New Perspective to Defining Infectious Diseases Studying Evolution

Types of Genomics Functional genomics attempts to discover: the function of proteins coded for in a genome how the genes interact, allowing the microbe to grow and reproduce Comparative genomics compares the DNA sequence of one microbe to another similar or dissimilar organism