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Define genetics, genome, chromosome, gene, genetic code, genotype, phenotype, and genomics. Describe the process of DNA replication. Describe protein synthesis, including transcription, RNA processing, and translation. Classify mutations by type, and describe how mutations are prevented and repaired. Define mutagen. Describe two ways mutations can be repaired. Outline methods of direct and indirect selection of mutants. Identify the purpose and outline the procedure for the Ames test. Compare the mechanisms of genetic recombination in bacteria. Differentiate between horizontal and vertical gene transfer. Describe the functions of plasmids and transposons.
2004 by Jones and Bartlett Publishers
Terminology
Genetics
Complementary but antiparallel
Genome
Gene Chromosome Base pairs Genetic code
Genomics
Genotype
Phenotype
DNA
E. coli
Fig 8.1
Copyright 2006 Pearson Education, Inc., publishing as Benjamin Cummings
Figure 8.1a
Figure 8.1b
DNA Replication
DNA polymerase initiated by RNA primer
bidirectional
origin of replication leading strand: continuous DNA synthesis lagging strand: discontinuous DNA synthesis Okazaki fragments semiconservative
2
Replication fork
Replication in 5' 3' direction Fig 5.8
Fig 8.6
Protein Synthesis
Genetic code: universal and degenerate (or redundant)
Fig 8.8
Transcription
Fig 8.7
Translation
Fig 8.9
Transcription
RNA polymerase binds to promotor sequence proceeds in 5' 3' direction stops when it reaches terminator sequence
Fig 8.7
Copyright 2006 Pearson Education, Inc., publishing as Benjamin Cummings
More Details on Translation Nucleotide sequence of mRNA is translated into amino acid sequence of protein using three letter words = codons Translation of mRNA begins at the start codon: AUG Translation ends at a stop codon: UAA, UAG, UGA Requires various accessory molecules and 3 major components: ? In Prokaryotes: Simultaneous transcription and translation Polyribosomes
Copyright 2006 Pearson Education, Inc., publishing as Benjamin Cummings
Mutations
Change in genetic material. 1. Point mutations = base pair substitution (silent, missense, nonsense) 2. Frameshift mutations = Insertion or deletion of one or more nucleotide pairs
Silent
Fig 8.17
Mutations cont.
May be neutral (silent), beneficial, or harmful. Spontaneous mutation rate 10-6 1 mutation per million replicated genes Mutagens increase mutation rate 10 1000x
Chemical mutagens
Nucleoside (base) analogs have altered basepairing properties. They can be
randomly incorporated into growing cells (cancer drugs) only used by viral enzymes (e.g. AZT)
Fig 8.19a
Distortion due to intercalating agent will lead to one or more base-pairs inserted or deleted during replication.
Potent carcinogens!
Radiation as a Mutagen
1. Ionizing radiation (x-rays and -rays) lead to deletion mutations (ds breaks) 2. UV rays lead to thymine dimers (intrastrand bonding) Photolyases = light repair enzymes
(use energy from visible light to fix UV light damage)
Repair
Photolyases separate thymine dimers
Nucleotide excision repair
Fig 8.20
Test
Combine animal liver cell extracts with Salmonella auxotroph Expose mixture to test substance
Examine for signs of mutation in Salmonella, i.e. Look for cells (colonies) that have reverted from his to his+
Copyright 2006 Pearson Education, Inc., publishing as Benjamin Cummings
Fig. 8.22
Professor Richard A. Muller of UC Berkeley on the Ames Test and Natural Foods
Genetic Recombination
Vertical gene transfer: Occurs during reproduction between generations of cells. Horizontal gene transfer: The transfer of genes between cells of the same generation. Leads to genetic recombination.
Genetic Recombination
Exchange of genes between two DNA molecules Crossing over occurs when two chromosomes break and rejoin
Figure 8.23
1) Transformation
Naked DNA transfer
ANIMATION Transformation
Fig 8.25
2) Conjugation
Plasmid and chromosomal DNA transfer via direct cell to cell contact High efficiency F+ = donor cell. Contains F plasmid (factor) and produces conjugation (F) pilus (aka sex pilus) Recipient cell (F ) becomes F+ In some cells F factor integrates into chromosome Hfr cell R plasmids (R factors) are also transferred via conjugation
Copyright 2006 Pearson Education, Inc., publishing as Benjamin Cummings
Fig 8.26
Fig 8.27
ANIMATIONs
3) Transduction
DNA Transfer from donor to recipient cell with help of bacteriophage (= transducing phage) 2 types of phage-bacteria interaction: 1. Generalized transduction happens via lytic cycle caused by virulent phages
Fig 8.27
Transduction by a Bacteriophage