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Today’s Agenda: 1. A Microarray Primer 2. Guest Speaker: Dr. Michael Schlador 3. Follow-up: the Use of Microarray Analysis in Chemotherapeutics 4. Preview of Chapter 15 & 16 Chapter 15: Mutations • Mutations generate genetic variants • These variants are then subject to recombination Consequences of mutations What would you predict if the mutations was….. • Within a coding region • Within a noncoding region What are the effects of point mutations on gene products? Are mutations random or induced? • Luria and Delbruck used the “fluctuation test” • They were interested in determining which hypothesis was correct Replica plating What can cause spontaneous mutations? • Errors in DNA replication – Transitions: base pairs mismatch – Transversions: base pair mismatch with the normal keto base forms What can cause spontaneous mutations? • Errors in DNA replication – Frameshift mutations: Also known as indel mutations Trinucleotide repeats may lead to disease…why? How can we induce mutations? • Use base analogs – Cause incorrect base pairing How can we induce mutations? • Use intercalating agents which mimic base pairs and slip in between the base pairs…promote indel mutations How can we induce mutations? • Promote base damage – UV light – Ionizing radiation • Promotes large strand breaks – Aflatoxin B1 • Binds to guanine and generates an apurinic site Ames Test is to determine mutagenicity Biological Repair Mechanisms • Direct reversal of damaged DNA • Base-excision repair • Nucleotide-excision repair • Mismatch repair Direct reversal of damaged DNA Base-excision repair • Carried out by DNA glycolylases, generate apurininc or apyrimidinic sites • AP endonuclease nicks strand • Deoxyribophosphodiesterase removes more DNA • DNA polymerase fills in the gap with new DNA Nucleotide-excision repair • Used to repair base damage or transcription blocks • Autosomal recessive diseases, Xeroderma pigmentosum and Cockayne Syndrome result from defects in this system Mismatch repair • Recognize mismatch base pairs • Determine which base is the incorrect one • Excise the incorrect base and repair DNA Error prone repair • Known as translesion synthesis, requires a bypass polymerase Repair of double-strand breaks • What conditions can cause double strand breaks? • There are two possible situations: – Nonhomologous end joining – Homologous recombination Nonhomologous end joining Homologous recombination • Damage corrected by synthesis-dependent strand annealing (SDSA) • Uses sister chromatids as templates Why can mutations lead to cancer? • Two types of mutations associated with cancer – Oncogenes are activated – Tumor suppressor genes are inactivated Oncogenes • Genes that control the cell cycle or inhibit apoptosis are considered protooncogenes Tumor Suppressor genes • Often these genes participate in regulation of cell cycle, activate cell apoptosis, or repair of damaged DNA • Mutations in p53 result in 50% of all tumors Chapter 16: Changes in chromosomes Chromosome numbers • Changes in chromosome sets are known as aberrant euploidy • Changes in parts of chromosome sets is known as aneuploidy Polyploids • More common in plants • Correlation between the number of chromosome sets and size of organism • Autopolyploids: multiple chromosomes from one species • Allopolyploids: sets of chromosomes from two or more different species Agricultural applications • Monoploid plants provide a way to select for desired traits Agricultural applications • Bananas are sterile triploids • Autotetroploid grapes are larger than diploid grapes Aneuploidy • Mostly due to nondisjunction during meiosis or mitosis Abnormal numbers of sex chromosome • Turner Syndrome – Result of only one sex chromosome (XO) Abnormal numbers of sex chromosome • Klinefleter Syndrome – The result of an extra X chromosome (XXY) Abnormal number of autosomes • Down syndrome – Results from an extra copy of chromosome 21 Can Down syndrome be inherited? Cancer from translocation Readings for Chapter 15/16 • Chapter 15: all sections except 15.5 • Chapter 16: section 16.1, 16.2 Robertsonian translocation (584-585) and rearrangements and cancer (587)