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Announcements 1.Student-initiated genetics study group to meet Thursdays at 4pm in either Brooks 203 or 204 for problem solving. Review of Last Lecture 1. Finished linkage: mapping human chromosomes 2. Conjugation -F factor is a plasmid that allows for fertility and for a cell to act as a donor of genetic material F+ x F- F+ F+ F- F+ F+ Hfr is special F+ Hfr x FF- F+ Hfr F+ F- Outline of Lecture 15 I. Bacterial genetics, continued II. Extrachromosomal Inheritance Chloroplast DNA Mitochondrial DNA Maternal Effect Genes Circular Map of E. coli ~2000 genes Scaled in minutes, 100 minutes total Learning check A strain of Hfr cells that is sensitive to the antibiotic streptomycin (strs) has the genotype gal+ his+ bio+ pur+ gly+. These cells are mixed with an Fstrain that is resistant to streptomycin (strr) and that is gal- his- bio- purgly-. Cells are allowed to undergo conjugation. At regular intervals, a sample of cells was removed and conjugation was interrupted by placing the sample in the blender. The cells are plated on medium that contain streptomycin. Cells that grew were tested for the presence of genes transferred from the Hfr strain. Genes from the donor Hfr strain first appeared in the F- strain at the times listed. Give the order of the genes on the bacterial chromosome and indicate the minimum distances between them. gly+ 3 min. his+ 14 min. bio+ 35 min. gal+ 36 min. pur+ 38 min. Transformation: a different process of recombination, can be used to map genes * Relative frequency of genes being cotransformed indicates distance between the two genes Bacteriophages are viruses that use bacteria as hosts • Known as phages • Nobel prize awarded to Delbruck, Luria and Hershey in 1969 for their work on the genetic structure of phages T4 bacteriophage Virulent or temperate phage (e.g. phage) Lytic and Lysogenic Cycles Recombinants from transduction can be propagated Lederberg-Zinder experiment - Salmonella Could this recombination be due to conjugation or transformation? Transduction: virus-mediated bacterial DNA transfer Viruses can package bacterial DNA Transduction Recombination in Phage Large, dark recombinants Small, light • Strains with different plaque morphologies “crossed” by coinfection of bacteria: • h r + X h+ r – h mutant plaques are darker than h+ – r mutant plaques are larger than r+ • Results: parental (h r+ and h+ r) and recombinant (h+ r + and h r) plaques. Small, dark parental Large, light • # recombinants/total X 100% = recomb. frequency Learning check Given the following results of a cross between phage T2 strain A (h r+) and strain B (h+ r), determine the recombination frequency and distance between the h and r genes: Genotype Plaques h r+ h+ r h+ r+ h r 42 34 12 12 rII locus T4 Map From Recombination Analysis II. Extrachromosomal inheritance Some inheritance patterns are non-Mendelian and the phenotype is not transmitted by nuclear genes located on chromosomes of both parents II.A. Chloroplast DNA Chloroplasts contain their own genomes, are inherited maternally in the cytoplasm of the egg First example in 1908: Correns discovered branch leaf color in the four o’clock plant was affected by phenotype of ovule only, not pollen. Maternal Inheritance • Phenotype of maternal parent is solely responsible for phenotype of offspring • In Four o’clocks: – F white X M green > – F white X M variegated > – F green X M white > – F green X M variegated > – F variegated X M green > – F variegated X M white > white offspring white offspring green offspring green offspring variegated, green or white variegated, green or white Explanation of Results Variegated II.B. Mitochondrial Heredity • About 30,000 genes on 23 human chromosomes (3.3 billion base pairs/haploid cell) • Mitochondrial Genome contains 37 genes: – 13 code for some of the proteins involved in oxidative respiration – 22 tRNA genes – 2 rRNA genes – some proteins involved in mt protein synthesis – 16,569 base pairs, circular Human Mitochondrial Diseases • Are maternally inherited: only offspring of affected mothers are affected • Show deficiency in mitochondrial function • Are caused by a mutation in a mitochondrial gene • Example: myoclonic epilepsy and ragged red fiber disease (MERRF) – Deafness, dementia, seizures – Point Mutation in a mitochondrial tRNA Other Human Mitochondrial Diseases • Leber’s Hereditary Optic Neuropathy (LHON) – Sudden bilateral blindness – Point mutation in small subunit of NADH dehydrogenase • Kearns-Sayre Syndrome (KSS) – Symptoms in eyes, muscles, heart, brain – Deletion mutation in mtDNA Mitochondrial DNA and Diseases KSS LHON MERRF II.C. Maternal Effect: Snail Coiling • Pattern of offspring determined by genotype of mother, regardless of phenotype Although D is dominant to d, phenotypes of Dd and dd depend on mother’s genotype. Embryological Origins of Coiling Pattern: Orientation of Mitotic Spindles D product is deposited in cytoplasm during oogenesis. Maternal Effect Genes in Drosophila Development • Discovered by Edward Lewis, Christiane NussleinVolhard and Eric Weischaus (Nobel Prize in Physiology or Medicine, 1995) • Gene products deposited in egg by cells of mother’s ovary. • These genes set up the first embryonic axes: dorsalventral and anterior-posterior and control later zygotic genes.