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Lecture 11

Lecture 11 - Generating Mutants

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Biology 2581B
Jim Karagiannis

LECTURE 10: GENERATING MUTANTS Generating Mutants • Early on, we relied on mutants which either occurred spontaneously or were induced – random • Nowadays, we like to generate mutants with a specific mutation in a specific place, if possible “Early” Mutants • Often morphological mutants • Often spontaneous • Arabidopsis thaliana o 1873 – first documented Arabidopsis mutant byAlexander Braun o 1990 – this gene was actually cloned and characterized • Drosophila melanogaster o 1910 – discovered by Thomas Hunt Morgan and Lilian Vaughan Morgan o First sex-linked mutation ever discovered in the fruit fly • Often morphological mutants are typically spontaneous in origin • Mus musculus o Brachyury mutation: Greek brakhus – short oura – tail o 1927 – described by Nadine Dobrovolskaïa-Zavadskaïa o Heterozygous: affects tail length and sacral vertebrae o Homozygous: lethal at around embryonic day 10 due to defects in mesoderm formation, notochord differentiation and the absence of structures posterior to the forelimb o Nude mice (1962 – discovered by N.R. Grist in Glasgow) o Lack a thymus and cannot generate mature T lymphocytes. Therefore, they are unable to mount most types of immune responses o The genetic basis: disruption of the FOXN1 gene • C. elegans o 1974 – first suggestion to use this as a model organism • Problem: spontaneous – too few – too slow and wee need mutants other than morphological in biochemical pathways, with subtle changes o Induced mutations = increased frequency and larger variety Inducing Mutations • Popular mutagen • EMS ethyl methanesulfonate alkylating agent EMS • Popular mutagen • Induces point mutations – base pair substitutions • GC toAT transitions • Ease of use: soluble, can be taken up by cells • You start to screen individuals for morphological phenotypes, biochemical mutants, etc. • You can use assays to screen individuals for developmental mutants, for mutants with different behavior patterns, etc. • You build a collection of mutants, and if you identify mutants with the same phenotype, do they carry a mutation in one gene or several genes? • Problems o Random mutations o No selection o Often more than one mutation per mutant o Single base pair change somewhere in the
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