MCB 160L Overview•Forward genetics: Define a phenotype and screen or select for mutations that generate that phenotype, starting with a random mutant collection (Arabidopsis-gametogenesis, embryonic phenotypes; Drosophila-specific gene expression patterns)….mutagen = mobile DNA elements. Goal-Move from knowledge of the phenotype to identification of the gene. •Reverse genetics: Start with a specific gene, identified at the molecular level. Goal-Create or identify mutations in that specific gene in order to study the phenotype produced when gene function is reduced or removed. (C. elegans–RNAi to produce a phenocopy of mutation; S. cerevisia(yeast)–targeted knockout by homologous recombination) •Mapping the location of genes:Analysis of segregation and linkage to determine chromosomal location; 3-pt cross; molecular and bioinformatic tools•Molecular genetics tools:DNA isolation, restriction mapping and agarose gel electrophoresis, plasmids and bacterial transformation, subcloning and genetic engineering, PCR (as a tool for construction and analysis of DNA sequences, as a method to analyze genotypes obtained from small tissue samples, as a genomics tool)
has intentionally blurred sections.
Sign up to view the full version.