BIOL SCI 215 Study Guide - Final Guide: Missense Mutation, Kras, Selectable Marker
28 Jun 2018
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6/1
Forward vs Reverse genetics
oForward genetics: Collect mutations that affect particular phenotypes and
testing
[Start with phenotype and look for the gene]
oReverse: Analyzing the phenotype following the disruption of the known gene
[Start with the gene and looking for the phenotype]
Gene phenotype interactions
Animal models, transgenesis
Neurogenetics
Role of genes in the development and function of the brain
Application of methods of molecular genetics to study the function of the brain
First study: Studying circadian rhythms in drosophila
Even if u put them in constant darkness, they still maintain a constant cycle of wake
and sleep
Forward genetic screen for circadian rhythm mutants uncovers period - gene which
times the biology/physiology of cells
Challenge: Complex genotype-to-phenotype interactions
Most life cycles are based on ubiquitination
One example is based on a real OCD Disease: thricotillomania
Look at SNP markings to find a region where the gene disease region could be
Sequence it to see if there is a mutation
Double selection
Use Knockout or CRISPR
How do you start an expression/ know which gene is expressed?
Study RNA
Northern Blot
Southern Blot - uses DNA [chopping by restriction enzyme]
Protein fusion: Go to the stop codon and before the stop codon, you put the GFP so that you
can fuse it to the protein
PCR: RT-PCR making RNA into cDNA which is a DNA copy and then u do a regular PCR
Base pairing to match but u do it in the animal
Midterm 3 Review
Restriction enzymes useful for making/cloning DNA
4 possibilities A,T,G or C, and there are 6 possible positions depending on number of
nucleotides, so possibilities to the power of positions
Human genome is 3x10^9
For insertions of plasmid products, you need the direction of insertion to be similar to
the ORF [ cos the mRNA coding information has to be the right way]
Building mouse knockout
Breeding to take heterozygous and make homozygous
If you are interested in dominant mutations, look at F1
find more resources at oneclass.com
find more resources at oneclass.com
Document Summary
Forward vs reverse genetics: forward genetics: collect mutations that affect particular phenotypes and testing. [start with phenotype and look for the gene: reverse: analyzing the phenotype following the disruption of the known gene. [start with the gene and looking for the phenotype] Role of genes in the development and function of the brain. Application of methods of molecular genetics to study the function of the brain. Even if u put them in constant darkness, they still maintain a constant cycle of wake and sleep. Forward genetic screen for circadian rhythm mutants uncovers period - gene which times the biology/physiology of cells. Most life cycles are based on ubiquitination. One example is based on a real ocd disease: thricotillomania. Look at snp markings to find a region where the gene disease region could be. Sequence it to see if there is a mutation. Southern blot - uses dna [chopping by restriction enzyme]
