NEURO410 Lecture Notes - Lecture 6: Northern Blot, Knockin, Homologous Recombination

• School: University of Alberta
• Department: Neuroscience and Mental Health Institute
• Course: NEURO410
• Professor: Dr.Kar

Document Summary

Lecture 6: trans-genics and knock-outs: use in aging and ndd research (jan 21/16) Classical genetics and basic dna engineering: dealing with genes one at a time. Genetics refresher: the crucial concept of gene copy-number. Gene copy number = relevant to way we make transgenic mice/knock in/out mice. Ba(cid:272)teria are (cid:862)haploid(cid:863) and generally have one copy of each gene. Hu(cid:373)a(cid:374)s are (cid:862)diploid(cid:863) and generally have two copies of each gene (some exceptions, some genes are repeated ie ribosomal proteins) Dominant mendelian only one copy in an (cid:862)allele(cid:863) genome is altered. Recessive mendelian (cid:271)oth (cid:272)opies i(cid:374) a (cid:862)diploid ge(cid:374)e(cid:863) genome needs to be altered. Do(cid:373)i(cid:374)a(cid:374)ts are usuall(cid:455) a (cid:862)gain of pathologi(cid:272) fu(cid:374)(cid:272)tio(cid:374)(cid:863) Ingredients for pronuclear microinjection: injected via a needle into a mouse. Correctly assembled dna for the transgene, a (cid:862)construct(cid:863): small cassette of dna that does what we want ie make a protein. Transgene specific probes for proving correct mrna expression (northern blot, rna protection assay, rt-pcr)