eukaryotic gene regulation-6 Flashcards
name 3 different long range control elements
enhancers
LCRs
insulators
give the name of 3 different reporter genes
b-galactosidase
luciferase
gfp
what is the crucial control
the second plasmid should be co transfected
list some typical features of enhancer sites
DNase I hypersensitive
ATAC sensitive
H3K4me1
p300 binding
Med1 binding
binding of tissue specific transcription factors
what is the role of an insulator
block enhancer from contacting promoter
what reporter was used when characterising function of insulators
human y-globin promoter which leads to expression of neo
what was the enhancer that was used in this study
human b-globin LCR element-HS2
what is the second function of insulators
block the spread of chromatin
what reporter construct was used to find out this function
IL2R
what happened to the reporter gene when there were no insulators
it was repressed because of heterochromatin formation
how are transgenic mice generated
gene injected into male cell embryo
mother made pseudo pregnant
transgene randomly into genome
head to tail manner
what vectors are used to generate transgenic mice
plasmid
cosmid
BAC
YAC
name an advantage of gene expression via transgenic mice
in vivo transgene exposed to factors at correct development stage
analysis by reporter genes to see when promoters are activated
describe how the knock out mice method is done
isogenic transgene DNA put into ES cells
drug selection
surviving colonies screened for transgene
characterised targeted cells inject into mouse blastocysts
transferred to pseudo pregnant mouse
chimeric offspring identified
mated
to test transmission
what are the 2 targeting vector methods for knock out mice
gene targeting
random insertion
