application of genetic engineering methods Flashcards
give 2 froms of target identification
microarrays and yeast-2-hybrid screen
what does microarrays compare?
transcribed genes in 2 different tissue, or 2 different conditions of same tissue
how are microarrays formed?
in a cell free system using isolated mRNA
what does probe cell contain?
many copies of 25 base sequence that matches a gene
what is mrna tranformed into in microarray?
ssDNA
how do you find out what regulates a certain protein?
you look for proteins that physically interact with it
what are 2 proteins bound to in yeast - 2 -hybrid screen?
A is bound to binding domain
B is bound to activation domain
how do you find function of certain proteins?
observe effects of its absence
give 2 ways of a gene knockout
siRNA and using transgenic mice
what pathway does sirna use?
RNA interference pathway
give 3 steps of siRNA mediated gene knockdown
foreign double stranded RNA introduced into cells
dicer cleaves RNA
RISC degrades target mRNA
what does risc stand for?
RNA-induced silencing complex
give 2 methods of producing transgenic mice
random integration + homologous recombination
why do chimeric mice produce 2 types of gamete?
1 from host blastcyst, one from injected ES cells
