lab 3 Flashcards
why do we use mutation of glp 1gene
it sterilizes the worms so they don’t lay eggs. this is important because if they do lay eggs, the eggs will grow and then we won’t know if a worm is one that we are trying to study or if it’s one of the new worms.
how does glp 1 mutant work at high and low temp
low temp: a missense mutation/change in amino acid, and this doesn’t affect protein function
when worms are at high temp, mutation leads to a missense mutation that leads to protein misfolding and loss of function
what are we doing for exp 1 this week
picking and imaging the worms that we placed on L440 (no knockdwon) and L440YFP (YFP knockdown) last week. We will image the worms (YFP should be seen on L440 and less on L440YFP) and measuring worm speed: should worm speed differ between the plates?
genetic screen
population of cells or roganisms w a variety of random mutations, overexpression or repression of genes, etc
why do we use genetic screens
the organisms have a variety of gene mutations/differ from each other. the population of organisms is used to identify the genetic deviation that resutlts in a desired pheontype.this allows us to find new genes involved in a particular cell function
why is genetic screening an unbiased method
bc we don’t know what genes are involved in a particular function- cell tells us
if a worm has a gene knocked down and that worm was able to survive heat shock, what does that say about the gene knockdown?
gene knockdown enhanced proteostasis network. that’s why worm could survive heat shock.
why are we knocking down GOI?
To see if the knockdown enhances PN.
why do worms look green even though we’re using yfp
computer thinks we’re using gfp bc the excitation and emission wavelengths of gfp and yfp are v similar
diseased worms are
glp1-q35-yfp worms
mutation of glp 1 gene is always present in genome: T OR F
T
what is the glp 1 gene
gene whose protein product is essential for growth of egg and sperm
what causes unfolded proteins
mutation (like glp 1 mutant), stress, heat, etc
PN solutions
turn down translation, turn up chaperone expression/protein degradation, autophagy, ER/mito unfolded protein response (can turn on chaperones and other mechanisms)
what can lead to diseased state/lower PN
aging, protein damage, energetic deficits, biosynthetic errors, and mutations
