PLBI Lab Quiz 3 - Lab 4 and 5 Flashcards
(37 cards)
genus and species name for arabidopsis
arabidopsis thaliana
which family is arabidopsis from
brassicacae
arabidopsis is related to which food groups
canola
why is it ideal for plant biology research
small, fast growth, self pollinating, small genome
what are transgenic arabidopsis lines
lines that have been genetically engineered to express a new trait not naturally found in them
in arabidopsis, what is the foreign gene that modifies arabidopsis, and how is it integrated
the fluorescent protein marker, its it’s integrated into the genome so every cell in the transformed arabidopsis will express the fluorescent protein marker
how is cell plasticity useful to us
allows regeneration of a whole plant from a single cell, so the single tranformed cell will be able to generate a whole plant that expresses the protein
out of all these organelles, which ones express GFP vs RFP, and why are they labelled
mito, golgi, er, tubules, peroxisome - all GFP, so we can visualize their movemnts and morphology easily
What do we use epifluorescence techniques to do in the lab (general)
to visualize fluorescent labelled structures within cells
in this lab (4), what are we using epifluorescnece microscopy to visualize
cytoskeleton, organelles in root epidermal cell of arabidopsis
name 2 challenges of autofluroescence
Background fluorescence in tissues (like leaf mesophyll cells) interferes with imaging due to chlorophyll and cell wall fluorescence.
Root epidermal cells are less autofluorescent, they’re more suitable for detailed imaging with less background interference.
arabidopsis lines are genetically engineered to express flurorescence under which protein
GFP
For the targeting mechanisms, organelles are labelled via [blank] or [blank]
targeting signals or binding proteins
example of targeting signal and organelle
SKL peroxisome targeting sequence (for peroxisomes)
example of binding protein and organelle
MAP 4 microtubule binding protein for microtubules
high or low power brightfield is used to visualize cytoplasmic outer layer
low
Why do more cells fluoresce in the transgenic Arabidopsis lines compared to bombarded onion cells?
foreign gene encoding GFP is stable in arabidopsis, but in onions, theres only a small localized region to express the gene so less fluroescence
w
why is there more autoflurescnece in leaf tissue than root tissue
Leaf tissue contains chlorophyll, which has strong natural fluorescence under epifluorescence microscopy, leading to significant background signal.
Root cells lack chlorophyll, resulting in minimal autofluorescence and clearer imaging of labeled structures.
Are the organelles in Arabidopsis root cells similar in morphology to those in onion leaf cells?
yes, similar morphology and comparable functions. but organization and distribution differ
What does immunofluoresence microscopy do
visualize specific proteins, target them with antibodies conjugated to fluorescent proteins
difference between epiflurosences and immuno in terms of localization
fluorescent = mislocalization, immunofluorescence = accurate localization proteins.
lab 5 uses [blank] (direct or indirect) immunofluorescence, which involves [blank] antibodies to amplify the fluorescent signal for clearer imaging.
indirect, primary and secondary
Primary Antibody
recognizes protein of interest, monospecific to reduce BG noise
Secondary Antibody
conjugated to flurosecnet molecule, bind to primary antibody for enahnced visibility
