Yeast biotechnology 2

Question 1

what allowed the screening of role of each gene in a yeast phenotype and what was found

Answer 1

systematically knocking out all of the genes in yeast

only 15% of the genome is essential

Question 2

when knocking out the genes, what was done and why

Answer 2

a unique molecular barcode was incorporated - each deletion was tagged with one
so that you could tell which mutants were doing well under different conditions - can screen for antibiotic resistant genes

Question 3

what is haploinsufficiency

Answer 3

half the amount of gene product

Question 4

what type of yeast does mating only occur in

Answer 4

hapoloids, which can be either a or alpha, as they display the same sexual differentiation

Question 5

what are snowflake yeast

Answer 5

advantage to any yeast that makes it replicate in groups - rotifers don’t like them
- leads to the evolution of multicellularity

Question 6

what is an interactome

Answer 6

protein-protein interactions - complexes

Question 7

what is synthetic lethality

Answer 7

2 mutations together makes a much stronger phenotype so the yeast dies
these proteins may work in the same complex/pathway and have similar functions

Question 8

what is URA3 and what does it mean for the cell

Answer 8

selectable genetic marker in yeast, they can’t make Uracil so die unless they have uracil in the medium or a URA3 plasmid

Question 9

what happens if you add 5-FOA into yeast cells expressing URA3

Answer 9

they die

Question 10

what is special about using URA3, and how is this possible

Answer 10

you can select for and against URA3
as yeast cells without it die
but yeast cells with it can be killed when 5-FOA is added

Question 11

what is vip1delta

Answer 11

very important protein when looking for synthetic lethal phenotype

Question 12

what TAP-tagging and the main benefit if this method

Answer 12

Tandem Affinity Purification - allows protein purification under mild conditions = no harsh chemically needed - protein remains intact not broken up like in previous methods

Question 13

what is the core proteome

Answer 13

basic essential gene functions for survival - covering basic eukaryotic cell functions for life

Question 14

what is synthetic yeast

Answer 14

all DNA is synthetic - made on computers

want to create a unique yeast but optimising codon usage, removing waste eg repeats