Brainscape's Knowledge GenomeTM

Browse over 1 million classes created by top students, professors, publishers, and experts.


See full index

2014 Genetics Topic 3: RG > L5 > Flashcards

L5 Flashcards

You may prefer our related Brainscape-certified flashcards:
Biology 101 flashcards
1
Q

true or false, pUC vector allows propagation of molecule as ssDNA strand

A

true

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
2
Q

how to synthesize new genes in vitro

A
  • determine desired amino acid sequence
  • translate into desired DNA sequence
  • synthesize overlapping oligonucleotides
  • anneal together and ligate
  • clone assembled oligos into plasmid using sticky ends
  • sequence to confirm
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
3
Q

what methods are used to manipulate DNA sequences

A
  • cloning
  • PCR
  • sequencing
  • oligonucleotide sythesis
  • site directed mutagenesis
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
4
Q

how do you express high value proteins in fermentation

A
  • need high level promoter (to make lots of mRNA)
  • inducible expression- temperature shift, chemical induction (ethanol/methanol)
  • optimise translation- codon usage of host organism
  • optimise stability
  • optimise solubility
  • post translational modification
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
5
Q

How is DNA introduced in bacteria

A

electroporation or Ca2+ treatment

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
6
Q

How is DNA introduced in yeast

A

Li+ treatment or Electroporation

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
7
Q

How is DNA introduced in animal cells

A

Ca ppt of DNA
lipid encapsuation
microinjection

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
8
Q

How is DNA introduced in plant cells

A

PEG
electroporation
biolistics
A.tumefaciens

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
9
Q

true or false, if ori is present DNA replicated (all)

A

true

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
10
Q

if ori is absent and C/O is present what happens

A

DNA integrates by homologous recombination (10^-3 to 10^-4)

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
11
Q

what happens when ori is absent and there is a NHEJ

A

DNA integrates at random sites (10-3 to 10-4)

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
12
Q

true or false, results from with ori, C/O and NHEJ are stable transformants

A

true

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
13
Q

what does kanamycin do

A

inactivates aminoglycoside antibiotics (kanamycin, neomycin, gentamycin)

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
14
Q

true or false, kanamycin resistance is the most common selectable marker for eukaryote cells

A

true

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
15
Q

true or false, E.coli kanamycin resistance gene from Tn903: inactivates all of the mentioned antibiotics via phosphorylation

A

true

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
16
Q

where is genticin used

A

animal cells and yeast

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
17
Q

where is kanamycin used

A

plants

18
Q

what does genticin G418 inhibit

A

synthesis of cytoplasmic ribosomes

19
Q

what does kanamycin ihibit

A

chloroplast ribosomes

20
Q

true or false, eukaryote signals are used to express the kan gene

A

true

21
Q

in animals what is the promoter and the protein formed for kan gene expression

A

SV40 late promoter

SV40 poly A

22
Q

in plants what is the promoter and the protein formed for kan gene expression

A

CaMV 35S prom

CaMV poly A

23
Q

where are the signals derived from for kan gene expression

A

dsDNA of viruses, active in wide host range, direct high level of constitutive expression of transcript (can be turned on or off)

24
Q

what is a reporter gene

A

gene used to monitor level of expression from promoters

- no background activity in host cells

25
Q

what are common reporter genes

A

lac
gus
luc
gfp

26
Q

what is lac used for

A

bacteria, yeast, animals

27
Q

what is gus used for

A

plants

28
Q

what is luc used for

A

yeast, animals, plants

29
Q

what is gfp used for

A

yeast, animals , plants

30
Q

true or false, lac and gus are standard reporter genes

A

true

31
Q

true or false, luc is the most sensitive reporter gene

A

true

32
Q

true or false, gfp is a non destructive reporter gene

A

true

33
Q

true or false, yeast is the best model eukaryote for gene transfer

A

true

34
Q

why is yeast the best model for eukaryote gene transfer

A

has set of vectors for many purposes

  • integration
  • high copy plasmid
  • low copy plasmid
35
Q

what are the commercial applications of yeast

A

hepatitis B vaccine

fermentation

36
Q

What are the two kinds of integration events

A

1.) random:
occur anywhere on any chromosome ( by DNA repair systems called NHEJ: non homologous end joining, no DNA homology needed)
- allows gene addition but not gene replacement
- only dominant genes produces a phenotype
- variable expression
2.) targeted:
occur at sites of sequence identity (by homologous recombination system –> control of integration site)
- allows gene replacement as well as addition
- both dominant and recessive genes produce phenotypes, including null alleles)

37
Q

true or false, targeted integration is the most powerful system

A

true

38
Q

true or false, targeted integration is predominant in yeast

A

true

39
Q

how do you do target integration

A

via homologous recombination to replace ORF in yeast

40
Q

how do species without targeting integration reduce gene expression

A

by introducing dsRNA , inductions of a sequence specific nuclease that specifically degrades that target RNA, results in dominant negative transgene

2014 Genetics Topic 3: RG (10 decks)

Brainscape helps you reach your goals faster, through stronger study habits.
© 2024 Bold Learning Solutions. Terms and Conditions