L3

Question 1

primers

Answer 1

all same melting temp, not self complementary can cause weird side products

Question 2

plasmids

Answer 2

Autonomously replicating extrachromosomal DNA element, dsDNA circle, Plasmids can harbor genes picked up via DNA recombination or
transposition; they can be transmitted vertically to daughter cells upon division, or
horizontally to other bacteria (complex)

Question 3

theta replicating placmid

Answer 3

● Synthesize RNA primer, primer elongation with dNTPs
● At each replication fork, one strand is continuously synthesized and
the other discontinuously via Okazaki shite
● Uni or bidirectional

Question 4

strand displacement placmid

Answer 4

● Each strand has its own origin
● Helicase melts A-T rich section
● Primase makes two RNA primers
● DNA poly 3 elongates dNTPs
● There is NO LAGGING STRAND

Question 5

rolling circle placmid

Answer 5

● Relaxase breaks the strand, the resulting 3’ OH serves as primer
● The strand kind of unrolls
● The complementary strand is synthesized from an origin
● These are replication intermediates of bacteriophages and archaeal
plasmids

Question 6

important plasmids

Answer 6

● pAW63 (bacillus cereus): studied for war, it’s a bio weapon
● pBluecript plasmid: For DNA manipulation detection

Question 7

type iv secretion system

Answer 7

contain a channel through which proteins or protein–DNA complexes can be translocated
● Only DNA that can be recognized by the type IV protein is transferred
● If the plasmid is part of the chromosome, the entire chromosome can be
transferred into a new host.

Question 8

cloning recombinant dna

Answer 8

● Cut the desired DNA with an endonuclease
● Cut the plasmid vector with endonuclease
● Check whether the DNA was cut at the right sites
● Paste the two DNA using ligase (this is the recombinant DNA)
● Transform the DNA into E. coli
● Select E. coli bearing the DNA

Question 9

restriction enzymes

Answer 9

● Very binding/cutting site specific
● Cut in palindromes
● They restrict the activity of phages
● They avoid cutting their own DNA by adding methyl groups, to which the
enzymes cannot attach to

Question 10

plasmid cloning limitations

Answer 10

dna can break by shaking

Question 11

lambda fages

Answer 11

insert genome into e coli, 8-23 kb dna fragments

Question 12

BAC

Answer 12

inserts between 50-350 kb, derived form F plasmid they code for proteins for partitioning of chromosomes in daughter cells, it replicates up to 350 k bp, ligations made in gel to avoid framentation, transformation with electroporator (works only non k12 e coli stians)