Module 6

Question 1

genetic engineering

Answer 1

manipulating the DNA or genetic material of an organism to include the DNA from a different organism

Question 2

plasmid DNA

Answer 2

small circular pieces of DNA that can replicate independently because they have an ori and can carry genes that encode for beneficial traits

easily transferrable

Question 3

transformation

Answer 3

process by which bacteria acquire altered genetic characteristics from a different source

Question 4

vector

Answer 4

serves as the vehicle for genetic material aka plasmid DNA

Question 5

reporter gene

Answer 5

makes it possible to visually distinguish bacteria that carry a plasmid with foreign DNA from those that don’t aka GFP

Question 6

competency

Answer 6

bacterial cell wall is made permeable to macromolecules such as DNA

can artificially be made competent by treating it with CaCl2 or abrupt transitioning between heat and cold environments

Question 7

pGLO plasmid

Answer 7

contains genes coding for amp resistance and for GFP production

Question 8

enzyme b-lactamase

Answer 8

secreted by transformed cells where it destroys ampicillin

Question 9

arabinose operon

Answer 9

set of genes that code for enzymes that break down arabinose

those genes were replaced by GFP gene

presence of sugar= genes will be transcribed and GFP will be produced

Question 10

ori

Answer 10

a sequence of DNA where replication is initiated

Question 11

selectable marker

Answer 11

a gene that presents resistance to ampicillin, resulting in the production of b-lactamase

Question 12

part 1

Answer 12

label one tube + and -

transfer 250 ul of CaCl2 into each tube and place both on ice

Question 13

pre-incubation

Answer 13

pick up a single colony of bacteria from starter plate and immerse it into transformation solution in both + and - tubes and place them back on ice

Question 14

incubation

Answer 14

get a loopful of pGLO DNA solution into + tube and mix it. none in - tube and then incubate tubes for 10 minutes on ice

Question 15

Plates

Answer 15

LB/amp: +pGLO
LB/amp/ara: +pGLO
LB/amp: -pGLO
LB plate: -pGLO

Question 16

Heat shock

Answer 16

put both tubes into water bath of 42˚C for 50 seconds then place them back in ice for 2 minutes

Question 17

recovery and growth

Answer 17

remove tubes from ice and add 250 ul of LB nutrient broth to both tubes and incubate for about 10 minutes at room temp

Question 18

Selection

Answer 18

pipet 100 ul of transformation onto each nutrient agar plates and use sterile loops and spread the suspension evenly
plates will be incubated for 24 hours

Question 19

Transformation efficiency

Answer 19

number of transformed colonies/amount (in ug) of DNA used x final transformation suspension (ml)/volume (ml) plated