insertion of a gene

Question 1

when we have staggered cuts what else do we have

Answer 1

we have sticky ends

Question 2

the nucleotides at one end is what compared to the other

Answer 2

they are complimentry bases to each other

Question 3

why

Answer 3

if the same restriction endonuclease is used to cut the fragments all will have the complementary bases`

Question 4

why is it called sticky

Answer 4

as the single strand can join to another signle dna strand with complementry bases and form hydrogen bonds between adjacent bond pairs

Question 5

what happens once the complementry bases come togetherq

Answer 5

tthe phosphate backbone must be restored using ligase

Question 6

what is transcription

Answer 6

dna being copied into its complimentry MRNA strand

Question 7

what needs to be added to our desired gene

Answer 7

a reigions which uses rna polymerase as well as a regiion which releases rna polymerase

Question 8

what is the reigion that binds rna polymerase

Answer 8

it is called the protometer and also includes trnascripting factors

Question 9

what happens does the rna polymerase do

Answer 9

it attatches to the nucleotide bases at the protometer and stimulate transcription

Question 10

what does this rna polymerase allow

Answer 10

begin the process of transcription

Question 11

what happens at the end

Answer 11

at the end terminator reigion rleases rna polymeras so the transcription stops and mrna strand is completed

Question 12

once we have prepared our dna what nees to happen

Answer 12

it nees to be inserted into a vector like a plasmid

Question 13

what is a plasmid

Answer 13

it is a circular length of dna that usually contains antibiotic resistance genes

Question 14

what is done to the plasmid

Answer 14

the restriction endonuclease is used to cut a section of the dna to break the loop

Question 15

what is done to the gene

Answer 15

using the same restriction enzymes section of dna required for deisred protien is cut

Question 16

what happens to both ends

Answer 16

they both join with their sticky ends

Question 17

how may the dna and the plasmid reformsation be ruined

Answer 17

if the plasmids own sticky ends cause the plasmid to close up before the incorporation the dna segment
or the dna segment join together to form their own plasmid

Question 18

why do plasmids make good vectors

Answer 18

as they are circular so are less likely to get brokem down

Question 19

what else do they contain

Answer 19

marker genes in the form of antibiotic resistance which serve as recognition sites

Question 20

why do we use reverse transcriptase

Answer 20

as the resulting dna does not have introns, we don’t have to locate the genes and it forms a stable copy of the gene

Question 21

what does in vivo gene cloning mean

Answer 21

using a whole living organism and not just part of the bacteria