insertion of a gene Flashcards
when we have staggered cuts what else do we have
we have sticky ends
the nucleotides at one end is what compared to the other
they are complimentry bases to each other
why
if the same restriction endonuclease is used to cut the fragments all will have the complementary bases`
why is it called sticky
as the single strand can join to another signle dna strand with complementry bases and form hydrogen bonds between adjacent bond pairs
what happens once the complementry bases come togetherq
tthe phosphate backbone must be restored using ligase
what is transcription
dna being copied into its complimentry MRNA strand
what needs to be added to our desired gene
a reigions which uses rna polymerase as well as a regiion which releases rna polymerase
what is the reigion that binds rna polymerase
it is called the protometer and also includes trnascripting factors
what happens does the rna polymerase do
it attatches to the nucleotide bases at the protometer and stimulate transcription
what does this rna polymerase allow
begin the process of transcription
what happens at the end
at the end terminator reigion rleases rna polymeras so the transcription stops and mrna strand is completed
once we have prepared our dna what nees to happen
it nees to be inserted into a vector like a plasmid
what is a plasmid
it is a circular length of dna that usually contains antibiotic resistance genes
what is done to the plasmid
the restriction endonuclease is used to cut a section of the dna to break the loop
what is done to the gene
using the same restriction enzymes section of dna required for deisred protien is cut
what happens to both ends
they both join with their sticky ends
how may the dna and the plasmid reformsation be ruined
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
why do plasmids make good vectors
as they are circular so are less likely to get brokem down
what else do they contain
marker genes in the form of antibiotic resistance which serve as recognition sites
why do we use reverse transcriptase
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
what does in vivo gene cloning mean
using a whole living organism and not just part of the bacteria
