definitions Flashcards
restriction enzymes function
is a protein isolated from bacteria that splices DNA sequences at sequence-specific sites, creating DNA fragments with a known sequence at the end
where do restriction enzymes come from
naturally occurring in bacterium/being able to restrict viruses
what is a palindrome
same forwards and backwards on both strands of DNA
why is a palindrome important to restriction enzymes
allows enzyme to know where to cut, can recognize sequence on both strands, and insures that both strands are cut
what are sticky ends
when another type of restriction enzyme cuts a DNA sequence to create two uneven ends that connect back to one another easily (puts fragments together easy)
what are blunt ends
when a certain restriction enzyme separates a DNA sequence to create two ends of DNA which are equal on both sides (keeps fragments together)
DNA ligase function
attaches and rejoins DNA bases with complementary bases
recombinant DNA (DNA has been recombined)
fabricated by combining different parts of an organism into one (DNA that has been cut and pasted)
gel electrophoresis
separates large DNA fragments from small DNA fragments
why do DNA fragments travel through the gel
DNA is negative, when the machine is turned on the positive magnets pull the fragments through the gel
speed of small fragments
travel through gel faster because there is less of it
speed of large fragments
travel through gel slower because there is more of it
why do we have size standards
to compare the lengths of the fragments (measured in base pairs)
transforming bacteria
putting plasmid into bacteria
ways of transforming bacteria
shock, poison, cook
steps of cloning a gene
- cut plasmid and gene of interest
- mix plasmid and gene of interest
- dna ligase seals them
- transform (shock, poison, cook)
- it replicates during cell division
polymerase chain reaction
A PCR is a method that allows DNA copies to be made!!
It uses a thermal cycler (PCR Machine)
why do we use PCR
allows us to create/duplicate DNA strands relatively easily compared to other methods.
steps of PCR
- heat: denature-creates two strands
- build dna: annealing- primers add or “attach” a short piece of single stranded complementary DNA to both ends of the original DNA strand.
- taq polymerase adds free floating nucleotides to each strand,
function of primers
they prepare or “prime” the site where taq polymerase can start working. This also ensures that either sides of the target region are copied on the Template DNA.
function of taq polymerase
adds free floating nucleotides to each strand
function of dna sequencing
tells us the sequence of the strands
genetic engineering
cutting and pasting dna
how can genetic engineering make GMO’s
-viral vectors
-gene gun
viral vector
transfers gene to modify cell tissue virus delivers DNA straight to chromosomes
gene gun
delivering genetic material into plants
how can plasmids and viruses be used as vectors
plasmids: can be placed in prokaryotes
virus: can deliver recombinant dna in eukaryotes
application for GMO
-bacteria that can make human insulin and covid vaccine (bacteria makes covid protein)
-eukaryotic gmo: enviro pig that poops clean
-recombinant plasmid into vector: dna can get into a host cell (cell makes holes bc of stress)
recombinant virus into vector: delivers dna
gene therapy
treats/cures genetic disease
reproductive cloning
creation of a genetically identical organism
therapeutic cloning
source of embryonic stem cells
somatic(body) cell nuclear transfer
nucleus from somatic cell is transferred to enucleated oocyte
(copy gene of interest) gene of interest
you can make copies/get the protein
(copy gene of interest) origin of replication
the plasmid is going to get copied
(copy gene of interest) antibiotic resistance gene
bacteria who have the plasmid will live
(make the protein) promoter sequence
transcription starts
(make the sequence) terminator sequence
transcription stops
multiple cloning sequence
allows DNA sequence to be inserted without messing up the plasmid
T7 pro
regulates gene expression of recombinant proteins
