Manipulating genomes Flashcards
(33 cards)
What is the definition for DNA sequencing?
Finding the nucleotide sequence for gene or the whole genome
What are the 3 steps of Sanger sequencing?
1)Create copies of DNA fragments
2)Create complementary DNA fragments
3)Analyse complementary DNA fragments (by length)
For Sanger sequencing How do you create copies of DNA fragments? (4)
Extract DNA sample
Heat to form 2 strands
Cut strands into fragments
Copy strands
For Sanger sequencing How do you create complementary DNA fragments ?
+ fragments into mixture of DNA nucleotide DNA polymerase, DNA primers and terminating DNA nucleotide (stops+more nucelotides)
DNA polymerase joiners primers and terminating nucleotides
What are the 3 benefits of using High-through put sequencing?
its automated
Rapid
cheaper
What are the 3 main benefits of DNA sequencing?
1)Genome wide comparisons w individuals and species reveal relatedness
2)Prediction of amino acids sequences reveals tertiary structure of polypeptides
3)Synthetic bio - developing drugs
What is Gel electrophoresis used for?
To separate:
DNA- by length/mass
RNA - by length/mass
Proteins - by mass/Charge
How would you set up a Gel electrophoresis? (3)
1)Cut wells into a piece of Agar gel and submerse it in buffer solution
2)Load subjects into wells and place an electrode at each end
3)Apply current and watch+time how molecules move from negative to positive electrode
In Gel electrophoresis, what does it mean if a molecule moves further down/faster than others?
The molecules is lighter/shorter/more negatively charged than the others
Suggest what one could do to help differentiate between wells?
+ Fluorescent dye which glows under UV light
What is the definition for genetic engineering
Isolating a gene from one organism and placing it in another
What is the definition for gene therapy?
When a patients DNA is altered to treat or cure a disease
What is a somatic cell?
A body cell expect for germ cells (sperm and egg)
What is Somatic gene therapy?
This is when a new gene is added to a somatic body cell to directly target tissue that needs treatment
What are the flaws with somatic gene therapy?
The effects it provides are short lived as eventually the somatic cell will die
What is Germ-line gene therapy?
A Gene is implanted into a sex cell so the gene wil be inherited in birth
-so long term effects
What are 3 uses for genetic engineering?
1)Modifying plants to become insect resistant
2)Modifying pathogens to find new medical treatments
3)Gaining new proteins from animals and medicine testing on animals
What are some ethical issues raised by genetic engineering?
“should we modify animals”
“should we put human genes in animals at all”
Patents being money hungry type shit
How is restriction endonuclease used to produce DNA fragments?
They break the phosphodiester binds between adjacent nucleotides at the recognition sequence
-They either cut straight or leave overhang which creates a DNA fragment
How is reverse transcriptase used to produce DNA fragments?
1)Reverse transcriptase +s DNA nucleotides to mRNA
2)This forms a new strand of cDNA
3)An enzyme destroys mRNA
4)DNA polymerase +s nucleotides to the cDNA and now a DNA fragment is formed
Why do scientists use vectors when implanting a gene?
So the gene is not seen as foreign and thus rejected
It will then carry the DNA fragment into the host cell
How do you insert a DNA fragment into a plasmid to form a recombinant plasmid? (3)
1)Use restriction endonuclease to from hang over DNA fragment
2)Use the hang over fragment to cut the plasmid so they are complimentary
3)H-bonds a DNA ligase bind the nucleotide together and form phosphodiester bonds = recombinant DNA
Why cant plasmids regularly join to a host cell? (3)
Cuz they are polar, negatively charge and hydrophilic
What is the definition for transformation?
The process of adding a plasmid to a host cell
