Lecture 7 Flashcards
(10 cards)
What is gene cloning?
A form of recombinant DNA technology which isolates sections of DNA and creating clones of them
How is DNA copied? (short answer)
Cut DNA using restriction enzymes from bacteria into plasmids and insert into the bacteria
The bacteria will copy itself and the DNA
What is a cloning vector?
A DNA molecule that can be introduced into a host organism and can self-replicate
What are restriction enzymes?
They cut the DNA into fragments by recognising specific nucleotide sequences in DNA and cut both strands of the sugar-phosphate backbone
What are the types of bacteria restriction enzymes?
Hind III
BamHI
Alu I
Sau3 AI
How can we find out how close sites/genes are in relation to each other?
We can use pairs of specific enzymes to cut at specific regions and then find the size of the DNA fragment by using gel electrophoresis
How can we use eukaryotic protein in therapies?
We can’t use them on their own as they are too big and so we isolate the mRNA by removing introns and convert this into DNA (since we cannot put mRNA into a plasmid) to create cDNA
What are expression plasmids?
Vectors that allow us to express clone genes of proteins
What are eukaryotic expression plasmids?
They introduce cloned DNA to bacteria to allow expression of the clones gene
What are the steps of the Southern blot?
DNA fragments are separated by gel electrophoresis and denatured by soaking the gel in alkali
DNA fragments are transferred onto a DNA binding membrane
DNA probe is place in hybridisation solution with membrane
DNA hybridises to the complementary DNA fragment on the membrane
Position of complementary DNA fragment can be identified as it is labelled by the probe
DNA gel put into nuclei and paper towels placed on top
DNA dragged out of the gel and onto the membrane
Radioactive/Fluorescent probe is added and will only pair to one specific restriction fragment on DNA
Probe is washed off and DNA is placed under X-ray, now we can view the size of the DNA and clone it
