Recombinant DNA (Transgenics) Flashcards
Describe techniques and applications used in recombinant DNA technology for example: - The development of transgenic organisms in agriculture.
What is recombinant DNA?
DNA that is formed artificially as it contains genes from two or more different sources
What is recombinanat DNA technology?
(1) methods to join together DNA from two different species (2) in order to produce new genetic combinations and (3) allow the organism that the gene has been transferred to to express the given genes to make protein
What is the scientific process involved in recombining DNA?
isolating DNA fragments from one genome using restriction enzymes, which can cut
DNA at a precise sequence of bases.
What do restriction enzymes do in the recombinant DNA process?
isolating DNA fragments and cutting them at a precise sequence of bases
What does DNA ligase do in the recombining DNA process?
the rejoin the cut DNA pieces
What is ligation?
reassembling DNA after it has been cut by restriction enzymes
What is step (1) of the recombinant DNA technology process?
The required gene is isolated from a
cell.
What is step (2) of the recombinant DNA technology process?
a piece of circular DNA (called a plasmid) is removed from bacteria
What is step (3) of the recombinant DNA technology process?
two pieces of DNA (one from the top strand and one from the bottom stand) are cut from the plasmid using the restriction enzyme
What is step (4) of the recombinant DNA technology process?
both the cut parts of the plasmid and the isolated gene have sections of single-strand DNA with exposed nucleotide bases (called sticky ends) that match witht he other strand
What is step (5) of the recombinant DNA technology process?
the bacterial plasmid is cut at two points using the same restriction enzyme
What is step (6) of the recombinant DNA technology process?
the sticky ends of the isolated gene and the plasmid come together, and join up via base pairing. The whole process of this step is called annealing
What is step (7) of the recombinant DNA technology process?
the enzyme DNA ligase joins the DNA fragments to form a circular plasmid/linear molecule
What is step (8) of the recombinant DNA technology process?
The plasmid is inserted back into a
bacterial cell, where multiple copies
of the gene can be produced.
What can be done with the multiple copies of the gene after they are produced?
the gene can be
inserted into an egg cell of another
species and, after fertilisation, becomes
part of the newly formed organism’s
DNA
What is step (6) of the recombinant DNA technology process?
As the sticky ends of the human gene and the plasmid
What is a plasmid
a piece of circular DNA, made up of two DNA strands
Where can you find a plasmid?
in a bacteria
How many methods are there of inserting a desired gene into the genome of a species?
4
What are the names of th four methods of inserting a desired gene into the genome of a species?
M.I.B E.T
1. Micro injection
2. biolistics
3. electroporation
4. transduction
What method of inserting a desired gene into an organism’s DNA involves injecting the DNA directly into the nucleas of a single cell?
micro-injection
What is one way microinjection is actually used?
to introduce DNA into egg cells when creating transgenic species
What method of inserting a desired gene into an organism’s DNA involves mechanically delivering DNA on microscopic particles into target tissues and cells by “firing”/”shooting” the DNA from a gene “gun”?
biolistics
How can biolistics be used in real life?
tiny gold particles can be used to coat the DNA, which is then
fired at the target cells under high pressure or voltage
What method of inserting a desired gene into an organism’s DNA involves increasing the membrane permeability by applying an electrical current?
electroporation
What method of inserting a desired gene into an organism’s DNA involves carrying the DNA into cells by a viral vector that can either be injected straight into the bloostream or deliever by aerosal
transduction (by vector)
How can transduction (by vector) be used in real life or in the future?
nasal spray or oral aerosol, like for asthma
What is a viral vector?
A form of a virus used to deliver genetic material into a cell
What are some example of a viral vector?
adenovirus, liposome
or bacterial plasmid
What is gene therapy?
a new form of medicine, where a healthy copy of a gene is inserted into defective non-germline tissue in a developed or mature plant or animal. Non germline = not passed on to offspring
What is genetic engineering?
the deliberate modification of the characteristics of an organism by manipulating its DNA
What are the five techniques for Recombinant DNA?
T.GS.GT.E/C
1) Transgenesis
2) Gene Sequencing
3) Gene Therapy
4) ELISA
5) CRISPR
What is transgenesis?
introducing DNA from an external source/different organism (exogenous genetic material) into a living organism
What is exogenous?
from an external source or different organism?
What is the point of transgenesis?
so the organism can exhibit a new trait and transfer this trait to its offspring
Why are we even able to put DNA from one organism into another and it works? Why does DNA do the job regardless of who or what it’s in?
because a defined sequence of DNA will code for the same protein in all organisms. So DNA is basically a universal language that can be applied in different organisms in different species
What is the most common form of transgenesis?
gene editing in bacteria
What is the key component needed to gene edit in bacteria and why?
naturally occurring plasmids or circular DNA.
because genes can be easily inserted into the,
All organisms store their genetic code in?
DNA
How is recombinant DNA useful in vaccine research? (with example, mention Hep.B)
recombinant DNA caccines can be produced by selecting a gene that codes for a particular antigen, instead of using the whole DNA of the antigen.
Like with the Hep.B vaccine, the target DNA is inserted into a plasmid that is recombined with a yeast cell. This recombined cell is incubated to let the yeast cells multiply. Then the antigen is extracted and purified as a vaccine.
Vaccine research using recombinant DNA is currently beign used to develop vaccines against…?
malaria and cattle tick
What’s a positive of recombinant DNA vaccines?
- low risk of side effects
- relatively cheap to produce
(so good option for developing countries)
What is the process of using organisms from other animals?
Xenotransplantation
How would xenotransplantation be used with pigs?
transgenic pig organs that have proteins on the surface of their cells to show the cell’s lineage and stage of differentiation, would stop the activation of organ rejection, allowing people on a long waiting lsit for an organ to access viable one
