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Unit 11 A4 Learning aim D: Explore basic DNA techniques and the use of genetic engineering technologies
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1.Explain and analyse the stages involved in medicine for the development of insulin.Before the discovery of insulin, diabetes was known to be life-threatening, however, due to the help of
science, diabetes is now manageable across the world. Insulin discovery started from 1700-1923. By Dr,
John Rollo a British physician, who had the idea of diabetics of a low-carbohydrate diet high in fat & animal
protein and then it was Dr Fedrick M.Allen’s idea of a near-starvation diet, however, what Dr John Rollo did
was not that helpful, as Dr Fedrick M. Allen’s decided that by doing so led to diabetic patients in getting
severely malnourished. Then in 1920, Dr Frederick Banting wanted to make a pancreatic extract for diabetes,
which will have some of the anti-diabetic qualities. As so in 1921, Banting along with medical student
Charles Best both of whom decided to make a pancreatic extract. They used an animal which was a dog, for
which they surgically ligated (prevent bleeding during surgery), as this stopped the flow of nourishment for
the pancreas degenerated.
As so, by removing the pancreas, slicing it up, and then freeze the pieces in a solution of water & salt. But
then managed to ground up and filter the frozen pieces. And extract something that was insulin. So then in
1922 after a year, insulin was then tested on Leonard Thompson, that a 14-year-old patient who had diabetes,
the patient was lying dead, but amazingly Professor. J. Macleod managed to save him. Banting and Macleod
were awarded the Noble Prize in Physiology in 1923. Overall, insulin did not cure diabetes but was a
treatment that was a stepping-stone in science. When you eat, glucose will be used in the body for energy.
It’s created by many of the carbohydrates like pasta, bread, biscuits, etc. Glucose will then either be used or
stored in your cells and this is where insulin plays a part. Insulin will help keep the glucose in your blood
that is within a normal range. It helps to manage the regulation of the metabolism of carbohydrates, proteins,
and fats as it helps in promoting the absorption of glucose that’s needed for the body as so it helps to
promote the absorption of glucose from the blood into that of the liver, skeletal, and fat cells. After all, it
takes glucose out of the bloodstream which glucose will be taken out of the bloodstream and moving into the
cells throughout the body and what’s amazing about insulin is that the cells will use the glucose for energy
and store the excess in the liver, muscles, and fat tissue cells. As so too much or too little glucose will then
cause dangerous health problems, conditions like diabetes will develop, it can go on to get heart, kidney, eye,
and blood vessel problems. As type 1 diabetes, in the pancreas, it takes longer to be able to produce insulin
and in type 2 diabetes that the pancreas will be produced insulin, as the cells of the body is unable to make
good use of the insulin. So, what happens if the diabetes is unmanaged? This allows the glucose to then build
up in the blood and will not be able to distribute to the cells. As such this can then wreak havoc with every
part of your body. The negative feedback loop is that as the blood sugar rises in the blood, insulin will then
sends signals to the organs such as the liver, muscles, and the other cells that store the excess glucose that’s
the negative part of it as it's stored it will be stored as the body fat. The body will try to store other parts that
will be stored as glycogen in the liver and the muscles as so if the blood glucose level is too low, which
cause the pancreas to release the hormone glucagon as its distributed around. (Anon., 2019), (StudyMoose,
2020).
How the isolation of two genes is obtained? The amino acid sequence if known can help in knowing, the
DNA code which the DNA can help by putting together the correct order of the nucleotides, however, the
proteins can be very large and would take a long time for it to know the sequence. Then you need to isolate
the mRNA for the desired gene and so it makes a single copy of the complementary DNA. Afterwards, the
DNA uses the enzyme reverse transcriptase and then another copy will be done to add DNA polymerase and
so this can be a doubled stranded length of DNA is made. Then finally to isolate the gene from the entire
genome as the DNA first needs to be cut into fragments so the one that contains the desired gene must be
identified because the enzymes used to cut are called restriction enzymes. For step 1: the bacterial plasmids
can be used to put the insulin gene and the DNA from humans that can include the insulin gene. The second
step is that once it’s isolated the bacterial plasmid and the human DNA will be cut as so to remove the DNA
to get rid of the unwanted DNA. Then restriction enzymes will be used as the same enzyme will be used for
both of the DNA, so the results are of sticky ends.
Step 1: Step 2:
Updated, WHC: June 2017
, Moreover, the DNA fragments will be mixed and cut with plasmids, then will be placed in a test tube, the
sticky ends of the plasmid of the DNA will be formed and the DNA fragments will be joined together by the
formation of hydrogen bonds, and finally using DNA ligase to seal properly. The DNA ligase is that of the
enzyme that will catalyse the joining of sugar and phosphate groups in a strand of the DNA. Now it's
complete.
Step 4: This will require the recombinant plasmid with the bacteria, as so the use of heat shock
transformation will be used and this will require these conditions for these calcium ions to have a high
temperature and this process is explained through this diagram so the normal bacterium will have the
competent cell to have the plasmid bound to the exterior of the cell here in this pink diagram and then as its
left for 2 minutes at 420C and then the plasmid for this transportation into the cell and finally it will form a
transformed cell. Furthermore, this step is essential for DNA cloning as it occurs after restriction digest and
ligation as so it transfers the newly made plasmids and the transformation stage the bacteria will be selected
on the antibiotic plates and this will allow the bacteria that are antibiotic resistant as so each will form a
colony because bacteria colonies with the right plasmids would be grown to make the large cultures of the
identical bacteria that are used to produce plasmid.
Then the culture and purification of the insulin protein, the bacterium individually with have recombinant
plasmid that will produce and as such it will allow isolating the bacterial colonies that gene this is shown in
the diagram below of the process of it. As such bacterial host strain like that of E.coil (Escherichia coli 20),
is amazing as this can provide better and greater efficiency of the new pIBAINS expression, and will enable
to high yield of the possibility of the recombinant human insulin per litre of this amazing engineering of the
development of insulin, as so the plasmid does not integrate into the genome and that remains for it to be
stable and last for a very long time without the need of having an antibiotic selection of it as well as this
process of cell culture which will have through the inclusion of things like that of body isolation and the
dissolution to the protein purification, as such this process can in being a hypoglycemic drug (lower than
what should be within range), as so as many people from different religions are opting for a better way to use
the insulin that is made in a more friendly/vegetarian way (Science Direct, 2019).
Updated, WHC: June 2017
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