1. RECOMBINANT DNA GENETIC ENGINEERING PART I
BEGINS

2. The Transformers, more than meet the eyes
The Transformers, more than meet the eyes. The Transformers, robots in disguise.

3. What a blast from the past.
Hey kids, I’m here to present the concepts of recombinant DNA. Now you might wonder why I am presenting. Well, recombinant DNA involves taking regular bacteria and TRANSFORMING them into new types of bacteria that express different and useful genetics to make things such as hormones for humans. It’s a little like me helping humans by transforming myself into my super-strong robot configuration.
What a blast from the past.

4. Here I am in my unassuming form
Here I am in my unassuming form. I can cruise the streets unnoticed because I’m just a car, with extremely cool looks by the way.
This bacteria is also a common bacteria. These bacteria are nothing special, but they can be transformed! Transformation in bacteria is nothing new nor is it something that was invented by humans. Transformation was discovered by Griffith in his 1928 experiment The Avery-MacLeod-McCarty experiment characterized the DNA as being responsible for the transformation. .
Dr. Avery here. We should have got the Nobel prize! The Nobel foundation expressed public regret for failing to award it to us.

5. Hey give me back my pancreas!
In the early 1970’s Stanley Cohen (plasmid guy) and Herbert Boyer (restriction endonuclease guy) combined to form the early recombinant DNA work. In 1976 Boyer cofounded Genetech, a premier genetic engineering company on the Stock Exchange.
The challenge for this company was to develop recombinant techniques so that bacteria could express foreign genes and manufacture products that they normally would not. These products could be therapeutic chemicals such as human growth hormone or insulin.
Prior to this, insulin was only available on a small scale. It was not as effective because it had to be extracted from animal pancreas and purified. This process was very difficult and so insulin was expensive to purchase. Recombinant technology could make this hormone on a massive scale, yet at a much greater reduction in price!
Hey give me back my pancreas!

6. Cadavers , those are dead bodies! I like bacteria better.
Let’s get rich and make life better for all the diabetics at the same time.
If we use recombinant DNA to get bacteria to make insulin, then we can produce massive amounts reducing the price to the public and making us a hefty profit in the process. We just have to take the theory and apply it to an actual manufacturing process.
Human growth hormone was even more difficult to isolate and extract. It had to be taken from human cadaver pituitaries and thus presented the risk of infections from cadaver to recipient. Cost to the public was so expensive, it was virtually unaffordable for people to seek therapy.
Recombinant technology changed all that posing no risk of infection because bacteria manufacture the gene. It also tremendously reduced the cost, making it readily available to the public.
Cadavers , those are dead bodies! I like bacteria better.

7. Prions evil like me but not cells!
I am a MAD COW! Don’t eat me because you will get the prion and you will get my incurable disease!
DID YOU KNOW?
The concern with cadavers transferring infection from their pituitary gene is from a disease called Creutzfeldt–Jakob disease. This disease is better known as MAD COW DISEASE. It has no cure and is caused by a prion. These are particles that are neither viruses or bacteria. They are simply proteins!
Although CJD is the most common human prion disease, it is rare. Incidence-- 1 in a million people per year. It usually affects people aged 45–75 but teenagers are a recent addition. Death within a few months
This nasty little protein called a prion causes other proteins to bend into the wrong shape, become insoluble, useless and worse of all, incapable of being digested. The accumulation of these proteins causes plaques and holes to develop in the brain cells shown at the side.
Prions evil like me but not cells!

8. AIDS associated wasting disease
You might ask, “Why is there a demand for the growth hormone, somatotropin?”. After all, profit in pharmaceuticals is what it is all about and profit means we need more people buying this biotechnology product.
OUR CUSTOMERS -- People seeking treatment for:
Dwarfism
Turner’s syndrome
AIDS associated wasting disease
Shhhh ----Miracle muscle and youth rejuvenating agent for rich people athletes and movie stars (unproven). But you didn’t hear it from me.

9. In this process of recombinant DNA, we need to be able to control the bacteria. We need knowledge of an operon gene to be able to control when the bacteria would make the growth hormone (turn it on or off).
Remember my lesson on the LAC OPERON? Now you should be able to apply that knowledge to make it useful for this biotechnology application
Utilizing a plasmid with the new growth hormone gene inserted, the plasmid was then inserted into the bacteria. Due to research in plasmid mapping, endonucleases could be used to cut the bacterial genome in a precise location, allowing the plasmid to insert after the lac promoter. This put the growth hormone under the control system of the lac operon.
The bacteria of choice -- E. coli cells. Normally the gene is activated by the presence of the inducer, LACTOSE.
The chemical isopropyl thiogalactoside (IPTG) is an inducer of the Lac operon in bacterial cells. Therefore, in the presence of IPTG, bacteria are induced to produce somatropin. In the absence of IPTG, somatropin is not produced, allowing control of the hormone produced.

10. Did you notice they called the drug somatropin
Did you notice they called the drug somatropin. It’s artificial somatotropin and goes by a different name.
Now lets get to the “nitty gritty” process of transforming bacteria!! This is what we want:
Transformed into
Note: this bacteria only makes somatropin when we put it in IPTG.
Transformed into
E. coli
E coli making somatropin

11. Please continue with the next session
RECOMBINANT DNA
Please continue with the next session