1. Microorganisms
In biotechnology

2. Microorganisms & history
Throughout history man has been afflicted by disease. The impact of these diseases have changed the course of history:
Malaria- eukaryotic protozoan-Plasmodium. Contributed to the death of King Tut ˜3,000 years ago and the decline of the Roman Empire˜1,500 years ago.
Anthrax- Bacillus anthracus – source of the fifth plague of Egypt mentioned in the Book of Exodus in the Bible.
Plague- Yersinia pestis– source of the Black Death that killed millions of people during the Middle Aged and resulted in the loss of 30-60% of Europe’s population.

3. Model Systems
In the 1870s German Physicist Robert Koch was able to further studies performed by Davaine and Rayer who used Antrhax as a model system in studying livestock diseases that infect humans. Koch discovered that the anthrax disease was caused by anthrax endospores that persist in the soil.
Model systems are useful for gaining knowledge that can be applied to other systems.
In the 1880s French chemist Louis Pasteur used weakened forms of anthrax to immunize sheep. Pastuer used what he learned from anthrax to develop a vaccine against rabies.

4. Koch’s Postulates
Koch & Pasteur are recognized as two founders of microbiological research. Koch used his knowledge from anthrax and TB research to develop postulates to determine if a microorganism is the cause of a disease.
Postulates:
1. The microorganism must be found in abundance in all organisms suffering from the disease but should not be found in healthy organisms.
2. The microorganism must be isolated from a diseased organism and grown in pure culture.
3. The cultured organism should cause disease when introduced into a healthy organism.
4. The microorganism must be re-isolated from the inoculated diseased experimental host and identified as being identical to the original causative agent.
Theses postulates were responsible for the discovery of: diphtheria, pneumonia, meningitis, leprosy, plague and syphilis.

5. Uses of Bacteria in Biotechnology
Food Production
Yogurt- lactic acid fermentation
Lactobacillus delbrueckii subspecies bulgaricus
Protein Production
A protein that is artificially produced in a genetically engineered organism is called a recombinant protein.
To produce a recombinant protein in bacteria, the gene for the protein is cloned and transferred into the bacteria, it is then cultured which means they are placed in conditions that enable them to multiply into billions of bacteria. Once the culture has reached the appropriate density the bacteria are harvested and lysed. The recombinant proteins are then purified from bacteria using chromatography. Uses for recombinant proteins include therapeutic drugs, agriculture and food production.

6. Insulin
Insulin used to treat diabetes is manufactured as a recombinant protein.
Historically purified from pig pancreas.
Patient reactions and expense of purification
1978 Genentech cloned the human insulin gene and genetically engineered E coli bacteria to produce human insulin
Eli Lilly liscenced and sold under the name Humulin in 1982
Humulin is now produced in Eukaryotic cells

7. LAB SKILLS TO DATE Follow laboratory protocols
Make dilutions from concentrated solutions
Use an adjustable volume micropipet
Make molar solutions using a volumetric flask
Use a serological pipet with a pump or aid
Use and calibrate a pH meter
Use a transfer pipet
Label agar plates and broth
Perform calculations C1V1=C2V2
Make LB, LB/AMP, LB/AMP/ARA agar and broth
Wash glassware properly
Use aseptic technique
Make percent solutions
Write an SOP
Write a label for a reagent bottle
Use a spectrophotometer
Use a graduated cylinder
Calculate molarity
Use a balance

8. Upcoming Lab Skills Inoculate an agar plate for a bacterial lawn
Analyze an agarose gel
Streak bacteria to isolate single colonies
Transform bacteria by calcium chloride transformation
Aseptically inoculate a plate with liquid culture
Perform HIC chromatography
Aseptically inoculate a liquid culture from a plate
Perform SDS- polyacrylamide gel electrophoresis
Identify bacteria from cell shape
Stain Polyacrylamide Gels
Prepare agarose gels
Dry polyacrylamide gels
Load an agarose gel
Perform a qualitative ELISA
Perform agarose gel electrophoresis
Use a power supply