BITC1311 Intro to Biotechnology
Chapter 5. Microbial Biotechnology
Name
Thieman & Palladino Guided Notes
1.
Compare bacterial cells with those of eukaryotes with respect to size and cellular structures.
2.
Compare bacterial chromosomes with those of eukaryotes with respect to size and structure.
3.
What are some fungal products that are valuable in the biotechnology industry?
4.
Compare yeast cells with those of bacteria with respect to size, cellular structures and chromosomes.
5. Can you think of a reason that the ability to grow anaerobically can make large-scale growth of yeasts an
advantage?
6.
What are some advantages of the use of Pichia pastoris in large-scale fermentations.
7. Describe some uses by consumers for the following enzymes produced by microbes: cellulases, proteases,
amylase, polymerases.
8. Describe the steps for making bacterial cells competent for transformation, explaining the mechanism by which
each step makes the transformation frequency to improve.
a. Describe how and why transformants must be selected for in selection medium.
b. Describe 4 advantages of electroporation over the calcium chloride method of transformation.
9.
Describe an advantage for the engineering of a fusion protein in an expression vector.
a. What sort of affinity ligand would be used in affinity chromatography of a fusion protein containing a
glutathione-S-transferase tag? How would you remove the purified protein from such a column?
b. How can a tag protein be removed from a fusion protein?
c. What are lux genes, and how are they used as “reporter genes”?
d. What are the substrates of the luciferase enzyme?
e. Describe an application of a lux gene for detecting a tuberculosis infection. Why is this an advantage over
a direct assay for the pathogen?
f. Describe how the yeast two-hybrid system is used to determine what proteins bind to each other. What
does the “bait” protein part of this system fused to? What is the “prey” protein part of this system fused to?
How can you tell when the “prey” has taken the “bait”?
10. What role does rennin play in the production of cheese?
a. What is the traditional source of rennin for the cheese industry?
b. What is the recombinant source of rennin and what is it called?
c. What advantages are there in the substitution of chymosin for rennin?
11. Describe what becomes of the following molecules during the metabolism of glucose by glycolysis: ADP,
ATP, NAD+, HADH, glucose, pyruvate. Is oxygen required to complete this glycolytic metabolism of glucose?
12. Why do anaerobic cells produce alcohol or lactic acid? Why are bubbles of air produced during alcoholic
fermentations such as in the production of beers and champagnes?
13. Describe the process for making yogurt.
14. Describe the causes and therapy for Type I diabetes.
a. What are some problems with the extraction of insulin from pig and cow carcasses?
b. Describe how insulin is produced in pancreatic cells from preproinsulin precursor protein.
c. Describe how recombinant insulin is produced from fusion proteins.
15. Describe how Alexander Fleming discovered penicillin in 1928.
a. Describe the mechanism of action of some common antibiotics that can be used against bacterial
pathogens.
b. How are “inducer” microbes been designed to help solve problems with antibiotic-resistant pathogens?
16. Describe the experiment that was the first deliberate release of a genetically engineered organism into the
environment in 1987.
17. Describe how Edward Jenner developed the first vaccine in 1796.
a. a. What are the diseases targeted by the following vaccines: DPT, MMR, and OPV?
b. b. What are antigens and what types of molecules make up antigens?
c. c. What type of immune cell produces antibodies?
d. d. During a primary response to an antigen, what role do plasma and memory cells play?
e. e. What roles do antibodies play in eradicating pathogens? What other immune cell and immune system
proteins play a role in this process?
f. f.
What is the difference between a subunit vaccine and a whole cell vaccine? What are some
advantages of each?
g. g. What is the difference between an attenuated vaccine and an inactivated vaccine? What are some
advantages of each?
h. h. Describe how recombinant DNA can be used to make subunit vaccines.
18. What are 4 basic components of a virus such as HIV?
a. Describe the processes of the HIV life cycle, paying attention to key components involved in each: 1)
binding 2) Infection 3) reverse transcription 4) genome integration 5) genome replication 6) protein
synthesis 7) protein cleavage 8) viral particles assembled
b. Give some examples of viral infections that are widespread in the human population.
19. Describe how sequencing projects can lead to better diagnostics of and ways to treat infectious conditions.
a. What proportion of bacterial genomes sequenced to date code for genes of unknown function?
b. What proportion of bacterial genomes sequenced to date code for genes that are unique to the bacterial
genome being sequenced?
c. How is “annotation” used to assign genetic function to sequencing data?
20. Describe some general strategies by which antiviral drugs work: what are their therapeutic targets?
21. Before the advent of DNA technology, microbiologists relied on growth on different growth media and
biochemical tests to identify strains of disease-causing bacteria.
a. What are some molecular biology techniques that are used to identify strains of bacteria for diagnosis and
prevention of disease?
b. What is PulseNet, and how is it used to prevent disease outbreaks?
22. Describe some situations in which bioweapons had been used early in the history of Western civilization.
a. What are some proposed avenues for spreading a bioweapon in a human population by bioterrorists?
b. What are some other possible means other than spread of human diseases that bioterrorists could attack?
c. Describe how DNA technology can be used to detect a bioweapon and track its origin.