Lecture 3 (covers videos A, B, & C) Abbreviations:
Ex. = for example,
Tuesday, January 19, 2021 6:23 PM b/w = between
b/c = because
w/ = with
v. = versus
A. Lab Problem: future direction (follow-up expt) for the biofuels paper? INC = increase
○ Must be related to the results of the paper and be specific(research questions,
steps). C. Analyzing the Biofuel Paper
○ Do NOT do the same procedure w/ another organism! • Main question: is it possible to generate yeast that show INCed tolerance to glucose and
• Antibiotics: ethanol levels?
○ Broad v. narrow(works against certain bacteria) spectrum • Chose to modify TFs (transcription factors) b/c the alterations would affect many genes. ->
○ Static(prevent bacterial growth) v. cidal(kill the bacteria) Better chance at producing an ethanol- and glucose- tolerant mutant.
○ Antibiotics shut down specific pathways in the bacterial target cell: cell wall, protein • Used PCR mutagenesis to modify TFs, and added mutants to WT yeast. Evaluated who
synthesis, nucleic acid, metabolic pathway inhibitors. would survive in an ethanol-glucose rich dish.
○ Fungal genes are much more similar to ours than bacteria. -> Much harder to • Fig. 1B: evaluates survival of mutated TFs in high glucose environment.
target fungal genes specifically/much easier to get cross-reactivity with antifungal • OD: Optical Density. Measures of bacterial/yeast growth.
medicine. • Fold Improvement: how much better is the experimental group growth compared to the
○ Significant INC in the use of antibiotics since 90s -> INC in antibiotic resistance. control group(i.e. WT)?
• Bacteria become resistant through: ○ Measures the OD of SPT15-300 over the WT and the OD of TAF25 over the WT.
○ Spontaneous Mutation: promoter mutations(i.e. overexpression of ribosomal • At 20g/L glucose, the spt15-300 mutant is at 7.8 fold improvement. -> SPT15-300 OD /
proteins), ORF mutation(Open Reading Frame, i.e. structure of ribosomal protein is WT OD = 7.8regular
altered). MODIFYING EXISTING GENOME! • Hypothesis: TPS1 promoter will have an overall higher ethanol production than the SPT15
○ Resistance plasmids that allow acquisition of new genes. Proteins that and PDC mutant strain. PDC promoter will increase in ethanol production as glucose
breakdown/modify antibiotics, drug pumps, new versions of target protein w/ levels increase.
different structure. ADDING SOMETHING NEW!
LECTURE PRACTICE PROBLEMS
Ex. = for example,
Tuesday, January 19, 2021 6:23 PM b/w = between
b/c = because
w/ = with
v. = versus
A. Lab Problem: future direction (follow-up expt) for the biofuels paper? INC = increase
○ Must be related to the results of the paper and be specific(research questions,
steps). C. Analyzing the Biofuel Paper
○ Do NOT do the same procedure w/ another organism! • Main question: is it possible to generate yeast that show INCed tolerance to glucose and
• Antibiotics: ethanol levels?
○ Broad v. narrow(works against certain bacteria) spectrum • Chose to modify TFs (transcription factors) b/c the alterations would affect many genes. ->
○ Static(prevent bacterial growth) v. cidal(kill the bacteria) Better chance at producing an ethanol- and glucose- tolerant mutant.
○ Antibiotics shut down specific pathways in the bacterial target cell: cell wall, protein • Used PCR mutagenesis to modify TFs, and added mutants to WT yeast. Evaluated who
synthesis, nucleic acid, metabolic pathway inhibitors. would survive in an ethanol-glucose rich dish.
○ Fungal genes are much more similar to ours than bacteria. -> Much harder to • Fig. 1B: evaluates survival of mutated TFs in high glucose environment.
target fungal genes specifically/much easier to get cross-reactivity with antifungal • OD: Optical Density. Measures of bacterial/yeast growth.
medicine. • Fold Improvement: how much better is the experimental group growth compared to the
○ Significant INC in the use of antibiotics since 90s -> INC in antibiotic resistance. control group(i.e. WT)?
• Bacteria become resistant through: ○ Measures the OD of SPT15-300 over the WT and the OD of TAF25 over the WT.
○ Spontaneous Mutation: promoter mutations(i.e. overexpression of ribosomal • At 20g/L glucose, the spt15-300 mutant is at 7.8 fold improvement. -> SPT15-300 OD /
proteins), ORF mutation(Open Reading Frame, i.e. structure of ribosomal protein is WT OD = 7.8regular
altered). MODIFYING EXISTING GENOME! • Hypothesis: TPS1 promoter will have an overall higher ethanol production than the SPT15
○ Resistance plasmids that allow acquisition of new genes. Proteins that and PDC mutant strain. PDC promoter will increase in ethanol production as glucose
breakdown/modify antibiotics, drug pumps, new versions of target protein w/ levels increase.
different structure. ADDING SOMETHING NEW!
LECTURE PRACTICE PROBLEMS
