BIOL308 Study Guide - Islamic Republic News Agency, Wobble Base Pair, Transfer Rna

CRISPR

Previous genetic editing mechanisms

What are the Talen and Zinc finger approach?

Why were they difficult?

CRISPR in nature

How was it found? What does the second R stand for in CRISPR?

How does it work as an immune system?

CRISPS as a gene editing tool

Give 3 names of people who found it. Who found it first? Why is it controversial about the patent?

Explain role of

CAS9

gRNA sequence

Donor DNA

Homologous recombination

NEHJ

How can CRISPR be used?

What are the considerations when using it in humans?

What is a gene drive?

How is it different than CRISPR

What is a daisy drive?

2. Explain the structural role of the four different categories of biomolecules within a cell.

3. Explain how water, a steroid hormone, and a monosaccharide are transported into a cell down their concentration gradient and explain why the different molecules are transported in different ways.

4. Explain which forms of transport require energy and why, and how ATP facilitates energy-dependent transport.

5. All of the different types of biomolecules can serve as energy sources. Explain which biomolecules are catabolized in which metabolic pathways.

6. Whether or not the cellular machinery is in place for particular catabolic reactions to go forward depends on which genes are being expressed. Explain the different points, in a eukaryotic cell, at which gene expression might be regulated, and how.

7. The primary tasks of any organism are survival and reproduction. Acquiring biomolecules and catabolizing them to generate energy is a major factor in survival. For reproduction to occur, DNA must first be replicated so that it can be passed on. Describe the process of DNA replication.

8. If you wanted to replicate DNA artificially in large quantities, how would you go about it?

9. Once DNA has been replicated, what is the sequence of events in the cell cycle and in cell division that produce the gametes for sexual reproduction? Where does this event fit, along with other cell cycles, in the life cycle of the organism?

10. A number of physical traits in humans are controlled by a simple dominant/recessive relationship between two alleles; here are some examples (dominant/recessive):

Widow’s peak/no widow’s peak

Free earlobes/attached earlobes

Cleft chin/no cleft chin

Dimples/no dimples

Freckles/no freckles

Naturally curly hair (really curly, not wavy)/straight hair

Make a pedigree of your immediate family indicating the possible genotype of each family member (for the six genes listed above) based on their phenotype. If you are adopted, indicate the possible genotypes and phenotypes of your parents. You may make this part up.

Please I need help answering those questions i always lose bunch of points i'm not doing it correctly.

Use information from Nature Reviews Microbiology to answer the following question.What is the role of EF-G and EF4 in protein synthesis? Under what circumstances does EF4 participate in translation and what is its role?

from this reading The Highly Conserved LepA Is a Ribosomal Elongation Factor that Back-Translocates the Ribosome Qin Y,Polacek N, Vesper O, Staub E, Einfeldt E, Wilson D.N, and Nierhaus N.H. Cell 127: 721-733 (2006).

Q2. In which species is Lep A found and in which ones is it absent? Is the result expected based on your knowledge of the endosimbiosis theory?

Q3.

3a What is the percentage of identity between LepA and its ortholog genes?

3b. (5pts) Indicate what domains are conserved between Ecoli EFG and LepA, and which ones are not.

3c. What do grey and black boxes in the sequence alignment indicate?

Q4. Analysis of in vivo experiments shown in Figure 3. Only analyze graph on the left side of the figure.

4a. What is the difference between the bacteria whose growth are shown by the blue and the green curves? What can be concluded about the effect of EFG on bacterial growth?

4b. Which protein is expressed by bacteria whose growth is shown by the yellow curve? Does the expression of this protein affect bacterial growth? Compare the yellow and blue curves. How can this result be explained?

4c. Explain why the addition of IPTG results in lower bacterial growth. Compare the yellow and red curves?

4d. Why is the green curve an important control of the experiment? The green curve.

The next questions are based on the reading of the article Elongation factor 4 (EF4/LepA) accelerates protein synthesis at increased Mg2+ concentrations.

Elongation factor 4 (EF4/LepA) accelerates protein synthesis at increased Mg2+ concentrations Pech M, Karim Z, Yamamoto H, Kitakawa H, Qin Y, and Nierhaus K. PNAS 108: 3199-3203(2011)

Studies done in E. coli using a knockout mutant of the lepA gene (ΔlepA , lepA gene has been deleted) did not show any phenotype when grown in rich LB medium. This result was unexpected due to the high level of conservation of EF-4 among species.

Q5. Equal amounts of wild type and mutant cells (lacking the lepA gene) were mixed and co-cultured for 24h. The population distribution was determined every 4 hours. Figure 1 indicates changes in the % of mutant cells over several generations.

Q5a. What is the question that researchers addressed in the experiments shown in Figure 1?

Q5b. Which curve represents the control of the experiment? Do the two strains grow at similar rate? How do you know?

Q5c. Do high salt concentration and/or low temperature have any effect on the growth of mutant cells?

Q5d. What can you conclude about the role of LepA on bacteria growth? When is this protein important?

Q6.

6a. (6pts) Where in the cell is EF4 located? Did you expect that this would be the EF4 location? Why or why not?

What is the question that the researchers want to address in the experiment shown in Figure 2?

6c. Analyze the green and red bars graph shown in Figure 2. What technique was used to detect EF4? Is there any change in EF4 localization?

Q7. Summarize the main points of the model shown in Figure 4