BIOL 3581 Lecture 11: Biotech Lecture 11 (Nov 28)

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orangehorse801

7 Feb 2017

School

University of Windsor

Department

Biological Sciences

Course

BIOL 3581

Professor

Vacratis

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Document Summary

Idea of having ability to analyze known protein complexes, but especially unknown ones. With that knowledge, you can advance knowledge of existing protein if you find new types of associations (maybe in different cell type or different conditions) Or you can use it for an unknown protein that is poorly characterized. In a fairly reproducible, feasible, and in a short amount of time (6 months to a year is considered short) This experiment can be done in a couple weeks. You can learn something monumental about an unknown protein. Sample preparation should be done by cell biologists. That type of analysis is under the general umbrella of proteomic research. Proteomics can be defined in different ways by different people. Two main areas: identify the components of a particular proteome, see how levels change. Proteomics can be used as a database. Any of these (cid:498)compartments(cid:499) that people study using mass spec, doing it at a high.

Related Questions

You have done a genetic screen to identify genes controlling wings in an insect. Some of the mutants have no wings, some have short wings and others have larger than normal wings.

1a. (9 points) What type of mutation (e.g., amorph, hyopmorph, etc.) is most likely responsible for each class of phenotypes? Explain briefly. State whether you expect each to be recessive or dominant.

No wings:

Short wings:

Large wings:

Mutation	1	2	3	4	5	6	7	8	9	10	11	12
1	-	+	+	+	-	+	+	+	+	+	+	+
2		-	+	+	+	-	+	-	+	-	+	+
3			-	-	+	+	+	+	+	+	+	+
4				-	+	+	+	+	+	+	+	+
5					-	+	+	+	+	+	+	+
6						-	+	-	+	-	+	+
7							-	+	+	+	-	-
8								-	+	-	+	+
9									-	+	+	+
10										-	+	+
11											-	-
12												-

1b. (2 points) How many genes causing the small wing phenotypes are revealed by this analysis? ___________

1c. (6 points) Which mutants belong to the same complementation groups?

1d. (3 points) Cloning of the gene causing the large wings phenotype reveals that is a growth regulator, which is a ubiquitous protein present in many different cell types. Suggest an hypothesis to explain why only the wings were large but not other tissues.

coffeewasp36

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 (ÎlepA , 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

ochregerbil875

__________ are fairly small organelles that provide a safe place within the cell to carry out certain biochemical reactions that generate harmful, highly reactive oxygen species. These chemicals are both generated and broken down in the same location.

Lysosomes

Endosomes

Peroxisomes

Nucleosomes

Biologists cannot possibly study all living species. Instead, they try to understand cell behavior by studying a select subset of them. Which of the following characteristics are useful in an organism chosen for use as a model in laboratory studies?

rapid rate of reproduction

all of the above

ability to grow under controlled conditions

amenability to genetic manipulation

By definition, prokaryotic cells do not possess __________.

ribosomes.

a nucleus.

membrane bilayers.

replication machinery.

Cell biologists employ targeted fluorescent dyes or modified fluorescent proteins in both standard fluorescence microscopy and confocal microscopy to observe specific details in the cell. Even though fluorescence permits better visualization, the resolving power is essentially the same as that of a standard light microscope because the resolving power of a microscope is limited by the __________ of light.

intensity

absorption

wavelength

filtering

Choose the phrase that best completes this sentence: Microtubules ____________ and are required to pull duplicated chromosomes to opposite poles of dividing cells.

generate contractile forces

can rapidly reorganize

are found in especially large numbers in muscle cells

are intermediate in thickness

Despite the differences between eukaryotic and prokaryotic cells, prokaryotes have proteins that are distantly related to eukaryotic actin filaments and microtubules. What is likely to be the most ancient function of the cytoskeleton?

cell division

cell motility

vesicle transport

membrane support

DNA and RNA are different types of nucleic acid polymer. Which of the following is true of DNA but not true of RNA?

It contains thymine.

It has 5'-to-3' directionality.

It is single-stranded.

It contains uracil.

Double covalent bonds are both shorter and stronger than single covalent bonds, but they also limit the geometry of the molecule because they ____________________.

prevent additional bonds from being formed with the bonded atoms.

limit the rotation of the bonded atoms.

create a new arrangement of electron shells.

change the reactivity of the bonded atoms.

Drosophila melanogaster is a/an __________. This type of animal is the most abundant of all animal species, making it an appropriate choice as an experimental model.

mammal

amphibian

bird

insect

10.

If the isotope 32S has 16 protons and 16 neutrons, how many protons, neutrons, and electrons will the isotope 35S have, respectively?

16, 19, 17

16, 19, 15

16, 19, 16

16, 20, 15

11.

Living systems are incredibly diverse in size, shape, environment, and behavior. It is estimated that there are between 10 million and 100 million different species. Despite this wide variety of organisms, it remains difficult to define what it means to say something is alive. Which of the following can be described as the smallest living unit?

cell

organelle

protein

DNA

12.

Macromolecules in the cell can often interact transiently as a result of noncovalent interactions. These weak interactions also produce stable, highly specific interactions between molecules. Which of the factors below is the most significant in determining whether the interaction will be transient or stable?

the concentration of each molecule

the size of each molecule

surface complementarity between molecules

the rate of synthesis

13.

Many of the mechanisms that cells use for maintenance and reproduction were first studied at the molecular level in bacteria. Which bacterial species had a central role in advancing the field of molecular biology?

E. coli

S. pombe

C. elegans

D. melanogaster

14.

Many types of cells have stores of lipids in their cytoplasm, usually seen as fat droplets. What is the lipid most commonly found in these droplets?

cholesterol

isoprene

triacylglycerol

palmitic acid

15.

Mitochondria contain their own genome, are able to duplicate, and actually divide on a different time line from the rest of the cell. Nevertheless, mitochondria cannot function for long when isolated from the cell because they are __________________.

anaerobes.

viruses.

parasites.

endosymbionts.

16.

Mitochondria perform cellular respiration, a process that uses oxygen, generates carbon dioxide, and produces chemical energy for the cell. Which answer below indicates a correct pairing of material âburnedâ and the form of energy produced during cellular respiration?

17.

Polar covalent bonds are formed when the electrons in the bond are not shared equally between the two nuclei. Which one of these molecules contains polar bonds?

BIOL 3581 Lecture 11: Biotech Lecture 11 (Nov 28)

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