MICRB320 Lecture Notes - Lecture 7: Dna Extraction, Dna Replication, Molecular Ecology

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orangecheetah563

22 Oct 2015

School

University of Alberta

Department

Microbiology (Biological Sciences)

Course

MICRB320

Professor

Rebecca Case

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Document Summary

Class presentations: first presentation is tuesday 11 february, paper is given one week before presentation (tues 4 feb, slides are due by 4pm the day before your presentation. Methods, what they did conclusion - what we know, what we found. At least one (1) slide with a figure from the paper. At least one (1) slide summarizing the findings of the paper. At least one (1) slide as a critic of the paper. At least one (1) conclusion slide: presentation will begin at 2. 55pm, presentation is 10 min (5 marks) Aim for 8 min: lead discussion for 10 min (5 marks) Allow us to see cells in an environment and to quantify them. Without culturing - counts for every cell. Soil - not accurate to count for cfu. Compare the sequences to allow us to identify bacteria. Within their habitat: cells could be counted allowing experiments to become quantitative (not qualitative, identification based on 20-40 bp probes.

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GENETICS QUESTION: For this experiment we focused on forward genetics: First, his- mutants were identified and then complementation analysis was used in order to determine which of the 7 his genes was mutated. ( The exact data can be seen at the bottom). In this experiment we screened for his- mutants that were generated via a random mutation process resulting from errors in DNA replication or induced by EMS aka Ethyl methanesulfonate. We compared a mutagenized culture that was treated with EMS and an un-mutagenized culture that was not treated with EMS. However, both cultures went trhough an enrichment process to increase the frequency of his- mutants.

Point mutations involve the substitution of one type of base pair for another. If the substitution replaces a purine with a purine (or a pyrimidine with a pyrimidine), it is called a transition. If the substitution replaces a purine with a pyrimidine (or a pyrimidine with a purine), it is called a transversion. Many mutagenic chemicals cause this type of mutation by alkylation of a nucleotide base. The chemically altered base can then be misread during replication or repair. For example the mutagen, ethyl methanesulfonate (EMS), adds an ethyl (-CH2-CH3) group to both guanine or thymine bases causing the modified guanine to pair with thymine and the modified thymine to pair with guanine.

1. Why might one gene be more susceptible to EMS mutagenesis than another gene?

2. Why might we have expected to find a higher frequency of polar mutants (his4A^-B^-C^-) in the unmutagenized culture compared to the mutagenized culture?

Table 2. Experiment 1B class data summary (number of his^-mutants by section) Unmutagenized
Section	1	2	3	*4ABC*	4A	4B	4C	5	6	7	*Total his^-* mutants**
Tuesday	7	1	5	4	2	1	9	2	3	1	35
Wednesday	2	1	14	0	4	2	2	4	8	2	39
Total	9	2	19	4	6	3	11	6	11	3	74
% each mutant	12.1	2.7	25.7	5.4	8.1	4.1	14.9	8.1	14.9	4.1	~100%

Mutagenized
Section	1	2	3	4ABC	4A	4B	4C	5	6	7	*Total his^-* mutants**
Tuesday	2	5	4	2	6	5	5	0	4	1	34
Wednesday	9	1	15	0	9	1	2	2	9	4	52
Total	11	6	19	2	15	6	7	2	13	5	86
% each mutant	12.8	7.0	22.1	2.3	17.4	7.0	8.1	2.3	15.1	5.8	~100%

Create a new dictionary called prices_stocks using {} format like the example below.

prices_stocks={}

Put these values in your prices_stocks dictionary:

"banana": [4,5]
"apple": [2,2]
"orange": [1.5,2.5]
"pear": [3,4]

So the key of dictionary is name of item and value is a list (first element of that list is stock and second element is relevant price)

Loop through each key in prices_stocks, for each key, print out the key along with its price and stock information. Finally calculate how much you earn if you sold all of your products. Print the answer in the following format:

Part-1:

Item: banana Stock: 4 Price: 5

Item: apple Stock: 2 Price: 2

Item: orange Stock: 1.5 Price: 2.5

Item: pear Stock: 3 Price: 4

If you sold all of your items you earn 39.75

azureraccoon643

QUESTION 30

The function of centrosome is:

	1.	Microtubule initiation and organization in animal cells
	2.	Organization of thin filaments
	3.	Depolymerization of microtubules
	4.	Organization of intermediate filaments

2.5 points

QUESTION 31

The destinations of proteins synthesized on "free" ribomsomes include:

	1.	Endoplasmic reticulum
	2.	Nucleus
	3.	Mitochondria
	4.	Lysosomes

2.5 points

QUESTION 32

Microtubules can be assembled and disassembled quickly in response to the cellular changes is because:

	1.	Cells synthesize tubulin proteins very fast.
	2.	Cells can turn other cytoskeletal filaments into microtubules
	3.	Microtubules have an instable plus end and microtubules are probably organized by weak non-covalent interactions
	4.	The motor proteins of microtubules can organize the microtubules in response to the cellular changes

2.5 points

QUESTION 33

Both signal recognition particle and a translocon in rough endoplasmic reticulum membrane can recognize and bind to the signal sequence of a secretory protein, so they play the exact same role during protein co-translational translocation.

True

False

2.5 points

QUESTION 34

Rough endoplasmic reticulum is so named is because:

	1.	It has many proteins associated with its surface.
	2.	It has ribosomes associated with its cytosolic surface.
	3.	It is associated with nuclear membrane.
	4.	It has vesicles budding off its surface.

2.5 points

QUESTION 35

The membrane vesicles that transport materials between membrane compartments have a protein coat. The protein coat has similar functions in different vesicles. These functions include:

	1.	Help the enclosed material fold correctly.
	2.	Help degrade the protein in the vesicles
	3.	Maintain the internal pH of the vesicles
	4.	Function as mechanical device to form a vesicle and select the components to be carried by a vesicle.

2.5 points

QUESTION 36

Lysosomes are important for:

	1.	Receptor-mediated endocytosis.
	2.	Breaking down foreign particles brought into the cells by endosomes
	3.	Regulating the process of organelle turnover.
	4.	Protein trafficking between membrane compartments

2.5 points

QUESTION 37

Which of the following is correct about membrane biosynthesis?

	1.	Membranes are synthesized in Golgi Complex.
	2.	Membranes arise from preexisting endoplasmic reticulum membranes.
	3.	Membranes are synthesized on ribosomes.
	4.	Membranes of all organelles have the same composition of proteins and lipids, since they all originate from endoplasmic reticulum.

2.5 points

QUESTION 38

During skeletal muscle contractions, the shortening of the muscle fibers is because of:

		The shortening of the thin and thick myofilaments.
		The shortening of the cytoskeletal filaments.
		The kinesin proteins moving over microtubules.
		The actin and myosin filaments sliding over each other.

2.5 points

QUESTION 39

The assembly and disassembly of microtubules is regulated by rate of growth and shrinkage of the plus end.

True

False

2.5 points

QUESTION 40

The functions of Golgi complex include:

	1.	O-linked oligosaccharides and most of a cellâs complex polysaccharides are assembled in Golgi complex
	2.	Cis Golgi complex sorts proteins
	3.	Trans Golgi complex segregates proteins in different types of vesicles for dilevery.
	4.	N-linked oligosaccharides are further processed in Golgi complex

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MICRB320 Lecture Notes - Lecture 7: Dna Extraction, Dna Replication, Molecular Ecology

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