BIO 1140 Chapter Notes - Chapter 12.3: Prophase, Receptor Tyrosine Kinase, Growth Medium

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plumferret19

20 Apr 2018

School

University of Ottawa

Department

Biology

Course

BIO 1140

Professor

Caroline Petit- Turcotte

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Document Summary

12. 3 the eukaryotic cell cycle is regulated by a molecular. How often a cell divides depends on the cell. Human skin cells divide often, liver cells only when needed. 1st cell: this cell was in the s phase. 2nd cell: this cell was in the g1 phase. 1st: this cell was in the m phase. 2nd: this cell was in the g1 phase. In experiment 1, the cell in the g1 phase, when fused with the cell in the s phase, quickly changed to the s phase as well. This is also seen in experiment 2 when both cells fused, the cell in g1 phase quickly changed to the m phase. From this experiment scientist concluded that there are molecules in the cytoplasm during the s or m phase the controls the progression to those phases. Events in the cell are controlled by a cell cycle control system. The cell cycle is regulated by both internal and external factors.

Related Questions

1)What experiment was performed that allowed for scientists to finally agree that DNA is the genetic material within living organisms?

a.Understand the materials and scientific techniques used to come to this result.

2)What group of experiments led to the discovery of the mechanism used in DNA replication?

a.Understand the materials and scientific techniques used to come to this result.

i.What is a cesium chloride gradient?

ii.Where do 14N and 15N respectively fall along this gradient?

b.The results revealed that DNA replicates using which of the three proposed models?

i. Describe the mechanism of each model.

3)What is the role of the following proteins in DNA replication?

a.Helicase

b.Single strand binding proteins

c.Topoisomerase

d.DNA polymerase III

e.DNA polymerase I

f.Primase

g.Ligase

4)What extra steps are taken to be sure the lagging strand does not shorten each time DNA is replicated?

a.What is the role of telomerase in these âstepsâ?

5)In DNA repair, which enzyme has âproof-readingâ capabilities?

1)Which type of cells within our bodies use mitosis to reproduce?

2)Does mitosis create a cell that is different or the same as the parent cell undergoing division?

3)Describe the DNA found in sister chromatids.

4)Describe the DNA found in homologous chromosomes.

5)What occurs during the following stages of the cell cycle?

a.G₁ Phase

b.S Phase

c.G₂ Phase

d.M Phase

What are the phases of mitosis? What events occur in each phase?

1)Does mitosis separate homologous chromosomes, sister chromatids, or both?

2)What is cytokinesis?

a.Which fibers of the cytoskeleton contract to pinch parent and daughter cells apart from one another in animals?

b.How do microtubules move chromosomes to the two poles of a dividing cell?

3)How are centrioles involved in this process?

a.How do plants form two complete cells during cytokinesis?

4)How do bacterial cells reproduce?

5)The cell cycle is controlled at âcheckpointsâ. What occurs at each of the following checkpoints?

a.G₁ Checkpoint

b.G₂ Checkpoint

c.M Checkpoint

6)How do cyclin and cdk act to control the G2 checkpoint?

7)If presented with the schematic of the M checkpoint, you should be able to explain perturbations to the system as well as if the cell will divide or stall at metaphase.

8)Cancerous cells form when the cell cycle checkpoints do not function correctly and continues to divide even though it is damaged in some way. How does the protein p53 function to keep these cells from dividing?

9)Normal cells follow certain rules of growth that cancerous cells do not follow. What are these rules (provide a short definition for each as well)?

10)What is the difference between a benign tumor and a malignant tumor?

indigohedgehog242

1)What are the inputs of photosynthesis?

2)What are the outputs of photosynthesis?

3)Light-Dependent Reactions

Where does the first stage of photosynthesis, the light-dependent reactions, occur within a chloroplast?

Where does the energy come from that drives these reactions?

What molecule donates an electron to the reaction center of photosystem II?

How is the energy of an electron changed as it moves along the ETC?

How does the electrical charge of the thylakoid membrane affect protons (H+) within the chloroplast?

Where does the electron that is donated to photosystem I come from?

What are the final products of this series of reactions?

What happens to each of the final products from this series of reactions?

4)Light-Independent Reactions (Calvin Cycle)

a.Where does the second stage of photosynthesis, the Calvin Cycle, occur within a chloroplast?

b.What are the direct inputs for these reactions?

c.What are the three steps to the Calvin cycle?

d.What are the final products of this reaction?

e.What role does rubisco play in the Calvin Cycle?

1)What experiment was performed that allowed for scientists to finally agree that DNA is the genetic material within living organisms?

a.Understand the materials and scientific techniques used to come to this result.

2)What group of experiments led to the discovery of the mechanism used in DNA replication?

a.Understand the materials and scientific techniques used to come to this result.

i.What is a cesium chloride gradient?

ii.Where do 14N and 15N respectively fall along this gradient?

b.The results revealed that DNA replicates using which of the three proposed models?

i. Describe the mechanism of each model.

3)What is the role of the following proteins in DNA replication?

a.Helicase

b.Single strand binding proteins

c.Topoisomerase

d.DNA polymerase III

e.DNA polymerase I

f.Primase

g.Ligase

4)What extra steps are taken to be sure the lagging strand does not shorten each time DNA is replicated?

a.What is the role of telomerase in these âstepsâ?

5)In DNA repair, which enzyme has âproof-readingâ capabilities?

1)Which type of cells within our bodies use mitosis to reproduce?

2)Does mitosis create a cell that is different or the same as the parent cell undergoing division?

3)Describe the DNA found in sister chromatids.

4)Describe the DNA found in homologous chromosomes.

5)What occurs during the following stages of the cell cycle?

a.G₁ Phase

b.S Phase

c.G₂ Phase

d.M Phase

What are the phases of mitosis? What events occur in each phase?

1)Does mitosis separate homologous chromosomes, sister chromatids, or both?

2)What is cytokinesis?

a.Which fibers of the cytoskeleton contract to pinch parent and daughter cells apart from one another in animals?

b.How do microtubules move chromosomes to the two poles of a dividing cell?

3)How are centrioles involved in this process?

a.How do plants form two complete cells during cytokinesis?

4)How do bacterial cells reproduce?

5)The cell cycle is controlled at âcheckpointsâ. What occurs at each of the following checkpoints?

a.G₁ Checkpoint

b.G₂ Checkpoint

c.M Checkpoint

6)How do cyclin and cdk act to control the G2 checkpoint?

7)If presented with the schematic of the M checkpoint, you should be able to explain perturbations to the system as well as if the cell will divide or stall at metaphase.

9)Normal cells follow certain rules of growth that cancerous cells do not follow. What are these rules (provide a short definition for each as well)?

10)What is the difference between a benign tumor and a malignant tumor?

amberkoala41

HELP ASAP; Please help me understand my lab results, so be detailed to clearly understand

Then , Please include answers to the question below to include in your response

1. what role does ampicilin (amp) and arabinose (ara) play in the experiment.

2) Because GFP was isolated from jelly fisht, what modification to the GFP DNA had to be made to result in expression in E. coli?

BACKGROUND INFORMATION

Transformation of GFP and the pGLO Plasmid GFP is a commonly used reporter protein in research labs, as the fluorescence creates a marker protein that can be used in many types of cell biology and biochemical studies. In basic research, GFP is often fused to a specific target protein of interest, creating a chimeric reporter protein. GFP has been used as a reporter protein to study blood vessel and tumor progression in mice, brain activity in mice, and malaria eradication in mosquitoes for instance (see website mentioned in notes). In the first week you will use a procedure to genetically transform bacteria with the gene that codes for Green Fluorescent Protein. Following the transformation procedure, the bacteria express their newly acquired jellyfish gene and produce the fluorescent protein, which causes them to glow a brilliant green color under ultraviolet light. During the second week you will purify the protein away from all of the other proteins produced by the bacteria using chromatography, and during the third week you will evaluate the success of the purification procedure and estimate the size (molecular weight) of GFP by using sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE). Fundamental to the process of genetic transformation of bacteria are plasmids. In addition to one large chromosome, bacteria naturally contain one or more small circular pieces of DNA called plasmids. Plasmid DNA usually contains genes for one or more traits that may be beneficial to bacterial survival. In nature, bacteria can transfer plasmids back and forth allowing them to share these beneficial genes. This natural mechanism allows bacteria to adapt to new environments. The occurrence of bacterial resistance to antibiotics is due in large part to the transmission of plasmids which contain antibiotic resistance genes. The pGLO plasmid contains the gene for GFP, and also contains the gene for the enzyme Î²-lactamase (bla), which provides resistance to the antibiotic ampicillin. The Î²-lactamase protein is produced and secreted by bacteria that contain the plasmid. Î²-Lactamase inactivates the ampicillin present in the LB nutrient agar, allowing bacterial growth. Only transformed bacteria that contain the plasmid and express Î²-lactamase can survive on plates that contain ampicillin. Only a very small percentage of the cells take up the plasmid DNA and are transformed. Untransformed cells cannot grow on the ampicillin selection plates. pGLO also incorporates a special gene regulation system, which can be used to control expression of the fluorescent protein in transformed cells. The gene for GFP can be switched on in transformed cells by adding the sugar arabinose to the cellsâ nutrient medium. Selection for cells that have been transformed with pGLO DNA is accomplished by growth on antibiotic plates. Transformed cells will appear white (wild-type phenotype) on plates not containing arabinose, and fluorescent green when arabinose is included in the nutrient agar medium.

The transformation procedure involves three main steps. These steps are intended to introduce the plasmid DNA into the E. coli cells and provide an environment for the cells to express their newly acquired genes. 1) Cells are first treated with a solution of CaCl2, which is thought to neutralize the repulsive negative charges of the phosphate backbone of DNA and the phospholipids of the cell membrane, allowing the DNA to adhere to the cells. This is referred to making the cells âcompetentâ (i.e. the cells are capable, or competent, to take-in exogenous DNA). 2) Cells are then subjected to a heat shock, which is thought to increase the permeability of the cell membrane, allowing the cells to take-in the plasmid DNA. 3) Cells are allowed to grow in a ârecoveryâ phase in the absence of ampicillin. During this time the cells begin to produce the Î²-lactamase protein, which will allow them to survive when they are subsequently placed on agar plates containing ampicillin.

For the first week section of lab

To eac

h of two microfuge tubes, add 250ul of transformation buffer. Keep these tubes on ice.

Using a sterile loop, pick one colony of bacteria from the LB plate. Transfer this colony to one of the microfuge tubes.

Gently vortex the tube until the colony is dispersed. Repeat for the second tube. Add 0.08ug of DNA to one of these tubes (+DNA tube).

Incubate both tubes on ice for 10 minutes. Incubate both tubes at 42Â°C for 50 seconds.

Incubate both tubes on ice for 2 minutes. Add 250ul of LB broth to each tube and incubate at room temperature for 10 minutes. Flick each tube gently. Add 100ul of t

his transformation mix to the appropriate plates.

Results For plates:

+DNA: Has the E. Coli pGlo plasmid and - DNA plates do no have EColi pGLo plasmid .

Amp means ampiclin and Ara means arabinose on agar plates

+ DNA Plate LB/amp/ara--> white 9 colonies, Glow: yes

+DNA LB/amp--> 7 colonies, GLow: no

--DNA LB/AMP- 1 colony so no colony growth and no glovw

-DNA: LB--> many colonies, Glow: no

Please explain significance of result or thand the reason for the results in scientific terms based on the background information provided and your own knowledge to be detailed. Would these results be expected as wll.

BIO 1140 Chapter Notes - Chapter 12.3: Prophase, Receptor Tyrosine Kinase, Growth Medium

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