BIO SCI 93 Study Guide - Prophase, Meiosis, Dapi

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BIO SCI 93 Full Course Notes

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Bio 93 problem set 3 unit iv: dividing the cell. Due wed. nov. 6 at 5 pm, upload as a pdf: you are culturing a human leukemia cell line as a part of your undergraduate research project. You count the cells in your flask at 8 am each morning when you get to lab and record the data in your notebook. Monday, you had 10 million cells in your flask and on tuesday you have 20 million cells. Your supervisor wants to know how long it takes this cell line to go through mitosis. It takes one day for the cell to go through mitosis. Using a fluorescence microscope, you can see only the microtubules in each cell glowing on a black background. Draw what the microtubules would look like in one of these cells in the following cell cycle stages (a cell outline is provided but would not be visible under the microscope):

Related Questions

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.

HELP ASAP;

Please help me understand my lab results, so be detailed to clearly understand . INclude answers to these questions what role does ampicilin (amp) and arabinose (ara) play in the experiment. 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 (Fig. 5, not a naturally occurring 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.

for the first week section of lab

To each 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 this transformation mix to the appropriate plates.

Results:

+DNA: LB/amp--> 1 colony, Glow: uncertain

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

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

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

Which of the two major categories of the chemical reactions in photosynthesis is oxygen the INDIRECT product?

Glycolysis

Oxidative phosphorylation

Both light and dark

Light independent phase (LIP)

Light dependent phase (LDP)

You have a stomach flu characterized by vomiting. Which of the following systems is most responsible for the food in your stomach going the wrong way, e.g. vomiting? (Disregard this question and answer choice B)

Cardiac Muscle in the large intestine as discussed in class.

Smooth muscle in the gastrointestinal tract

Smooth muscle in the blood vessels

Skeletal muscle in the intestine

Proteins are made from which of the following smaller sub units?

Nucleotides

Amino acids that cannot enter the blood stream and go to cells

Monosaccharides

Fatty acids

Amino acids which can form new protein

Carbohydrates are digested in the gastrointestinal system into which of the following substances that is the smallest structure?

Polysaccharides that are produced by glucagon

Monosaccharides that are produced by insulin

Monosaccharides

Polysaccharide

Which of the following biomolecules has the longest digestion time? In other words, how long does it take for the biomolecule to be catabolized?

DNA

Lipid

Carbohydrate

Protein

You are sunburned after a long day at the beach. Which one is not an example of UV damage to your skin cells?

A mutation can occur in your ovaries

A mutation can occur in skin cell DNA that can lead to errors in transcription.

Errors can occur during DNA replication.

UV light can cause mutations in DNA of skins cells.

Thymine bases in your skin DNA bond with each other instead of across the double helix

This region, the ______________ (a) tells RNA polymerase where to bind to DNA while this region, the ______________ (b) tells RNA where to start transcription.

(a) promoter, (b) open reading frame

(a) start codon, (b) methionine

(a) promoter, (b) start site

(a) start site, (b) promoter

The genetic code is used to translate ______________ into ______________.

lipids, nucleotides

nucleotides,carbohydrates

nucleotides, lipids

nucleotides, amino acids

nucleosides, amino acids

Transcription of a gene has just completed. What are the next steps (in the correct order) to complete expression of that gene?

RNA polymerase makes a copy of DNA into mRNA.

mRNA is translated in the nucleus and ribosomal RNA moves into the cytoplasm.

Gene expression is completed after transcription.

mRNA moves into the cytoplasm and with tRNA they interact to form protein

Translation machinery moves into the nucleus and mRNA is translated into protein.

How is it that a mutation in DNA stored inside the nucleus of a cell can cause a disease affecting multiple organs?

This remains an unanswered question in science that scientists are still trying to figure out.

The mutated DNA is expressed in multiple cells through the process of transcription and translation, and the altered protein causes the disease wherever the DNA was expressed.

The mutated DNA is expressed in one cell and the altered protein travels to multiple organs to cause disease.

The mutated DNA is expressed and the mRNA takes the mutated information to other cells where the altered protein is made to cause disease in that organ.

The advantage of sickle cell anemia is which of the following?

It decreases the amount of oxygen in the blood cells so that mosquitoes carrying malaria will live

It decreases the amount of oxygen in the blood cells so that mosquitoe's blood form will die.

It allows the blood vessels to decrease the amount of blood going to the toes and feet.

There is none.

Which of the following is a purpose of cell division?

Cell reproduction

Growth

Repair

All of these

BIO SCI 93 Study Guide - Prophase, Meiosis, Dapi

BIO SCI 93 Full Course Notes

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