Consideration 1 can i genetically transform an organism

On which of the plates would you expect to find bacteria most like the original non-transformed E. Explain your predictions. If there are any genetically transformed bacterial cells, on which plate s would they most likely be located?

Which plates should be compared to determine if any genetic transformation has occurred? What is meant by a control plate? What purpose does a control serve? Data Collection Observe the results you obtained from the transformation lab under normal room lighting. Then turn out the lights and hold the ultraviolet light over the plates. Carefully observe and draw what you see on each of the four plates. Put your drawings in the data table in the column on the right.

Write down the following observations for each plate. How much bacterial growth do you see on each plate, relatively speaking? What color are the bacteria? How many bacterial colonies are on each plate count the spots you see. Analysis of Results The goal of data analysis for this investigation is to determine if genetic transformation has occurred. Which of the traits that you originally observed for E.

In the space below list these untransformed traits and how you arrived at this analysis for each trait listed. Original trait Analysis of observations 2. Of the E. List those traits below and describe the changes that you observed. New trait Observed change 3. If the genetically transformed cells have acquired the ability to live in the presence of the antibiotic ampicillin, then what might be inferred about the other genes on the plasmid that you used in your transformation procedure?

From the results that you obtained, how could you prove that the changes that occurred were due to the procedure that you performed? If a fluorescent green color is observed in the E. Explain: 1. Recall what you observed when you shined the UV light onto a sample of original pglo plasmid DNA and describe your observations.

Which of the two possible sources of the fluorescence can now be eliminated? What does this observation indicate about the source of the fluorescence? Describe the evidence that indicates whether your attempt at performing a genetic transformation was successful or not successful. Do you observe some E. From your results, can you tell if these bacteria are ampicillin resistant by looking at them on the LB plate? Explain your answer. How would you change the bacteria s environment the plate they are growing on to best tell if they are ampicillin resistant?

Very often an organism s traits are caused by a combination of its genes and its environment. Think about the green color you saw in the genetically transformed bacteria: a. What two factors must be present in the bacteria s environment for you to see the green color? Hint: one factor is in the plate and the other factor is in how you look at the bacteria.

What do you think each of the two environmental factors you listed above are doing to cause the genetically transformed bacteria to turn green? What advantage would there be for an organism to be able to turn on or off particular genes in response to certain conditions?

This quantitative measurement is referred to as the transformation efficiency. In many experiments, it is important to genetically transform as many cells as possible. For example, in some types of gene therapy, cells are collected from the patient, transformed in the laboratory, and then put back into the patient.

The more cells that are transformed to produce the needed protein, the more likely that the therapy will work. The transformation efficiency is calculated to help scientists determine how well the transformation is working.

The Task You are about to calculate the transformation efficiency, which gives you an indication of how effective you were in getting DNA molecules into bacterial cells. Transformation efficiency is a number. It represents the total number of bacterial cells that express the green protein, divided by the amount of DNA used in the experiment.

It tells us the total number of bacterial cells transformed by one microgram of DNA. Each colony on the plate can be assumed to be derived from a single cell. As individual cells reproduce, more and more cells are formed and develop into what is termed a colony. The most direct way to determine the total number of green fluorescent cells is to count the colonies on the plate.

This means that each microliter of solution contained 0. Calculate the total amount of DNA used in this experiment. Look in the laboratory procedure and locate all the steps where you added liquid to the reaction tube. Add the volumes. Decide which of the numbers you calculated belong in the table below. Fill in the following table. Scientists often use a mathematical shorthand referred to as scientific notation.

Report your calculated transformation efficiency in scientific notation. Use a sentence or two to explain what your calculation of transformation efficiency means. Biotechnologists are in general agreement that the transformation protocol that you have just completed generally has a transformation efficiency of between 8.

How does your transformation efficiency compare with the above? In the table below, report the transformation efficiency of several of the teams in the class. Team Efficiency How does your transformation efficiency compare with theirs? DNA plasmid concentration: 0. You can assume that the concentration of DNA and fraction of cells spread on the LB agar are the same as that of the pglo laboratory. Remember that a gene is a piece. Name: Transforming E. Coli with pglo Plasmids AP Biology Transformation Background: Transformation is a process of transferring genetic information from one organism to another.

In bacteria, a small circular. Student Manual pglo Transformation Lesson 1 Introduction to Transformation In this lab you will perform a procedure known as genetic transformation. Remember that a gene is a piece of DNA which provides. Pate Justin Rousseau Dohee Won Introduction The purpose of this experiment is to perform a genetic transformation and. Fluorescent Protein Transformation Student Background Genetic transformation occurs when a cell takes up i.

Genetic transformation literally. Outlines: 1-Insertion of foreign gene to the plasmid. Calculating the efficiency of DNA transfer during genetic engineering Specification references 3.

The plasmid. The key player in transduction. Amgen Protocol: Introduction and a few comments: The following is a shortened version of the Amgen Lab. Immerse a new sterile loop into the pGLO plasmid DNA stock tube and withdraw a loopful so that there is a film of the solution across the ring. Close the tube and return in to the rack on ice. Incubate the tubes on ice for 10 minutes. Make sure to push the tubes all the way down in the rack so that the tubes make contact with the ice.

Make sure that the tubes are all the way in the rack so that they touch the warm water. After 50 seconds, place both tubes back on ice. Incubate the tubes on ice for 2 minutes. Remove the rack containing the tubes from the ice and place them on the bench top. Repeat with a new sterile pipet for the other tube. Incubate the tubes for 10 minutes at room temperature. Tap the closed tubes with your finger to mix. Use a new sterile loop for each plate.

Spread the suspension evenly around the surface of the LB nutrient agar by quickly skating the flat surface of a new sterile loop back and forth across the plate surface. To ensure that the individuals demonstrating the lab are not physically affected by the contents of their work, having an experimental organism that is not toxic to the body is crucial. The organisms best suited for genetic transformation are bacterium.

Bacteria are well suited for genetic modification as they consist of a single cell and will become transgenic organisms after being mixed with DNA. In addition only one cell needs to be changed in order to incorporate the new transgenic information and allow for its transmission to the next generation. Athira You could have one plate as the positive control with some E.

You can then incubate them and see how E. What would you expect your experimental results to indicate about the effect of ampicillin on the E.

Athira One would expect to see the E. This is because an organism with one cell only needs to take the new gene into one cell which is more efficient and faster than inserting the gene into every organism.

To get this information, which would be a better candidate for your investigation, an organism in which new generation develops and reproduces quickly, or one which does this more slowly?

The better candidate would be the organism in which the new generation develops and reproduces quickly. This is because the quicker the organism reproduces, the scientist can see if the trait was actually passed on to the next generation or if modifications need to be made. What traits or characteristics should the organism have or not have to be sure it will not harm you or the environment? The organism should not be antibiotic resistant because if it were antibiotic resistant, and the scientist were infected, antibiotics could not be used to treat the disease.

Also the organism should not be UV resistant because UV is sometimes used to kill the organism.