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Questions and Answers
Which process involves bacteria taking in plasmids from their environment?
Which process involves bacteria taking in plasmids from their environment?
- Binary Fission
- Transformation (correct)
- Transduction
- Conjugation
What characterizes competent cells in bacterial transformation?
What characterizes competent cells in bacterial transformation?
- Cells that reproduce sexually
- Cells that can take up plasmids (correct)
- Cells capable of photosynthesis
- Cells that are resistant to all antibiotics
What is the role of arabinose in the context of the pGLO gene?
What is the role of arabinose in the context of the pGLO gene?
- It increases bacterial mutation rates
- It induces the expression of the pGLO gene (correct)
- It acts as a selective marker
- It is necessary for conjugation
Which method would be used to make bacteria artificially competent?
Which method would be used to make bacteria artificially competent?
Which of the following describes transduction?
Which of the following describes transduction?
What type of gene does the pGLO plasmid confer resistance to?
What type of gene does the pGLO plasmid confer resistance to?
In which of the following plates would you expect to see growth of bacteria containing the pGLO plasmid?
In which of the following plates would you expect to see growth of bacteria containing the pGLO plasmid?
What is the primary method of reproduction in bacteria?
What is the primary method of reproduction in bacteria?
What is the outcome of conjugation in bacteria?
What is the outcome of conjugation in bacteria?
What occurs during the heat shock step of the transformation protocol?
What occurs during the heat shock step of the transformation protocol?
Which of the following statements is true about horizontal gene transfer?
Which of the following statements is true about horizontal gene transfer?
What type of plates are used for the +pGLO tube after transformation?
What type of plates are used for the +pGLO tube after transformation?
What is the purpose of adding LB to both tubes after the heat shock step?
What is the purpose of adding LB to both tubes after the heat shock step?
What does the arabinose gene do in relation to the pGLO plasmid?
What does the arabinose gene do in relation to the pGLO plasmid?
What type of colonies are picked for the transformation process?
What type of colonies are picked for the transformation process?
Which of the following statements accurately describes the -pGLO tube?
Which of the following statements accurately describes the -pGLO tube?
Why is the heat shock procedure performed on the transformation solution?
Why is the heat shock procedure performed on the transformation solution?
What role does the gene for ampicillin resistance play in the experiment?
What role does the gene for ampicillin resistance play in the experiment?
What is the significance of incubating the tubes on ice prior to heat shock?
What is the significance of incubating the tubes on ice prior to heat shock?
What is the function of arabinose in the experiment described?
What is the function of arabinose in the experiment described?
What would be the predicted growth result of bacteria B on nutrient agar with vancomycin and streptomycin?
What would be the predicted growth result of bacteria B on nutrient agar with vancomycin and streptomycin?
Which media combination would NOT support the growth of bacteria B after conjugation with bacteria A?
Which media combination would NOT support the growth of bacteria B after conjugation with bacteria A?
What would be the expected result regarding pGFP expression in bacteria B after successful conjugation?
What would be the expected result regarding pGFP expression in bacteria B after successful conjugation?
What is the role of ampicillin in the media?
What is the role of ampicillin in the media?
What type of bacteria is Species B before conjugation?
What type of bacteria is Species B before conjugation?
If arabinose is present in the nutrient agar, what can be concluded about bacteria A?
If arabinose is present in the nutrient agar, what can be concluded about bacteria A?
What does the term 'conjugation' refer to in the context of this experiment?
What does the term 'conjugation' refer to in the context of this experiment?
What role does the chromosomal gene for mercury resistance play in the experiment?
What role does the chromosomal gene for mercury resistance play in the experiment?
What would be the consequence of not adding arabinose to the media with bacteria A?
What would be the consequence of not adding arabinose to the media with bacteria A?
Study Notes
Transformation
- Bacteria can acquire new genetic material through horizontal gene transfer
- Horizontal gene transfer: the transfer of genetic information between adult organisms
- Transformation: bacteria taking in plasmids from the environment
- Transduction: transfer of DNA via bacteriophages
- Conjugation: bacteria exchanging plasmids
- Competent cells: bacteria that can be transformed by plasmids
- Artificially competent cells: bacteria treated with heat shock to make them more permeable to plasmids
- Heat shock process:
- Salt/heat to soften the cell membrane and allow plasmids to enter
- Ice to stiffen the membrane and trap the plasmids inside
pGLO Plasmid
- pGLO plasmid contains two key genes:
- ampR gene: provides resistance to ampicillin
- pGLO gene: encodes the green fluorescent protein (GFP)
- The pGLO gene is controlled by the pBAD promotor
- pBAD promotor: requires arabinose to activate the pGLO gene
- Arabinose: sugar molecule that acts as an inducer for the pGLO gene
Transformation Experiment
- Aim: To transfer the pGLO plasmid containing the GFP gene into E. coli bacteria
- Procedure:
- Two tubes labeled "-pGLO" and "+pGLO"
- "-pGLO" tube contained E. coli bacteria only
- "+pGLO" tube contained E. coli bacteria and the pGLO plasmid
- Both tubes were subjected to heat shock
- Each tube was plated on four different agar plates:
- LB: nutrient-rich medium
- LB + Amp: LB with ampicillin
- LB + Amp + Ara: LB with ampicillin and arabinose
- Expected results:
- -pGLO plates:
- LB: bacterial growth
- LB + Amp: no growth (ampicillin resistant genes absent)
- +pGLO plates:
- LB + Amp: bacterial growth (ampicillin resistance due to plasmid)
- LB + Amp + Ara: bacterial growth and green fluorescence under UV light (GFP expression induced by arabinose)
- -pGLO plates:
Conjugation Experiment
- Aim: To transfer the pGLO plasmid from a donor bacterium (F+) to a recipient bacterium (F-)
- Donor bacterium (F+):
- Contains the pGLO plasmid
- pGLO plasmid: ampicillin resistance, arabinose-inducible GFP, and GFP gene
- Contains a chromosomal gene for mercury resistance
- Recipient bacterium (F-):
- Does not contain the pGLO plasmid
- Has chromosomal genes for streptomycin and vancomycin resistance
- Expected results for recipient bacterium after conjugation:
- Nutrient agar, +vanc, +strep: growth (recipient bacterium retains its original resistance)
- Nutrient agar, +vanc, +strep, +amp: growth (recipient bacterium acquires ampicillin resistance from the plasmid)
- Nutrient agar, +vanc, +strep, +amp, +arab: growth and fluorescence (recipient bacterium acquires the pGLO plasmid and expresses GFP in the presence of arabinose)
- Nutrient agar, +vanc, +strep, +amp, +merc: no growth (recipient bacterium does not acquire mercury resistance as it is encoded on the donor's chromosome)
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Description
This quiz explores essential concepts of horizontal gene transfer in bacteria, with a focus on transformation, transduction, and conjugation. Additionally, it covers the properties of the pGLO plasmid and its components such as the ampR and pGLO genes. Test your understanding of these important microbiological processes.
