Malcolm Rivera
The pGLO plasmid is a powerful tool in biotechnology, enabling the creation of genetically modified organisms. It is a circular piece of DNA engineered using recombinant DNA technology. The pGLO plasmid contains several genes, including the green fluorescent protein (GFP) gene, which is expressed in the presence of arabinose, causing the transgenic organism to fluoresce under UV light. The GFP gene is regulated by the araC gene, which encodes a protein that binds to the pBAD promoter and activates GFP expression. Additionally, the pGLO plasmid contains the bla gene, which confers resistance to antibiotics like ampicillin. This plasmid is commonly used in classrooms and laboratories to teach gene regulation and expression, bacterial transformation, and other fundamental concepts in biotechnology and genetic engineering.
| Characteristics | Values |
|---|---|
| Purpose | To demonstrate genetic exchange in bacteria and eukaryotes |
| Genes | GFP, Bla, araC |
| Gene Expression | GFP is expressed in the presence of arabinose |
| Gene Function | GFP: Green fluorescent protein; Bla: Encodes beta-lactamase; araC: Encodes a regulatory protein |
| Gene Source | GFP is isolated from the jellyfish Aequorea victoria |
| Gene Regulation | araC regulates the expression of GFP |
| Gene Selection | araC acts as a selectable marker |
| Gene Transformation | CaCl2/heat shock method is used to transform E. coli with the pGLO plasmid |
| Gene Replication | The pGLO plasmid contains an origin of replication (ori) |
| Gene Insertion | The pGLO plasmid contains a multiple cloning site (MCS) |
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What You'll Learn
The pGLO plasmid is used to create genetically modified organisms
The pGLO plasmid is an engineered plasmid used in biotechnology as a vector for creating genetically modified organisms. The plasmid contains several reporter genes, most notably the green fluorescent protein (GFP) and the ampicillin resistance gene. GFP was first observed in 1962 by Osamu Shimomura and his team while studying the jellyfish *Aequorea victoria*, which has a ring of blue light under its umbrella.
The pGLO plasmid is designed to enable replication of the plasmid in bacteria, selection of transformants using ampicillin resistance, and regulation of GFP expression. The GFP gene is expressed in the presence of arabinose, which makes the transgenic organism express its fluorescence under UV light. GFP can be induced in bacteria containing the pGLO plasmid by growing them on +arabinose plates.
Genetic transformation is the process by which an organism acquires and expresses a new gene. Genetic engineering is the directed transfer of a gene or piece of DNA into a cell. The pGLO plasmid is used to introduce foreign DNA into bacterial cells, enabling scientists to manipulate and study genes, gene function, and gene expression. The pGLO Bacterial Transformation Kits use the CaCl2/heat shock method to enable bacterial transformation with the pGLO plasmid. This method can be completed with no additional laboratory equipment beyond insulated cups, warm water, and ice.
The pGLO plasmid is a fantastic tool for teaching gene regulation in a hands-on and engaging way. The pGLO Bacterial Transformation and GFP Kits offer engaging and unforgettable student lab activities in which students engineer bacteria to express the Green Fluorescent Protein (GFP) and glow green under UV light.
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The plasmid contains the gene encoding GFP
The pGLO plasmid is an engineered plasmid used in biotechnology as a vector for creating genetically modified organisms. The plasmid contains several reporter genes, most notably the gene encoding green fluorescent protein (GFP) and the ampicillin resistance gene. GFP was originally isolated from the jellyfish *Aequorea victoria*.
The pGLO plasmid is designed to enable replication of the plasmid in bacteria, selection of transformants using ampicillin resistance, and regulation of GFP expression. The GFP gene is expressed in the presence of arabinose, which makes the transgenic organism express its fluorescence under UV light. GFP can be induced in bacteria containing the pGLO plasmid by growing them on +arabinose plates.
The pGLO plasmid contains the gene encoding GFP, which stands for green fluorescent protein. This gene is under the control of a promoter from the arabinose operon, and so transcription occurs in the presence of this sugar. The GFP gene was first observed by Osamu Shimomura and his team in 1962 while studying the jellyfish *Aequorea victoria* that have a ring of blue light under their umbrella.
The pGLO plasmid is commonly used in classrooms to teach students about gene expression and regulation, bacterial transformation, and the biomanufacturing process. The pGLO Bacterial Transformation Kits enable students to perform bacterial transformation and express GFP in a non-virulent laboratory strain of Escherichia coli (E. coli). This is achieved by introducing foreign DNA into bacterial cells, which enables the manipulation and study of genes, gene function, and gene expression.
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The GFP gene is expressed in the presence of arabinose
The pGLO plasmid is an engineered plasmid used in biotechnology as a vector for creating genetically modified organisms. The plasmid contains several reporter genes, most notably the green fluorescent protein (GFP) and the ampicillin resistance gene. GFP was first observed by Osamu Shimomura and his team in 1962 while studying the jellyfish Aequorea victoria, which has a ring of blue light under its umbrella.
The pGLO plasmid is designed to enable replication of the plasmid in bacteria, selection of transformants using ampicillin resistance, and regulation of GFP expression. Bacteria transformed with the pGLO plasmid are selected by ampicillin resistance and then induced to express GFP to glow fluorescent green under UV light.
The GFP gene shares a bidirectional promoter with a gene for metabolizing arabinose. The concentration of arabinose in the growth medium can regulate gene expression from plasmids containing the araBAD promoter. Lower concentrations of arabinose led to lower steady-state levels of GFP in the cultures. High-level expression of GFP seems to be toxic to E. coli; cells plated on Luria–Bertani medium with 0.2% arabinose in the absence of ampicillin form colonies with a variable number of dark and fluorescent sectors. In the absence of arabinose, or in the presence of glucose, the cells are uniformly dark under UV illumination.
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The pGLO plasmid also contains a gene for ampicillin resistance
The pGLO plasmid is an engineered plasmid used in biotechnology as a vector for creating genetically modified organisms. The plasmid contains several reporter genes, most notably the gene for green fluorescent protein (GFP) and the gene for ampicillin resistance.
Ampicillin is an antibiotic for gram-negative cells such as E. coli, which is part of the penicillin family of antibiotics. It inhibits the production of the outer cell membrane/proteoglycan structure, which is lethal to the bacteria. Ampicillin, as a molecule, contains a beta-lactam ring structure that can be cleaved by the enzyme beta-lactamase. The gene for beta-lactamase is called bla or ampR, and it is present in the pGLO plasmid. This gene confers resistance to ampicillin to the host cells.
The pGLO plasmid is used in classrooms to teach students about gene expression and regulation, bacterial transformation, protein separation, and the biomanufacturing process. The Bio-Rad pGLO Bacterial Transformation Kits enable students to perform bacterial transformation and express GFP in a non-virulent laboratory strain of Escherichia coli (E. coli). The pGLO plasmid is designed to enable the replication of the plasmid in bacteria, the selection of transformants using ampicillin resistance, and the regulation of GFP expression.
The selection of transformants using ampicillin resistance is achieved by growing the bacteria on a medium containing ampicillin. Only the bacteria that have successfully taken up the pGLO plasmid, and therefore have the gene for ampicillin resistance, will survive and grow in the presence of ampicillin. This aspect of the system can allow for the exploration of antibiotic structure and enzyme specificity.
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The pGLO plasmid is a circular piece of DNA
The pGLO plasmid is made up of three genes that are joined together using recombinant DNA technology. The first gene, *bla*, codes for the enzyme beta-lactamase, which breaks down antibiotics like penicillin. This gene confers resistance to the beta-lactam family of antibiotics, including ampicillin. Ampicillin is an antibiotic for gram-negative cells such as E. coli, and it inhibits the production of the outer cell membrane, which is lethal to the bacteria.
The second gene, *araC*, is a promoter region that regulates the expression of the GFP gene. Specifically, the GFP gene will only be expressed in the presence of arabinose. The addition of arabinose sugar to the growth media causes RNA polymerase to start transcribing the GFP gene. The GFP gene is expressed in the presence of arabinose, which makes the transgenic organism express its fluorescence under UV light.
The third gene is the GFP gene, which stands for green fluorescent protein. When this gene is turned on, it causes the bacteria to glow green under UV light. This allows scientists to visually confirm that the bacteria have taken up the plasmid.
The pGLO plasmid also contains an origin of replication (*ori*) that allows the plasmid to replicate inside bacteria, and a multiple cloning site (MCS) where additional DNA can be inserted. The pGLO plasmid is commonly used in classrooms and laboratories to teach gene regulation and genetic transformation in a hands-on and engaging way.
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