Table of Contents
What is the pGLO lab?
pGLO Bacterial Transformation and GFP Kits pGLO Lab Kits utilize Bio-Rad’s pGLO plasmid, which encodes a green fluorescent protein (GFP), to enable instructors to give students a hands-on introduction to transformation, cloning, protein chromatography, and electrophoresis techniques.
What does the pGLO name refer to?
STUDY. plasmid. A small circular piece of DNA.
What does LB stand for pGLO lab?
LB: luria broth-nutrients. Amp: the antibiotic ampicillin. Ara: the sugar arabinose. +pGLO: contains transformed bacteria. -pGLO: contains non-transformed bacteria.
What does pGLO stand for in biology?
PGLO does not stand for something specific and is referred to as ‘pGLO plasmid’. This plasmid is used to transfer, transform, and express bacteria….
What is the purpose of the pGLO experiment?
They have been used to figure out how certain bacteria reacts to different types of antibiotics and to change how bacteria functions by adding things into the plasmids. In this lab the plasmid that we focused on was the pGLO plasmid.
What is pGLO quizlet?
What is pGLO? a type of genetically engineered plasmid that encodes the gene for GFP and a gene for resistance to the antibiotic ampicillin. Also incorporates a special gene regulation system that can be used to control expression of the fluorescent protein in transformed cells.
What does the pGLO name refer to quizlet?
What is pGLO? a type of genetically engineered plasmid that encodes the gene for GFP and a gene for resistance to the antibiotic ampicillin.
What is pGLO transformation?
The pGLO System With the pGLO transformation kit, students use a simple procedure to transform bacteria with a gene that codes for Green Fluorescent Protein (GFP). The real-life source of this gene is the bioluminescent jellyfish Aequorea victoria, and GFP causes the jellyfish to fluoresce and glow in the dark.
What does LB amp and ARA stand for?
LB/amp/ara (Luria Broth + ampicillin + arabinose): on which only transformed E coli grow. They do fluoresce as the arabinose in the medium causes the promoter to switch on the gene for GFP.
What is E. coli pGLO?
in the transformation of the E. coli is the pGLO plasmid, this plasmid contains the ampicillin resistance gene and the GFP gene. If the bacteria was able tosuccessfully be transformed with the plasmid, the bacteria will contain the two genes. The E. coli will be ampicillin resistant and will produce GFP.
How is pGLO plasmid made?
This recombinant plasmid, created by researchers at Bio-Rad, combines a gene for green fluorescent protein (GFP), cloned from a jellyfish, with control elements copied from a bacterial operon. The end result is a system that allows for bacterial expression of a eukaryotic gene.
What are the 3 genes of interest on the pGLO plasmid?
The pGLO plasmid contains both the promoter (pBAD) and araC gene, but araB, araA, and araD have been replaced by the single gene that codes for GFP, which serves as a reporter gene.
How does the pGLO bacterial transformation relate to Griffith’s mice experiment?
when a mouse was injected with nonvirulent and heat-killed virulent bacteria the mice did die. showing that something from the dead bacteria caused the nonvirulent bacteria to kill the host.
What is LB amp?
LB Amp is Lysogeny Broth (LB) containing the antibiotic ampicillin. Because LB is a rich medium for growing bacteria, adding ampicillin provides a means of selecting transformants that have taken up plasmid DNA containing the bla gene, which encodes resistance to ampicillin.
Which plates glow in the pGLO lab?
The Green Fluoresce Protein (GFP) is a gene code that is inserted into the pGLO plasmid to make it glow green under UV light with the help of the araC gene….pGLO Lab Analysis.
| DNA | Type of plate | Prediction |
|---|---|---|
| pGLO wild-type | Ampicillin + Arabinose | Green colonies |
| pGLO mutagenized | Kanamycin + Arabinose | Green + White colonies |
Why was the pGLO LB plate included in the experiment?
The purpose of spreading pGLO on the LB plate was to enable bacteria to grow, otherwise the ampicillin would kill them. If the pGLO is not spread, then the cells would be killed by the ampicillin.
What is in the pGLO plasmid?
The pGLO plasmid contains an origin or replication, a selectable marker, and the gene for Green Fluorescent Protein (GFP). The plasmid also contains a gene for the arabinose C protein, which is a protein that regulates expression from the arabinose BAD promoter (PBAD).
What does pGLO do to bacteria?
The pGLO System 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.
What is PGLO and how does it work?
A description of what you’ll be working with, including a brief overview of transformation. pGLO: How the plasmid works. In this lab, you’ll transform E. coli cells with the plasmid pGLO, which contains a gene for green fluorescent protein (GFP). This will allow you to grow bacterial colonies that are bright, fluorescent green.
What is a pGLO plasmid?
Plasmids have been studied in genetic research since they were discovered in 1952 by Joshua Lederberg. They have been used to figure out how certain bacteria reacts to different types of antibiotics and to change how bacteria functions by adding things into the plasmids. In this lab the plasmid that we focused on was the pGLO plasmid.
Can pGLO survive ampicillin?
pGLO Lab Analysis. Only transformed bacteria that contain the plasmid and express beta-lactamase can survive on plates that contain ampicillin.” (Leatherman 2011). The Green Fluoresce Protein (GFP) is a gene code that is inserted into the pGLO plasmid to make it glow green under UV light with the help of the araC gene.
What is the function of the PBAD promoter in pGLO?
The GFP operon in pGLO uses the pBAD promoter from E. coli. In wild bacteria, the pBAD promoter controls the expression of the AraBAD operon, which encodes genes coding for proteins (called AraB, AraA, and AraD) involved in catabolism of the sugar arabinose.