Microbiology, Lab 6 pGLO Transformation of E. coli Test with Answers
1. Genetic Transformtion literally means: "change caused by genes."
-involves the insertion of some new DNA into the cell. (such as Plasmids)
2. Gene a piece of DNA which provides the instructions for making (codes for) a
protein.
3. Protein after genetic gives an organism a particular trait.
transformation
4. Genetic Transformation -agriculture: frost, pest, spoilage resistance
in Biotechnology world -Bioremediation: where bacteria can be transformed with genes that
give them certain abilities.
-medicine: gene therapy.
5. GFP green fluorescent protein: makes those with this gene glow in the dark.
Originates from organism: Bioluminescent jellyfish= Aequorea victoria.
6. Plasmid A small ring of DNA that carries accessory genes separate from those of
the bacterial chromosome.
Plasmids pass genetic information b/t bacteria.
Plasmid DBA usually contain genes for more than one trait.
7. genetic engineering used to insert genes coding for new traits into a plasmid.
here: plasmid has been engineered to carry the GFP gene.
8. What two factors code 1. GFP protein
for a protein that gives 2. bla (or amp) gene
bacteria resistance to an
antibiotic?
9. Bio-Rad's pGLO plasmid
,Microbiology, Lab 6 pGLO Transformation of E. coli Test with Answers
it encodes for the gene for GFP and a gene for resistance to the antibiotic
ampicillin.
pGLO also incorporates a special gene regulation system that can be
switched on in transformed cells by adding sugar arabinose to the cells'
nutrient medium.
Bio-Rad's pGLO plasmid contains DNA sequences that enable its repli-
cation and expression of the fluorescent trait (phenotype) in bacteria
following transformation.
10. Ampicillin plates selection for cells that have been transformed with pGLO DNA is accom-
plished by growth on these plates.
11. Bio-Rad's pGLO plasmid a. GFP: jellyfish gene that encodes for fluorescent protein (GFP).
essential sequences:
b. ori: origin of pGLO plasmid DNA replication. (essential for
making more copies of the plasmid.
-(remember "origin of replication)
c. bla: gene that encodes β-lactamase, an enzyme that breaks
down the antibiotic ampicillin, transformants expressing the bla gene can
be selected by placing ampicillin in the growth medium.
-An ampicillin-resistant gene ampr (or bla) is used to select for cells that
have been transformed.
-Transformed colonies growing on LB/amp/ara, will glow green due to
GFP expression.
d. pBAD promoter: binds AraC-arabinose and promotes RNA
polymerase binding and transcription of the GFP gene.
-pBAD promoter is controlling the expression of the gene for GFP on the
, Microbiology, Lab 6 pGLO Transformation of E. coli Test with Answers
plasmid.
e. araC: a gene that encodes the regulatory protein that binds
to the pBAD promoter, only when arabinose binds to the AraC protein
is the production of GFP switch in.
-An inhibitor protein AraC encoded by the araC gene inhibits transcrip-
tion (gene expression) by binding to the pBAD promoter unless the
sugar arabinose is present.
-When arabinose is present it will bind to the inhibitor and cause it to
change shape and leave the promoter.
-This is an inducible system like the lac operon.
-Only transformed cells that are growing on LB agar with arabinose will
express GFP.
d. Multiple cloning site: a region containing restriction sites, sequences
that permit the insertion or detection of a gene of interest.
12. Cell Colors transformed cells= White (due to wild-type phenotype)
-this occurs on plates w/o arabinose.
the cells glow green under UV light when arabinose is included in the
nutrient agar medium.
13. Satellite colonies: There is an amp gene (can also be called bla) for ampicillin resistance.
This gene encodes beta-lactamase, an enzyme that degrades penicillin
and ampicillin. This gene product is secreted by cells that have taken up
the plasmid.
This is what we are using to select for cells that have been transformed.
1. Genetic Transformtion literally means: "change caused by genes."
-involves the insertion of some new DNA into the cell. (such as Plasmids)
2. Gene a piece of DNA which provides the instructions for making (codes for) a
protein.
3. Protein after genetic gives an organism a particular trait.
transformation
4. Genetic Transformation -agriculture: frost, pest, spoilage resistance
in Biotechnology world -Bioremediation: where bacteria can be transformed with genes that
give them certain abilities.
-medicine: gene therapy.
5. GFP green fluorescent protein: makes those with this gene glow in the dark.
Originates from organism: Bioluminescent jellyfish= Aequorea victoria.
6. Plasmid A small ring of DNA that carries accessory genes separate from those of
the bacterial chromosome.
Plasmids pass genetic information b/t bacteria.
Plasmid DBA usually contain genes for more than one trait.
7. genetic engineering used to insert genes coding for new traits into a plasmid.
here: plasmid has been engineered to carry the GFP gene.
8. What two factors code 1. GFP protein
for a protein that gives 2. bla (or amp) gene
bacteria resistance to an
antibiotic?
9. Bio-Rad's pGLO plasmid
,Microbiology, Lab 6 pGLO Transformation of E. coli Test with Answers
it encodes for the gene for GFP and a gene for resistance to the antibiotic
ampicillin.
pGLO also incorporates a special gene regulation system that can be
switched on in transformed cells by adding sugar arabinose to the cells'
nutrient medium.
Bio-Rad's pGLO plasmid contains DNA sequences that enable its repli-
cation and expression of the fluorescent trait (phenotype) in bacteria
following transformation.
10. Ampicillin plates selection for cells that have been transformed with pGLO DNA is accom-
plished by growth on these plates.
11. Bio-Rad's pGLO plasmid a. GFP: jellyfish gene that encodes for fluorescent protein (GFP).
essential sequences:
b. ori: origin of pGLO plasmid DNA replication. (essential for
making more copies of the plasmid.
-(remember "origin of replication)
c. bla: gene that encodes β-lactamase, an enzyme that breaks
down the antibiotic ampicillin, transformants expressing the bla gene can
be selected by placing ampicillin in the growth medium.
-An ampicillin-resistant gene ampr (or bla) is used to select for cells that
have been transformed.
-Transformed colonies growing on LB/amp/ara, will glow green due to
GFP expression.
d. pBAD promoter: binds AraC-arabinose and promotes RNA
polymerase binding and transcription of the GFP gene.
-pBAD promoter is controlling the expression of the gene for GFP on the
, Microbiology, Lab 6 pGLO Transformation of E. coli Test with Answers
plasmid.
e. araC: a gene that encodes the regulatory protein that binds
to the pBAD promoter, only when arabinose binds to the AraC protein
is the production of GFP switch in.
-An inhibitor protein AraC encoded by the araC gene inhibits transcrip-
tion (gene expression) by binding to the pBAD promoter unless the
sugar arabinose is present.
-When arabinose is present it will bind to the inhibitor and cause it to
change shape and leave the promoter.
-This is an inducible system like the lac operon.
-Only transformed cells that are growing on LB agar with arabinose will
express GFP.
d. Multiple cloning site: a region containing restriction sites, sequences
that permit the insertion or detection of a gene of interest.
12. Cell Colors transformed cells= White (due to wild-type phenotype)
-this occurs on plates w/o arabinose.
the cells glow green under UV light when arabinose is included in the
nutrient agar medium.
13. Satellite colonies: There is an amp gene (can also be called bla) for ampicillin resistance.
This gene encodes beta-lactamase, an enzyme that degrades penicillin
and ampicillin. This gene product is secreted by cells that have taken up
the plasmid.
This is what we are using to select for cells that have been transformed.
