Exam 2 (Ch. 7, 8, & 9) Flashcards
What are some of the MAJOR DIFFERENCES between Prokaryotic and Eukaryotic Transcription and Translation?
EUKARYOTES:
1. A cap is added to the 5’ tail end of mRNA, and a poly A tail is added to the 3’ end
2. mRNA contains introns, which are removed by splicing
3. The mRNA transcript is transported out of the nucleus so that it can be translated in the cytoplasm
4. mRNA is monocistronic; translation begins at the first AUG
PROKARYOTES:
1. mRNA is NOT processed
2. mRNA DOES NOT contain introns
3. Translation of mRNA begins as it is being transcribed
4. mRNA is often polycistronic; translation usually begins at the first AUG that FOLLOWS a ribosome binding site
Are bacteria haploid or diploid?
Haploid
T/F: Bacterial toxins can target eukaroytic enzymes
True
does glucose or lactose gets exhausted first in the Lac Operon graph?
glucose
What is an operon?
a group of structural genes plus sequences that control transcription
includes structural genes, a promoter, and an operator
There are two mechanisms of regulation by repressors.
What occurs in Repressible Control? (e.g. trp)
TRANSCRIPTION ON:
-The repressor alone CANNOT bind to the operator, so transcription continues
TRANSCRIPTION OFF (BLOCKED):
–Corepressor binds to the repressor and ALTERS its shape, enabling it to bind to the operator and blocking transcription
There are two mechanisms of regulation by repressors.
What occurs in Indicible Control?
TRANSCRIPTION OFF (BLOCKED):
-Repressor binds to the operator and blocks transcription
TRANSCRIPTION ON:
-Inducer binds to the repressor and ALTERS its shape, preventing it from binding to the operator
During Lac operon, what happens in transcription when glucose is present, but not lactose?
Transcription not activated AND blocked. Low cAMP-, CAP cannot bind. repressor is bound to operator, blocking polymerase.
What is Positive Inducible Control when it comes to activating transcription?
TRANSCRIPTION OFF (NOT ACTIVATED:
-Activator CANNOT bind to the activator-binding site, thus RNA polymerase cannot bind to the promoter and initiate transcription
TRANSCRIPTION ON:
-Inducer binds to the activator and CHANGES its shape, enabling it to bind to the site.
-RNA polymerase can then BIND to the promoter and initiate transcription
During Lac operon, what happens in transcription when glucose is present and lactose is present?
Transcription not activated. Low cAMP-, CAP cannot bind. Inducer(allolactose) prevents repressor from binding to the operator.
During Lac Operon, what happens in transcription when there is no glucose and no lactose?
Transcription activated, but blocked. High cAMP-, CAP/cAMP complex binds to activator-binding site. Repressor is bound to operator, however, blocking polymerase
What does the Lac Operon control?
controls the expression of genes involved in lactose catabolism
e.g:
-lac Z gene: B-galactosidase
-lac Y gene: Permease
-lac A gene: Thiogalactoside Transacetylase
During Lac Operon, what happens when there is no glucose, but lactose is present
Transciption activated. High cAMP-; CAP/cAMP complex binds to activator-binding site. In addition, inducer (allolactose) prevents repressor from binding to the operator
What is the action of Permease? B-galactosidase?
Permease: actively transports lactose into the cell
B:galactosidase: breaks down the lactose into galactose and glucose
During Lac Operon, what does glucose do?
When glucose is present, glycolysis is not operating, so there isn’t cAMP. cAMP needs to be present to bind to CAP to start transcription. cAMP and CAP need to be binded together to attach RNA polymerase to the promoter.
_________ growth curve occurs in the presence of glucose and lactose.
*BE ABLE TO DRAW THIS AND EXPLAIN IT!
Diauxic
-two growth periods, one on glucose and one on lactose.
-LAG, LOG GROWTH (on glucose), STATIONARY, LOG GROWTH (on lactose), STATIONARY
During Lac Operon, what does lactose do?
Without lactose, the repressor is binded to the operator site, blocking transcription. Lactose needs to be present to have the inducer release the repressor.
Lac Operon is also controlled by Catabolite Repression.
What is Catabolite Repression?
When glucose is LOW, cAMP is HIGH, binds to Catabolite Activator Protein (CAP) to induce transcription
In presence of lactose, allolactose is produced, BINDS to repressor, FALLS OFF operator so that transcription can proceed
ESSAY QUESTION
What happens in the following situation of a Lac Operon:
- No glucose, no lactose
Transcription ACTIVATED, but BLOCKED
-High cAMP
-CAP/cAMP complex binds to the activator-binding site, but a repressor is bound to the operator, blocking RNA polymerase
ESSAY QUESTION
What happens in the following situation of a Lac Operon:
- Glucose present, no lactose
Transcription NOT ACTIVATED and BLOCKED
-Low cAMP
-CAP CANNOT bind, and the repressor is bound to the operator, blocking RNA polymerase
ESSAY QUESTION
What happens in the following situation of a Lac Operon:
- Glucose present, lactose present
Transcription NOT ACTIVATED
-Low cAMP
-CAP CANNOT bind. At the same time, the inducer (allolactose) prevents the repressor from binding to the operator
ESSAY QUESTION
What happens in the following situation of a Lac Operon:
- No glucose, lactose present
Transcription ACTIVATED
-High cAMP
-CAP/cAMP complex binds to the activator-binding site and the inducer (allolactose) prevents the repressor from binding to the operator
what is the purpose of the trp operon
the purpose of the trp operon is to make enzymes needed in the biosynthetic pathway for making tryptophan
what happens if there is plenty of trp present
if there is plenty of trp is present, the trp will serve as a co-repressor, activating the repressor to stop transcription
what does UTR mean?
untranslated region
trp is a repressible operon.
What happens when tryptophan is LOW?
- The trp repressor is normally inactive
- It cannot bind to the operator
- So, transcription takes place
trp is a repressible operon.
What happens when tryptophan is HIGH?
- Tryptophan binds to the repressor and makes it active
- The trp repressor binds to the operator and shuts transcription off
What are the TWO roles of the hairpin loops in the trp operon?
WHEN TRP IS HIGH:
-Hairpin prevents transcription; produces the attenuator that terminates transcription
WHEN TRP IS LOW:
-Hairpin allows transcription; attenuator never forms, thus allowing transcription to continue
A regulator gene produces a repressor in an inducible operon. A geneticist isolates several constitutive mutations affecting this operon (meaning constitutive expression of operon).
- Where might these constitutive mutations occur?
- How would the mutations cause the operon to be constitutive?
- the repressor is active and binds to the operator, inhibiting transcription. For transcription to occur, an inducer must bind to the repressor making it unable to bind the operator.
- Constitutive mutations might occur in the regulator gene, altering the repressor so it can’t bind to the operator. Alternatively, constitutive mutations might occur in the operator so that the repressor cannot bind.
T/F: One mutation has a greater effect in a haploid organism than in a diploid organism
TRUE
What is the difference between a Wild-Type and a Mutant?
Wild-Type: organism normally found in nature
Mutant: an organism that has an acquired changed from mutation
What is an Auxotroph?
organisms that NEED added nutrients
What is a Spontaneous Mutation?
mutation that occurs randomly, but at a predictable rate
can be spontaneous from errors in replication, repair, and recombination (movement of large sections of DNA)
Mutants will rarely emerge as a significant part of the population.
What is a time they are?
when they are selected from (e.g., in the presence of an antibiotic)
What are the three types of methods for selecting Mutants?
- Direct Selection
- Indirect Selection
- Penicillin Enrichment
What is Direct Selection?
for selection of antibiotic-resistant mutants
Placed on an agar plate containing an antibiotic and one without (e.g., Transformation lab)
What is Indirect Selection?
for auxotrophic mutants
uses cotton velvet as a stamp; an enriched complex medium and synthetic medium is used
Enriched complex medium = ALL colonies grow
Synthetic medium = auxotrophs DO NOT grow
What is Penicillin Enrichment?
selects for auxotrophic mutants (another way)
Penicillin is added to synthetic medium, which kills actively multiplying cells
Most prototrophs are KILLED; auxotrophs SURVIVE because they cannot multiply in the medium
What is Transformation in bacteria?
transfer of “naked” DNA
What is Transduction in bacteria?
transfer of DNA via bacteriophage
What is Conjugation in bacteria?
transfer of DNA via direct contact between two bacterial cells
List the steps that occur in DNA-mediated Transformation.
- Transforming DNA (double-stranded) attaches to the recipient cell surface
- Single strand of donor DNA enters
- Pairing of donor DNA with homologous region of recipient chromosome
- Integration of single-stranded donor DNA by breakage and reunion
- Transformed cells multiply; non-transformed cells die when placed on antibiotic medium
What does it mean when a cell is “competent?”
refers to its ability to take up foreign DNA from the environment
Can you induce competence?
Yes - depending on the bacteria it must be in a particular part of the growth curve or a special physiological condition
Means such as electroporation is a way in which to allow DNA into a cell, thus making it competent
What is Electroporation?
an electric field generates pores in bacterial membranes allowing the entry of the exogenous DNA molecule
Explain Transduction
Transfer of DNA from one cell to another through a replicating virus (known as phages)
- Phage enzymes degrade the bacterial DNA
- Phage genome now controls the cell’s replication
-Make new viral particles
-Pack bacterial DNA into a phage protein coat - Cell lyses
- Release of bacterial phage.
- Recipient incorporates donated DNA by recombination.
What are the steps in which Conjugation is able to occur between two bacteria cells?
- The donor must have the gene for pili or sex pili, which is located on the F plasmid (i.e., they are F+)
- Donor cell (F+) connects to the recipient (F-) with its pilus
- Pilus draws cells together
- One strand of the F pilus transfers to the Recipient
- The Recipient now synthesizes the complementary strand of the F plasmid to become an F+ cell with a pilus
- The Donor synthesizes a complementary strand restoring its plasmid.
What is the difference between the following:
- F’
- F-
- HFR
- H+
- F’ = plasmid has come back out of HFR
- F- = no plasmid (recipient cells)
- HFR = plasmid has incorporated into chromosome
- H+ = has plasmid
What are 2 virulence factors found on plasmids?
-Adhesions
-Enterotoxin
If 75% of one E. coli strain’s genes exist in all strains (the “core genome”), where is the remainder located?
made up of mobile genetic elements such as plasmids, transposons, bacteriophages, and genomic islands
What are Transposable Elements?
Segments of DNA that are 700 - 40,000 bp in length that move (transpose) from one location in a DNA molecule to another.
Often causes mutation if the gene is interrupted
What do Transposons contain?
Transposons all contain plaindromes
e.g., GAATTC is the palindrome of CTTAAG
they vary in their sequence in genetics, this is called an inverted repeat