ethan hack material Flashcards
when end is RNA constructed from
5’
which end of an amino acid is positively charged and which is negatively charged
amine group (NH2) is positvely charged, carboxyl group (COOH) is negatively charged
name the 3 stop codons
UAG UAA UGA
what part of tRNA binds to the AA and the codon
anticodon binds to mRNA’s codon, 3’ end binds to AA
what type of 2D and 3D structure does tRNA have
cloverleaf (2D) and L shaped (3D)
what is the name of the enzyme that bonds AA’s to tRNA
aminoacyl-tRNA synthase
name a modified base that may be on the third position of a tRNA’s anticodon allowing it to bind to more than one codon
inosine
what is the location of translation
ribosomes
what are ribosomes made up of
protein and RNA
name the 3 binding sites on a ribosome and which sub unit are they one
Polypeptide and Anticodon, exit
small subunit
describe the initiation part of translation
1) first aminoacyl-tRNA binds to the P site of the small sub unit
2) codon-anticodon binding occours
3) large subunit binds
4) second aminoacyl-tRNA binds to the A site
discuss polycistronic DNA
multiple genes transcribed in a single mRNA molecule, like is done in prokaryotes from an operon
discuss prokaryotic RNA polymerase’s 2 forms
core enzyme; 5 subunits
holo enzyme; 6 subunits; also has a sigma factor
only the holo enzyme form can recognise the promotor region
how does prokaryotic RNA polymerase know where transcription stops
a ‘transcription termination sequence’ exists; mRNA folds into a stemp-loop structure which destabilises the mRNA-DNA interaction causing RNA to dissociate from the gene
how do prokaryotic and eukaryotic ribosomes know which codon is the first amino acid to match to an AA
prokaryotes; the shine-delgarno sequence of mRNA (close to start codon) binds to ribosomal RNA and acts as the ribosome binding site
eukaryotic; the ribosome binds to the begininning of mRNA and moves to the first AUG of mRNA
what is the shine- dalgarno sequence
AGGAGG. a sequence on prokaryotic mRNA which binds to the ribosome
discuss lactose structure
disaccaride consisting of 1 glucose and 1 galactose
discuss what the genes lacI, lacZ, lacY, lacA and lacI are expressed as and the function of the expressed products
lacA; beta-galactosidase transetylase; unknown function
lacI; repressor protein, binds to the operator region and stops transcription when lactose is absent
lacZ; beta-galactosidase; hydrolyses lactose
lacY; lactose permease; transports lactose into the e coli cell
discuss the 4 domains (polypeptides) of a a repressor protein
n terminal; binds to the DNA (operator region)
hinge region; holds N terminal in correct orientation
allolactose binding region
C terminal; used for two repressors to interact to form a dimer, which is needed for the repressor to bind to the DNA
discuss how allolactose interacts with repressor protein domain
allolactose binds to the allolactose binding region, and causes a conformational shape change which moves the hinge region; N terminal doesnt bind to DNA
why does binding of the repressor binding stop RNA polymerase action
operator and promotor sites overlap; binding of repressor protein means RNA polymerase cant bind
how many operator regions exist per operon. discuss these operators
3; one original operator and 2 auxillary operators. the auxillary operators are downstream of lacZ
binidng of repressor to 2 operators decreases transcription much more than binding of one repressor
what does i- mean
mutated lacI gene; mutated repressor protein which cant bind to the operon; transcription os constitutive (constant)
is the lacI gene part of the lac operon
no
discuss Oc (lap operon)
operator region is mutated; repressor cant bind; constitutive
discuss evidence for the lac operon mechanism
name the 3 scientists who worked on this
Jacob, Lwoff and Monod
looked at mutants with altered gene expression
the way that mutated beta-galactosidase (lacY) is screened for is by giving colonies the arteficial substrate X-gal, which is converted to blue product. those colonies which arent blue have a mutated gene and should be studied
when do lac operon mutation diagrams become complicated
when the cells are engineered to be diploid, cis and trans acting mutations now exist. trans mutation effect expression of genes in other DNA molecules to the one where mutation occoured
why is the lac promotor a weak promotor;
what is the name of a mutated promotor which isint weak, and why isint it weak
the -10 and -35 sequences are non standard
PlacUV5; -10 and -35 sequences are close to the consensus sequence
discuss of the importance of glucose concentrations being low in terms of lac operon regulation
when glucose concentration is low, cAMP concentrations are high; cAMP activates a CAP protein which binds to DNA’s -35 region, bends DNA 90 degrees. RNA polymerase binds to CAP with one of its sub units; increased binding to promoter occours if CAP is present
catabolic operons are switched (ON/OFF) when metabolite is present
Biosynthetic operons are switched (ON/OFF) when supply of product is low
ON
ON
discuss what the trp operon has genes for.
discuss how the operon’s expression is switched off
tryptophan AA
repressor only works when high concentrations of tryptophan exists, which binds to the repressor and allows it to bind to the DNA
define quorum sensing
discuss the bobtail squid being bioluminescent
discuss pathogenic bacteria
regulating gene expression in response to cell population density
symbiotic bacteria in the squids light organ are responsible for the light
these cells synthesise AHL (luxI gene), which diffuses out of the cell
if the cell density of bacteria is high enough then the concentration of AHL incrases, bacteria take it up, it binds to the LuxR gene, which activates transcription of the lux operon; light emmision
when invading a host, bacteria only produce degradative enzymes if the density of cells is high enough