Cellular Control + Genome manipulation Flashcards
what is gene expression ?
gene is transcribed and used to make a functional protein (switched on)
which stages is gene expression controlled ?
transcriptional, post-transcriptional, post-translational
what are transcription factors ?
proteins that bind to DNA and switch on/off genes by increasing/decreasing rate of transcription
state the name of transcription factors that switch genes on
activators
state the name of transcription factors that switch genes
repressors
how is gene expression controlled at transcriptional level?
shape of transcription factor altered (caused by certain molecules) – changes binding ability – changes rate of transcription
where do transcription factors bind in eukaryotes ?
specific DNA sites near start of target gene
where do transcription factors bind in prokaryotes ?
operon
what is an operon ?
section of DNA containing structural genes, control elements and (sometimes) regulatory gene
what are the structural genes in an operon ?
gene that codes for useful proteins (enzymes..)
what are the control elements in an operon ?
promoter - DNA seq. before structural genes that RNA pol. binds to
operator - DNA seq. that transcription factors bind to
what is the regulatory gene in an operon ?
codes for activator or repressor
what is the lac operon ?
E. coli gene that produces enzymes to respire lactose when not enough glucose
what are the three structural genes of the lac operon ?
lacZ / lacY / lacA
Describe lac operon when no lactose present
- regulator gene produces lac repressor
- lac rep. binds to operator site
- blocks RNA pol. from binding to promoter
Describe lac operon when lactose present
- lactose binds to repressor
- repressor changes shape so can’t bind to operator site
- RNA pol. can transcribe structural genes
what are introns ?
DNA sections that don’t code for amino acids
what are extrons ?
DNA sections that do code for amino acids
what happens at post-transcriptional level?
pre-mRNA is spliced to form mature mRNA
what is splicing ?
removal of introns
what happens at post-translational level?
cAMP produced from binding of molecules to membrane – cAMP activates protein by changing 3D structure
describe how cAMP activates Protein kinase A (PKA)
- cAMP changes 3D structure
- causes subunits to unbind from one another
- PKA now active
what are hox genes ?
genes which code for proteins that control body plan development
what are homeobox sequences ?
highly conserved (little change between organisms) regions of hox genes
How do hox genes control development?
- homeobox seq. codes for homeodomain
- homeodomain binds to DNA
- protein enabled to work as transcription factor
what is apoptosis ?
programmed cell death
describe apoptosis process
- enzymes break down components
- cell shrinks and breaks into fragments
- fragments are engulfed and digested by phagocytes
give example of how gene regulates apoptosis from internal stimuli
DNA damage can trigger apoptosis
give example of how gene regulates apoptosis from external stimuli
attack by pathogen
what are the three terms for changing DNA seq. ?
substitution / deletion / insertion
state the three ways a mutation can be neutral
- mutation doesn’t change amino acid
- mutation changes amino acid, but is chemically similar so functions similarly
- codes amino acid not involved w/ function
what does PCR do?
amplify fragment of DNA
what does PCR stand for ?
polymerase chain reaction
what are the reactants for PCR?
DNA sample, free nucleotides, primers, DNA polymerase
what are primers?
short pieces of DNA complementary to start of DNA fragment
outline the process of PCR (3)
- heat mixture - break H bonds
- cool mixture - primers anneal
- heat a bit - DNA pol lines up free nucleotides (com base pair)
Two copies of DNA formed