Catalytic Efficiency
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catalytically perfect - depend on rate of diffusion
,Gluconeogensis differences from glycolysis
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pyruvate (pyruvate carboxylase)
Oxaloacetate (phosphoenolpyruvate carboxykinase)
F1,6BP (fructose 1,6 bisphosphatase)
F6P
G6P (glucose 6 phosphatase)
Irreversible inhibition
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inhibitor binds so tightly therefore inactivates AKA suicide
IP
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Immunoprecipitation with proteins
1. add antibodies specific to protein of interest to lysed cells
2. add antibody binding protein on beads
3. centrifuge and protein is on bead with antibodies
4. gel elect
2,3 BPG
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allosteric effector for binding hemoglobin - blocks O2 from binding -
keeps in T state
Protein Separation Techniques (ALL)
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Homogenization
Salting Out
Affinity Chromatgraphy
Ion Exchange
Size Exclusion/Gel Filtration
H, S, A, I, SG
He said all individuals SinG
Western Blotting
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run gel - transfer to sheet - stained with radioactive antibody -
autoradiogram - exposed antibody
Vectors into plasmids
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, phage easily infects;
recombinant DNA
Lysogenic - DNA inserts into bacterial genome
Lysic - cells lysed as more phage are made
Protein Separation Techniques (A)
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Affinity chromatography
add a tag; histidine binds; see how well there is an affinity for the group
Restriction enzymes
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manipulate DNA, site specific endonucleases, cleaves bases and DNA
recognition sites are a palindrome
Cut into sticky or blunt ends
DNA polymerase vs. RNA Polymerase
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High fidelity vs. low fidelity
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catalytically perfect - depend on rate of diffusion
,Gluconeogensis differences from glycolysis
Give this one a try later!
pyruvate (pyruvate carboxylase)
Oxaloacetate (phosphoenolpyruvate carboxykinase)
F1,6BP (fructose 1,6 bisphosphatase)
F6P
G6P (glucose 6 phosphatase)
Irreversible inhibition
Give this one a try later!
inhibitor binds so tightly therefore inactivates AKA suicide
IP
Give this one a try later!
Immunoprecipitation with proteins
1. add antibodies specific to protein of interest to lysed cells
2. add antibody binding protein on beads
3. centrifuge and protein is on bead with antibodies
4. gel elect
2,3 BPG
,Give this one a try later!
allosteric effector for binding hemoglobin - blocks O2 from binding -
keeps in T state
Protein Separation Techniques (ALL)
Give this one a try later!
Homogenization
Salting Out
Affinity Chromatgraphy
Ion Exchange
Size Exclusion/Gel Filtration
H, S, A, I, SG
He said all individuals SinG
Western Blotting
Give this one a try later!
run gel - transfer to sheet - stained with radioactive antibody -
autoradiogram - exposed antibody
Vectors into plasmids
Give this one a try later!
, phage easily infects;
recombinant DNA
Lysogenic - DNA inserts into bacterial genome
Lysic - cells lysed as more phage are made
Protein Separation Techniques (A)
Give this one a try later!
Affinity chromatography
add a tag; histidine binds; see how well there is an affinity for the group
Restriction enzymes
Give this one a try later!
manipulate DNA, site specific endonucleases, cleaves bases and DNA
recognition sites are a palindrome
Cut into sticky or blunt ends
DNA polymerase vs. RNA Polymerase
Give this one a try later!
High fidelity vs. low fidelity
