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Purple, brown, white, pink

Independent var- what I manipulate, only 1


Dependent- what changes in reaction
Observ, Hyp, Pred(about dep var.)
Negative cont.- know effect of dep. Var w/out indep.
Positive- known effect of dep. Var.
Experimental: variations in indep.; unknown effect on dep.
C1 x V1 = C2 x V2 where
1M = 1 molar = 1,000 mM = 1,000,000 M 1mM = 1 millimolar = 1,000
M 1M = 1 micromolar
1L = 1 liter = 1,000 ml = 1,000,000 l.. 1ml = 1milliliter = 1,000 l..
1l = 1 microliter
DNAtranscrRNAtranslProtein
Restriction sites allow cloning of foreign DNA
Enzyme is a protein, not all proteins are enzymes
-We Transformed E. coli cells with pBR322 gene which contains the gene for
-lactamase
- -lactamase causes cells to be resistant to ampicillin
Corkscrews (Spirilla), Rods (Bacilli), Dots (Cocci)
Peptidoglycan (cell wall protects) made from carbohydrate strands & peptide
cross-links
Periplasmic Space: btwn the inner and outer membrane in gram-negative
bacteria
Plasmid: A circular piece of DNA that often contains antibiotic resistance
genes
-lactamase (periplasmic): enzyme breaks down penicillin like molecules
into products
Gram positive- single cell membrane that is surrounded by a thick cell wall.
Gram negative- more resistant bc they have a double membrane, an inner
and an outer.
Gram Stain Order- crystal violet, iodine, alcohol, safranin
Positive control: Stain known Gram positive/negative bacteria.
Negative control: Examine unstained bacteria.
Controlled Variables: (place two knowns with an unknown on each slide)
Substrate/Reactants = what goes in, Products = what comes out
G = Reaction Energy = energy taken to get from A (reactants) to B
(products)
Ea = Activation Energy = energy taken to get from A (reactants) to transition
state

-Enzymes LOWER Ea so that a reaction can occur & TS can be reached,


G is unaltered
Isotonic Solution - Both solutions have identical (the same) concentrations
of solute.
Hypertonic Solution - Concentration of salt is lower & water is higher inside
the cell than outside the cell. Water move out of cell
Hypotonic Solution - Concentration of salt is higher and water
concentration is lower inside the cell than outside. Water move in cell
Suicide inhibition- Cell wall synthesizing enzymes inhibited by penicillin.
Penicillin forms covalent bond at active site of enzymes.
Penicillin binds at the active site of CWSE.
Penicillin is a substrate and not an inhibitor of beta-lactamase.
Beta-lactamase converts penicillin into a biologically inactive product.
osmotic shock- process by which we lyse the beta lactamase
Negative Control: Readings of nitrocefin without -lactamase.
Positive Control: Use already isolated/purified -lactamase.
-Nitrocefin is a substrate of -lactamase, yellow, broken down turns red
--lactamase does not change during this reaction.
Solute - substance that is dissolved or put into the solvent (Salt, Sucrose)
Solvent - the liquid into which the solute is poured and dissolved (Water)
Diffusion: random movement of molecules due to ke, from high
concentration to low
Osmosis: the diffusion of water molecules through a semi-permeable
membrane.
Vmax: maximum rate at which a reaction will proceed
Km: the concentration of substrate that yields 1/2 Vmax
-Measure of binding affinity, measured in substrate concentration units
- Higher Km= lower affinity for the substrate; takes more substrate 2 reach
same velocity
-Lower Km = higher affinity for the substrate; takes less substrate to reach
same velocity
Substrate concentration- x axis Rate of product formation- y axis
Competitive Inhibition
Inhibitor only binds to the free enzyme, prevents substrate from binding.
- often binds in the active site, not always
- usually structurally similar to the substrate, not always
-Vmax is unchanged: unaltered
-Km is increased: higher substrate concentrations required get same
velocity of Vmax
Uncompetitive Inhibition

-Addition of substrate actually makes the inhibitor bind better!


-Km and Vmax are both decreased by the same proportion.
-Vmax / Km ratio remains constant.
Noncompetitive inhibition
-Km may increase, decrease, or (rarely) remain the same.
-Vmax, however, always decreases, as with any inhibitor
CAN1 encodes for arginine permease (an enzyme)
Transports ARG normally, but can transport the toxic analog,
canavanine (CAN), if its available
CAN competes with ARG during translation to be incorporated
into proteins and makes them non- functional
CAN1 cells die on plates containing CAN (canavanine), because
functional
CAN1 (arginine permease) brings the toxin into the cell
Can-Sensitive (CanS)
can1 cells can grow on plates containing CAN (canavanine),
because the
transporter (arginine permease) does not work no toxin enters the cell
Can-Resistant (CanR)
ade1 and ade2 mutants turn the cells RED
Mutants before AIR (ade4-ade8) turn the cells WHITE
PRPP (1st compound in the pathway) is the substrate
AMP (last compound in the pathway) is the product