Water, PH and buffer Specific properties of water Hydrogen binding, Acidity, H +, PH and buffer.
Physical Properties Due to the polar nature of water Hydrogen bond- weak attraction between hydrogen...
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Transcript of Physical Properties Due to the polar nature of water Hydrogen bond- weak attraction between hydrogen...
![Page 1: Physical Properties Due to the polar nature of water Hydrogen bond- weak attraction between hydrogen on adjacent molecules such as water H O H O Hydrogen.](https://reader035.fdocuments.net/reader035/viewer/2022062421/56649e495503460f94b3d449/html5/thumbnails/1.jpg)
Physical Properties
• Due to the polar nature of water
• Hydrogen bond- weak attraction between hydrogen on adjacent molecules such as water
H H O
H H
OHydrogen bond
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Water and it’s importance to Life
• Life evolved in water
• Water’s unique properties have made life as we know it possible
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Physical Properties• Heat of vaporization- amount of
energy that is released or gained when changing state from liquid to gas or back
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Physical Properties• High Specific Heat- the amount of
heat absorbed or released when water changes temperature by one degree C. ( 1 cal. )
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Ice Floats
• As a liquid water’s hydrogen bonds continuously break and reform
• As a solid four molecules form hydrogen bonds creating crystals with open channels and thus fewer molecules per area.
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Physical Properties• Water reaches maximum density
at 4 degrees C.
• Water is a universal solvent due to it’s polar nature
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Evaluate the importance of the following and explain the
property of water responsible.• Cytoplasm is 98 % water
• Ice Floats
• Lake effect temperature moderation
• Evaporative Cooling
• Spring-Fall Overturn
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Most Abundant Chemicals in Life
• Carbon
• Oxygen
• Hydrogen
• Nitrogen
• Ca, P, K, S, Na, Cl, Mg > 4 %
96 %
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Carbon is special
• Tetrahedral structure- four valence electrons shared
• Covalent bonds - stability
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Carbon is Special
• Variations are possible in carbon molecules that provide diversity
• Isomers are possible structural- differ in structure
same chemical formula geometric-differ in spatial
relationship enantiomers-mirror images of
each other
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Condensation Synthesis
A B+ A B + H2O
A and B could be monosaccharides or amino acids
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Hydrolysis
+ H2O +
Addition of water breaks the bond
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Polymers
Polymers are repeating units of monomers. They are very important to Biology.
They are made or synthesized by the removal of water called CONDENSATION SYNTHESIS
They are broken down by the addition of water orHYDROLYSIS
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Classes of Biomolecules
• Carbohydrates- used for energy and structures( building living organisms)
• Lipids- used for energy storage, communication and structures
• Proteins- used for a variety of life functions
• Nucleic Acids-the instructions for building life
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Carbohydrates
• Three common forms– Monosaccharides– Disaccharides– Polysaccharides
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Carbohydrates
• Monosaccharides- single sugars or simple sugars,ex. Glucose ( C6H12O6)
• Disaccharides- double sugar, ex. Sucrose
• Polysaccharides- polymers of glucose such as: 1. Starch 2. Cellulose 3. Glycogen 4. Chitin
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Review
• What will happen here?
AOH + HB = ?
And here:CH2OH CH2OH
OH
H
OH
OH
OH
OH
OH
OH
OH
O OH
H2O
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Dehydration Synthesisor a Condensation Reaction
A + B = AB + H2O
CH2OH CH2OH
OH
OH
OH
OH
OH
OH
O OO
H2O
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Review
• What will happen here?
AB + H2O = ?
And here:CH2OH CH2OH
OH
OH
OH
OH
OH
OH
O OH2O
O
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Hydrolysis or Reaction
AB + H2O = AOH + HB
CH2OH CH2OH
OH
OH
OH
OH
OH
OH
O OOH
OH
Molecules have been HYDROLIZED!
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GlucoseCH2OH
OH
H
OH
O
C
OH
OH
H
H H
H
Glucose has a chemical formulaof C6H12O6
CC
C
C
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O
C
CC
C
CH2OH CH2OH
FRUCTOSE
HO H
OHOH
HH
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Disaccharides
• Sucrose and Lactose
• 2 monosaccharides bonded together
CH2OH CH2OH
OH
OH
OH
OH
OH
O OO OH
Alpha or Beta?
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Polysaccharides
• 3 or more Monosaccharides bonded togetherCH2OH CH2OH
OH
OH
OH
OH
OH
O OO
OH
OH
OH
CH2OH
O
O
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Polysaccharide
• Starch-storage in plants
• Cellulose-structural part of plant cell wall
• Glycogen- storage in animals, liver
• Chitin – structural component for arthropods, exoskeleton. Also found in fungi.
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Polysaccharides – Starch
• Plants use it as energy storage
• Difficult for humans to break down– Ex. Avoid a high starch diet
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polysaccharides
Glucosemonomers
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Polysaccharides – Cellulose(B 1, 4 linkage)
• Long fibers• Up to 15,000 Glucose units per strand• Most abundant biological substance on
earth– Ex. Cotton, Trees, Paper
• Why is cellulose so strong?• Why can’t humans breakdown cellulose
and cows can?
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Polysaccharides – Glycogen
• Animals use it as energy storage
• Lots and lots of it in the liver
• Forms huge branched storage units which allow for easy break down for energy
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Other polysaccharides
• Chitin– Found in the exoskeleton of insects, and
arthropods• Ex. Crabs, lobsters, grasshoppers
• Pectin– Found in plant cell walls– Provides rigidity
• Heteropolymers– Glycoproteins and peptidoglycans
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ProteinPolymers of amino acids
With 20 natural amino acids thereare a variety of proteins
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Amino AcidsThe building blocks of protein
N -C - CH
H
O
OH
H
R
R- there are twenty different R groups possible
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Alanine
NH2-CH-COOH
CH3
Glycine
NH2-CH2-COOH
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Peptide bond- is a bond between amino acidsa molecule of water is removed
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Protein Structure
1. Primary- order of the amino acids2. Secondary- hydrogen bonds cause pleats and helix3. Tertiary- folds and loops create shape by R Group bonds4. Quaternary-interaction of several proteins
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A protein with secondary structure
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A protein with Tertiary Structure
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Lipids
• Large molecules that do NOT have an affinity for water; not soluble in.
• May have hydrophobic-water fearing and hydrophilic-water loving parts.
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Triglycerides
hydrophilic
hydrophobic
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Types of Lipids
• Made of hydrocarbons - • Triglycerides- fats, waxes, and
oils(saturated all single bonds C-C, unsaturated have double C=C bonds
• Phospholipids- attached phosphate replaces one of the hydrocarbon tails
• Steroids- Ring Forms of Hydrocarbons cholesterol and some hormones
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Triglycerides
• Saturated fats- single bonds make this a solid at room temperature and more difficult to digest.
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Unsatured Fats
• Triglycerides that contain double bonds ( dehydrogenated) are liquids at room temp and more digestable
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Nucleic Acids
• Made of monomers called nucleotides
• DNA- deoxyribonucleicacid
• RNA- ribonucleic acid
• These molecules carry all the hereditary information of living things
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DNA Basic Composition
• DNA is made up of nucleotides
• Nucleotides are made of
…………...Deoxyribose sugar
……………Phosphate
……………Base
bases are guanine,cytosine, thymine and adenine
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CATABOLISM
ANABOLISM
SYNTHESISRESPIRATION
ATP SYNTHESIS
FROM ADP + Pi
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Free Energy
• Ability to do work in the cell or ecosystem.
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Energy Transfer
• ATP formation
• + G
• ENDERGONIC
• Stores energy in phosphate bond
• ATP breakdown
• - G• EXERGONIC
• Releases energy between phosphates
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Enzyme Characteristics
• Lower the activation energy
• Speed up the rate of a reaction
• Act as catalysts• Are proteins
(occasionally RNA)
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Enzyme Characteristics
• Conformation or shape is most important feature ( Lock and Key Hypothesis)
• Substrate Specific
• Do NOT become part of reaction
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Enzym
e activity.
pH7
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Enzym
e activity.
pH7
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Enzym
e activity.
Temperature o C10
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Enzym
e activity.
Temperature o C10 30
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Allosteric site
Active site
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substrate
products
Enzyme
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Cofactors• Non protein helpers for enzyme activity
• May bind to active site tightly or loosely
• Many are inorganic such as zinc or iron
• If organic they are called Coenzymes
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Allosteric site
• Regulatory site other than the active site.
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Enzyme
substrate
Enzyme-Substrate Complex
Competitive Inhibitor
Noncompetitive Inhibitor
inhibitor
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Active conformation
Inactive form
activatorActive site
Allosteric Regulation
Allostericsite
inhibitor
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Feedback inhibition
• Product may cause negative feedback (act to inhibit, disrupt conformation)
• Reactants may cause positive feedback ( act to preserve enzyme conformation)
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Enzyme a Enzyme b Enzyme c
Endproduct
Initialsubstance
- feedback
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Prokaryotic Cells
• Lack a nucleus
• Lack membrane bound organells
• Include bacteria and other Monerans
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Eukaryotic Cells
• Have a nucleus
• Have membrane bound organells
• Plants, Animals, Fungi, and Protists have these cell types
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Organells Membrane Bound ( endomembrane)NucleusEndoplasmic reticulum (rough)Endoplasmic reticulum ( smooth )Golgi apparatusLysosomeVacuolesVesiclesPeroxisome ( single membrane) MitochoindriaChloroplasts
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Non membrane bound organellsNucleolusMicrotubulesMicrofilamentsCentriolesCiliaFlagella
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Nucleus
• Chromatin- DNA organized with protein (histone)
• Controls Protein Synthesis• Double Membrane with pores may be continious with ER
• Nucleolus- made of and synthesizes RNA
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Endoplasmic Reticulum
• Rough ER- contains ribosome for protein synthesis
• Smooth ER- lacks ribosomes, synthesis of lipids, metabolism of carbohydrates, detoxification of drugs and poisons
Muscle ER- calcium ion transfer
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ER and protein synthesis and Transport Vesicles
• Export Proteins – become enclsed in vesicle of the ER Pinch closed
Especially secretory proteins ( glycoproteins )
E R..
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Golgi Apparatus
• Manufacture, storage shipping , and packaging secretion products
• Cis
• Trans
• Vesicles
cis
trans
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Phospholipids are amphipathic - have both hydrophobicand hydrophilic portions
hydrophilic
hydrophobic
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Membrane Fluidity
• Unsaturated hydrocarbons in the phospholipids make it flow laterally
• Cholesterol maintains some rigidity at low temperatures and prevents too much fluidity at high
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Membrane proteins
• Integral-penetrate into or through the lipid bilayer
• used for transport
• Peripheral protein- attach to the surface of the lipid layer
• used for- receptors, recognition(carbohydrates attached)
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Permeablility
• What passes through easily
• Oxygen
• Carbon dioxide
• water
• What does not pass through easily
• ions
• proteins
• carbohydrates
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Transport
• Passive• Requires no
expenditure of ATP• Moves from high to
low• Can be facilitative-
aided by protein conformation change
• Gatted channels
• Active• Requires ATP energy • Generally moves
from low to high• Gatted channels• Na-K pump• Proton Pumps• Phagocytosis or
Pinocytosis
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Solutions
• Homogeneous-same throughout
• solvent- what you are dissolving into
• solute- what you dissolve
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solution
*
SOLVENT
SOLUTE
*
* *
*
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Hypertonic
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.5M glucose .5M glucoseDistilled water1.5 M glucose
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Water Balance
• Plasmolysis( plasmolyzed) plant cell shrinks or looses water
• Flacid -plant cell gains water and looses at same rate
• Turgid- Plant cell gains
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Plasmolysis
• Membrane shrinks due to water loss
• Restricted to cells with walls
• Occurs in a Hypertonic environment
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Facilitated Diffusion• Proteins make movement of polar
molecules, ions, or larger compounds possible by providing a passage.
• Often protein changes conformation
• NO ATP required
• Movement from high to low concentration
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Active Transport
• Sodium Potasium Pump
• Proton pumps
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Membrane Potential
• Voltage across a membrane
• -50 to-200 millivolts
• Electrochemical gradients- combination of ion potential and electric charge difference
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Gated Channels
• Chemical or electrical impulses cause them to change shape-OPEN
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Na-K Pump
• Membrane Potential-voltage difference across a membrane
• Chemical Gradient-difference in concentration of solute across a membrane
• Electro-chemical Gradient- combination of the above ex.
• Na+,K+, Cl-(8.13)
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Exocytosis and Endocytosis
• Phagocytosis- engulf pseudopodia
• Pinocytosis- gulps
• Receptor mediated pinocytosis
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Signal Transduction
• Binding of extracellular molecule to receptor protein see model on pg. 156 Campbell
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Cell types determine cycle
• Prokaryotes- binary fission circular chromosome attaches to inner membrane. Replication is followed by reattachment at two sites.
• Eukaryotes-have a larger genome and nuclear genetic material must be carried on several chromosomes by specialized structures.
No spindle fibers
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Can be Confusing?ChomosomesChromatidsSister chromatidsHomologous chromosomesCentromereCentrosomeCentriolesKinetochoreNonkinetochore
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Chromosomes
DNA is continuous and wound around protein which is coiledand super coiled into a dark staining body.
Chromosomes can be seen as having two arms and often one is longer.
When duplicated the chromosome has identical sister chromatids held together at the centromere.
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chromatid
centromereSister chromatid
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Chromosome Number is fixed in each species
Diploid Number Monoploid (haploid)
2n n
46 in humans 23 in humans
Somatic Cells-bodycells
Gametes-egg, sperm, pollen.
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Cell Cycle
Events in the growth, development andreproduction of the cell.
Go cells have stopped dividing or have lostthe potential to divide.
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G1- gap or growth after cell division. Cell grows in size. this stage contains the RESTRICTION point
S- synthesis of new DNA from existing template(replication)
G2- gap 2 or growth prior to cell division
M- mitosis or chromosome division
C- cytokinesis or cell division
Interphase= G1, S, and G2
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Control of Cell Cycle
Restriction point- go/no go control during G1
G0 - a non-dividing stage for a cell
Growth Factors-compounds which regulate growth and division. Ex.PDGF platelet derived growth factor
Density-dependent inhibition- crowding inhibits celldivision.Adhesiveness- cells ECM causes them to stick together
Metastasis-cells(cancerous) migrate
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Cell Clock Regulators
• Proteins ( enzymes) regulate cell cycle
• Produced by internal cell clock genes
• Protooncogenes- cause cells to divide
• Tumor suppressor genes- prevent cell division
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Cancer and the Cell Cycle
• Normal Cells
• Adhesive
• Contact inhibition
• Cancer cells
• Lost adhesiveness
• Lost contact inhibition
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Principles of Heredity• Alternative versions of genes (alleles)
account for variations in a trait.• For each character, an organism inherits
two alleles, one from each parent.• If alleles differ, then the dominant will be
fully expressed over the recessive.• The two alleles segregate (separate)
during gamete formation.• Alleles on different chromosomes
segregate independently of one another
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Crossing over
During prophase of meiosis homologous pairs may exchange
genetic material.
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New Genetic Combinations
• Recombination during fertilization brings together two sets of genetic instructions
• Meiosis-crossing over brings about new combinations
• Random genetic mutation can result in random genetic change
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Electron CarriersNAD+ nicotinamide adenine
dinucleotide
NAD+ When oxidized
NADH +2 H+ When reducedFAD or FADH2
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Types of Respiration
• Anaerobic-without oxygen 1. Alcoholic fermentaion 2.Acetic Acid fermentaion 3. Lactic Acid fermentaion
• Aerobic-with oxygen
• ALL OF THESE BEGIN WITH THE ANAEROBIC PROCESS OF GLYCOLYSIS
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GLYCOLYSIS
Glucose is made ready to metabolize by addition of phosphates and then it is broken down into 2- 3 carbon compounds (PYRUVATE)
This yields a net gain of 2 ATP
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ACETYL COA FRORMATION
• Pyruvate is converted into a 2 Carbon compound and added to an enzyme
• CO2 is released
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Kreb’s Cycle
• Breaks C-H-O bonds• Energy is transferred via carriers
to other steps• CO2 released• Some small amount of ATP is
produced
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Electron Transport
• Hydrogen Pathway- pumps H ions
• Electron Transfer
• Chemiosmosis- H ions flow through ATP syntase proteins to make ATP from ADP + P
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Substrate level phosphorylation
• ATP is formed as a direct transfer of
electrons from the substrate to as ADP + P ATP
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Oxidative phosphorylation
• Electrons made available in metabolism are transferred to oxygen and ATP is produced in the process. Chemiosmosis
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Fermentation generates ATP by substrate level phosphorylation.It is anaerobicThree Types:
Alcoholic-
Lactic Acid- 2NAD+2Lactate
2 Ethanol +2CO2+NAD
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Photosynthesis
CO2+H2O light CnH2n0n+O2
Light- measured as an absorption spectrum, the wavelengths that are most important are different for different types of autotrophs
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light
PS1
PS2
Calvin CycleCalvinCycle
ATP
NADPH
Photolysisand Photophosphorylation
O2 CnHnOn
CO2H2O
Thylakoid
Stroma
Light rxn. Dark rxn.
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Visible Spectrum
Absorbed Absorbed680-700
Reflected
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Chl a
Photosystem I P700
700nm LIGHT
Primary acceptor
pq
pc
Cytochromecomplex
Photosystem IIP680
NADP+ + 2H
NADPH
680nm
-e
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Photosystem I
• Also known as P700-receives electrons from those released in PSII to replace photoexcited electrons uses light at far end of the red wavelength
• PSI 700• PSII 680 the II in PSII H2O
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Photosystem II• P680• due to an association with different
proteins • this system utilizes different
wavelengths• causes water to split capturing it’s
electron• it then transfers the electron to
PSII chlorophyll molecules
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Noncyclic Electron FlowWater is split (photolysis) and electrons pass continuously from water to NADP+.Primary electron acceptors pass photoexcited electron to the electron transport chain(Pq), (Pc), cytochromes..
Generates O2, NADPH, and ATP
Uses both PSI and PSII
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Cyclic Electron Flow
Excited Electrons pass through the electron transport chain from P700 (PS I ) and return to the starting point.Uses only PSI
Only ATP is generated
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Calvin Cycle
6ATP
3ATP
3ADP
6ADP
6NADPH
6NADP
3CO2
G3P--GlucoseRegeneration RuBP
Carbon Fixation
rubiscoRuBP
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Photorespiration
CO2 can act as a limiting factor.In cases where there is not sufficient Carbon dioxide plants will combine oxygen with RuBP to form compoundsthat are broken down into CO2
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Adaptations for Photosynthesis
• C4 Plants• CO2 is added to PEP
phosphophenolpyruvate
• stored in BUNDLE SHEATH CELLS near veins of leaf
• example- Corn
• CAM plants
• in hot dry areas plants must close stomates
• CO2 taken in at night is stored as an acid
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Discovery of DNA1. Frederick Griffith
– Was studying Streptococcus Pneumonia– Smooth vs. Rough Strains– Smooth had a mucous coat and were
pathogenic (caused pneumonia)– Rough were non-pathogenic– Conducted an experiment with mice– Found out that the Rough bacteria
became transgenic
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Discovery of DNA2. Avery, McCarty and MacLeod
– What was the genetic material in Griffith’s experiment?
– Purified the heat–killed S-bacteria• Into DNA, RNA, and Protein
– Mixed each with the R cells to see which one transformed
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Discovery of DNA3. Hershey-Chase Experiment
– Studied viruses that infect bacterial cells– Bacteriophages– Protein or DNA responsible for take-charge
actions of the virus?– Tagged the Protein with radioactive S
• Why?
– Tagged the DNA with radioactive P• Why?
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The Structure of DNA:a double helix?
• Chargaff’s Nucleic Acid Ratios1. Measured the base compositions of
several species
2. Percentage of each base present• Human DNA
1. A = 30% and T = 29%
2. G = 20% and C = 19%
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• Rosalind Franklin and Maurice Wilkins use X-Ray diffraction to view structure
• Watson and Crick propose a double helix using their X-Ray pictures
The Structure of DNA:a double helix?
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DNA Double Helix
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DNA: Three Parts
• DNA is made up of nucleotides
• Nucleotides are made of– Deoxyribose sugar– Phosphate– Base
• Guanine, Cytosine, Thymine and Adenine
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DNA: The Deoxyribose Sugar
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DNA: The Phosphate
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DNA: The Nitrogenous Bases
• Purines• Adenine and Guanine • Double Ring Structure
• Pyrimidines• Thymine and Cytosine• Single Ring Structure
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Single Stranded DNA
Nucleotides can only be added to the 3’ endof the nucleotide and therefore addition ofnew nucleotides is always 5’-----> 3’
DNA is anti-parallel!!
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DNA STRUCTURE
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How does it know to pair up?
• ADENINE ALWAYS PAIRS WITH THYMINE
• Two hydrogen bonds
• GUANINE ALWAYS PAIRS WITH CYTOSINE
• Three hydrogen bonds
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Why do they pair up?
• Double helix had a uniform diameter
• Purine + Purine– = too wide
• Pyrimidine + Pyrimidine– = too narrow
• Purine + Pyrimidine– = fits the x-ray data
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One last look
Why does it twist?
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DNA Replication
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Meselson-Stahl demonstrate theSemiconservative Replication of DNA using radioactive nitrogen
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Why must DNA Replicate?
•Species Survival– DNA must replicate BEFORE cell
division•Synthesis during Interphase
•All genes must be present in the daughter cells
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Origins of Replication
• Sites along DNA that contain specific nucleotides are recognized by specific proteins that initiate process
• In eukaryotes there are hundreds of thousands of such points
• Form replication bubbles
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How does DNA Replicate?
• Hydrogen bonds break, forming bubbles• Enzymes unwind and unzip• Free nucleotides in the nucleus start
process of complementary base pairing• Nucleotides are fused together by DNA
Polymerase only 5’ to 3’• Results in two identical double helixes
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Replication Steps• DNA helicase enzymes open double
strand• DNA uncoils and unzips exposing the
DNA template • Primase adds a RNA primer as binding
proteins hold strands together• DNA polymerase attaches to template at
replication fork• Nucleoside triphosphates add bases
pairing A-T and G-C as new strand is added to a 3’ end, primer removed
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replication3
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How does DNA Replicate?
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How does DNA Replicate?
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Leading Strand is Continuous
• A single RNA primers initiates the addition of nucleotides to the 3’ end of the leading strand
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Lagging Strand
• Must wait for replication fork to open and then add primer
• Form Okazaki fragments
• RNA is removed only after addition of about 100 to 200 nucleotides
• Fragments are joined by a ligase enzyme ( DNA glue)
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DNA Replication
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The result
• DNA Replication results in TWO double helixes. DNA unwinds and unzips, and new daughter strands form, each complementary to an old parental strand.
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RNA - Structure• “Ribonucleic Acid” – different from
DNA• Always Single Stranded• Ribose Sugar Base Unit• Phosphate group (same in DNA)• Nitrogenous Bases
– Cytosine always pairs with Guanine– BUT! Adenine always pairs with
URACIL• (different in DNA!!!!!)
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Four kinds of RNA
• Ribosomal RNA
• Messenger RNA
• Transfer RNA
• snRNA ribozyme in spliceosome
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rRNA
• Ribosomal RNA or rRNA • represents about 70% of cellular RNA
• joins with Ribosomal proteins to make the cellular organelle: RIBOSOMES
• FUNCTION – As a manufactured ribosome, supplies a location for tRNA to join with mRNA to synthesize a protein
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mRNA2. Messenger RNA or mRNA
• represents about 10% of cellular RNA • contains the sequence of bases coding for
a particular amino acid sequence in a polypeptide chain
• removal of non-coding, internal sequences (introns)
• modification of the 5' base (cap)• addition of adenines to 3' end (poly A tail)
FUNCTION – reads the DNA code (base sequence) and becomes a copy that is read at the ribosome to make a protein
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hn RNA
• Pre-mRNA contains introns-non-coding regions as well as exons-coding regions
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Processing mRNA
• Deletion of introns
• Join exons
• Add cap ( GTP) and poly A tail
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RNA splicing
• Spliceosome-several
snurps• snRNPs small nuclear
Ribonucleoproteins-splicing enzyme( cuts and glues)
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Transcription• DNA unzips at the locus of the gene
being coded• mRNA makes a copy of the gene
• Then…• mRNA is enzymatically modified
– A cap and a tail are added
• it then leaves the nucleus and finds a ribosome (composed of rRNA and protein)
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tRNA• Transfer RNA or tRNA
• represents about 20% of cellular RNA • each tRNA molecule is specific for one
amino acid • there is an enzyme for each amino acid
which recognizes the amino acid and its specific tRNA and joins the two together
• the specific joining of tRNA to amino acid is the only place where the “genetic code" applies
FUNCTION – Pairs with Amino Acids and delivers them to ribosomes at the right time to synthesize a protein
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Protein Synthesis
• Why should cells do this?– Cells would not be able to grow and change without
proteins.– Proteins are found everywhere:
• As enzymes, cell membranes, muscles, heart, blood…
• What happens when proteins are not made correctly or not made at all?– Ex. Cystic Fibrosis
• What part of DNA holds the code for the protein?
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PROTEIN SYNTHESISEveryone is involved
• Transcription
DNA, mRNA
• Translation
mRNA, rRNA and tRNA
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Pre-Translation
• mRNA binds to the ribosome
• Meanwhile, tRNAs are attaching to their amino acids using tRNA Transferase
• Free tRNAs, with their amino acids attached, circulate in the cytoplasm and match up with the triplet codes in the mRNA
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TranslationInitiation• The first tRNA enters the ribosome at the A
site• The second tRNA enters the ribosome (at the
P site) and the amino acids are bonded together = PEPTIDE BOND
Elongation• Both tRNAs shift in one direction and make
room for the next tRNA to enter the ribosome• this pattern continues until the protein is
complete
Termination• Stop codon is read – UAA, UAG, UGA
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Quick DefinitionsA-site – aminoacyl-tRNA binding siteP-site – peptidyl-tRNA binding siteTriplet code – DNACodon – RNAAnti-codon - tRNAIntrons – removed from initial mRNAExons – bonded together to make the
finished mRNA product for translationPolyribosomes – more than one ribosome
reading the mRNA at one time
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Codons and anti-codonsTriplet code on DNA TACmRNA copies it: CODON AUGtRNA carries the ANTICODON UACThe Genetic code reads the codon
AUG, the amino acid: Methionine
• 45 different anti-codons exist• AUG is always the initiation codon• GTP supplies energy needed to synthesize
protein• initiator tRNA always carries Methionine first!• Initiation factors - proteins that bring all parts
together (mRNA, small subunit, large subunit, and tRNAs)
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Genetic Code
• Interprets what the DNA triplet code reads
• Is written in both DNA base language A, G, C, T or RNA base language A, G, C, U
• Determines the order for Amino Acids
• Is universal within all species
• Reads the same as the anti-codon (on tRNA) except T is now U
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Genetic Code
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Gene RegulationControl of gene expression occurs at four levels in
human cells:Transcription and posttranscription control
(nucleus)Translation and posttranslation control (cytoplasm)• Various cells express different genes• Genes can be turned on or off• Genes respond to activity outside of the cell• Control of transcription is most important
regulatory mechanism (binding factors and enhancers) Presence of TF determines specialization
• Some binding factors are sensitive to hormones
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DNA Technology
• Biotechnology or genetic engineering – the use of natural biological systems to produce a product desired by human beings
• Examples include:• Gene Cloning• DNA Amplification• Transgenic Organisms• Gene Therapy• Chromosome Mapping and Sequencing
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Gene Cloning
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Gene Cloning• Recombinant DNA – DNA from two different
sources (human and E. coli)• Plasmid – circular DNA used to transport the
gene into the organism• Enzymes needed – Restriction and Ligase• Host cell – usually bacteria, wall must
become competent in order for the bacteria to uptake the plasmid
• Restriction enzyme cleaves DNA and allows for DNA fragment to insert at the sticky ends
• Vector – method of transporting a gene (virus, plasmid)
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pVIB lux genes
2 genes to produce LUCIFERASE
Aldehyde (energy source) synthesis
several genes
Regulatory genes to turn of and on
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DNA Amplification
Polymerase Chain Reaction – PCR
• Used to make multiple copies of the same gene
• Copies can be examined to see if they match any other sources
• Prevents constant extraction from the organism and better results
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Other Technologies• Recombinant DNA - gene
splicing
• Transgenic organism- an organism that contains another organism’s DNA
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Transgenic Organisms
• Transgenic – possessing gene(s) from another organism
• Gene Pharming – Using transgenic farm animals to produce pharmaceuticals
ex. CF, cancer, blood clots
• Genetically altering crops to be resistant to insects and produce larger
http://biology.about.com/science/biology/gi/dynamic/offsite.htm?site=http://abcnews.go.com/sections/science/DailyNews/gmcorn%5Fbutterflies000821.html
• Suicide Genes• Insulin
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Gene Therapy
• Delivering the defective gene to the cells that need it to produce a protein
• Familial hypercholesteremia
• SCID – severe combined immunodeficiency syndrome (missing maturation enzyme for T and B cells)
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Chromosome Mapping
• 100,000 human genes
• RFLPs – Restriction Fragment Length Polymorphisms – used to probe a region of DNA – visible under a microscope
• Restriction enzymes – sequence AA
• Specific base digestion– CF LAB
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Human Genome Project
• HGP – due for completion in 2002
• Already sequenced the Fruit Fly and E. Coli
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Gene Therapy
• Delivering the defective gene to the cells that need it to produce a protein
• Cystic Fibrosis
• Vector – method of transporting a gene (virus, plasmid)
– Mechanical - usually a laboratory tool used (inoculating loop)
– Biological - part or whole of an organism (bacteria)
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Chromosome Mapping
• 30,000 human genes
• RFLPs – Restriction Fragment Length Polymorphisms – used to tag a region of DNA – visible under a microscope
• Restriction enzymes – sequence AA
• Specific base digestion
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Sanger Method of DNA Sequencing1. Heat DNA Strands until they separate2. Add nucleotides and DNA Polymerase3. Add Dedeoxynucleotides (A, T, G, and
C) at different time periods to stop replication
4. Place fragments in to Gel Electrophoresis
5. Allow to migrate and read the Base Sequence
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Electrophoresis
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Human Genome Project
• HGP – due for completion in 2002
• Already sequenced the Fruit Fly and E. Coli
• The ultimate goal of HGP is to associate human traits and inherited diseases with particular genes.
• It promises to revolutionize both therapeutic and preventive medicine techniques for many human diseases.
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Human Genome Project
• Genome - the complete collection of an organism's genetic material.
• The human genome is composed of an estimated 30,000
• A single human chromosome may contain more than 250 million DNA base pairs, and it is estimated that the entire human genome consists of about 3 billion base pairs.
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DNA Fingerprinting
• Treat suspects’ blood with the same restriction enzyme
• Place sample in Gel Electrophoresis• Allow samples to migrate• Compare the suspects with the blood
found at the crime scene• Used in Criminal Trials: OJ Simpson
– OJ – DNA was an exact match yet he was found not guilty?