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Biochem 275: Exam 1

Griffith, 1928 Rough & Smooth Bacteria
Rough wasn't harmful while smooth killed mice both heated -> mice survived heat R, not S -> mouse died heat S, not R -> mouse still died and S was present
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Avery, 1944 DNA vs Protein
broke up the smooth bacteria into protein and DNA DNA was the genetic material because when put into the R bacteria it killed the mouse
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Hershey, 1952 Bacteriophage
Bacteriohage head contains the DNA Radioactive phosphorus and sulfur to determine that DNA was the genetic material
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Nucleotide
Deoxyribose + base + phosphoric acid
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Nucleoside
Deoxyribose + base
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Linus Pauling, 1951
First helical structure
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Franklin and Gosling, 1951 XRay Diffraction
concluded there are about 10 base pairs per 360 degrees
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Chargaff's Rule
1:1 Ratio of purines:pyramidines
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Forces within DNA in increasing order
Dispersion, Hydrogen, Ionic, Covalent
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G-C bonds vs A-T bonds
The G-C bonds are about 3 kcals stronger than the A-T bond
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Restriction Enzymes
Like ecoRI, cut DNA after recognizing specific sequences
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Base + deoxyribose
-osin EX: adenosine
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Base + deoxyribose + 1 phosphate
base-osine monophosphate Ex: adenosine monophosphate
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Base + deoxyribose + 2 phosphates
adenosine diphosphate (ADP)
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Base + deoxyribose + 3 phosphates
Adenosine Triphosphate (ATP)
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ATP, TTP, GTP, CTP
Building blocks because they link together for the backbone. Forms 2 phosphate groups
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Type I Restriction Enzymes
cut at random positions far fro the recognition sequences so they're not used in labs
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Type II Restriction Enzymes
cut at defined points close to or within the recognition sequences (often used in labs)
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How do bacteria protect their DNA?
Methylated DNA cannot be recognized or cleaved by the restriction enzymes like ecoRI
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Where does cleavage occur in DNA?
Phosphate backbone and it regenerated a hydroxyl group
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Sticky Ends: Blunt Ends
cut in the same spot on both sides and can be ligased back with many other sequences
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Sticky Ends: Cliffs
strands are uneven and cut at same base resulting in pre-determined binding to specific strands
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Features of Bacterial Plasmid (2)
origin of replication that is <100 bps Promote genes of antibiotic resistance
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β-lactam
Antibiotic resistant gene
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Cloning
Cleaving DNA and using DNA ligase to add in an antibiotic gene, now some DNA has the resistance
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Gel Electrophoresis
Agarose gel where smaller strands go farther towards positive charge. Mix DNA with ethidium bromide to make it visible
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mRNA
Messenger intermediate between DNA and protein
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tRNA
Transfer bind to amino acids and mRNA to link protein anticodon on one end and a protein on the other to bind together to make polypeptide chains of amino acids
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rRNA
Ribosomal Interacts with proteins
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miRNA
Micro (20 bps) Tandemly repeated DNA
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snoRNA
Small Nucleolar (Guide) Modifies mRNA
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snRNA
Small Nuclear Helps proteins cut RNA and put it together
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RNA vs DNA (3 differences)
RNA uses ribose not (deoxy) uracil not thymine RNA is single stranded and can fold on itself
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AU Bond
Doesn't form a major/minor groove 2 Hydrogen bonds are formed
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UAU Triplet
now possible because RNA strand can twist
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Genome
All DNA (bases)
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Isoforms
Genes encoding for multiple, similar proteins
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Gene
DNA sequences coding for 1 protein that influences a trait
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Chromatin
DNA an its associated proteins
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Compactors of DNA
Histones
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Find binding sites and bring in assembly team
transcription factors
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Protect DNA from degredation
DNAse
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Other proteins in Chromatin
allow DNA to be separated and proteins that keep pairs together
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Nucleosome
DNA bound to histones
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Gene Density
Decreases as the complexity increases a lot
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Noncoding DNA in bacteria
Regulatory sequence that determines expression
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Noncoding sequences in prokaryotes
introns
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What do we achieve by coding from both 5' to 3' and 3' to 5'?
There can be many genes overlapping coding for different things
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Base and Ribose modifications result in...
New functionality
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Pseudouridylation
uracil mirrors and spins to create a new base pair bond
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2'-O-Methylation
adds a CH3 to where the H was on the 2' position
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Secondary Structure of RNA
The folding allows for important purposes
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10 nm fibers
DNA is wrapped around histones
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30 nm fibers
DNA is wrapped around histones and then this refolds onto more histones
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Core DNA
is wrapped around core histones and is 147 base pairs long
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Linker DNA
is between histones, is 20-60 base pairs long and allows DNA to fold into 30 nm fibers
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The Positive Amino Acids
lysine and argenine
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Dimers
H2A and H2B
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Tetramers
H3 and H4 that are attracted to DNA's negative charge
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Intermediate Stage
DNA wrapped around tetramer
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Final Stage of DNA Packing
Two dimers add onto the tetramer and DNA for 8 protein
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N-Tails
N-Tails are hanging off (as proved by use of protease) Tails can be methylated or acetylated through these tails, modifying the hisotnes
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