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transformation
a change in genotype andphenotype due to the assimilation of external DNA by a cell
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DNA
- a nitrogenous (nitrogen-containing)base, a pentose sugar called deoxyribose, and a phosphate group
- adenine (A) 2(no O), thymine(T)1(CH3), guanine (G)2(O), or cytosine (C) 1(N-H-H)
- sugar-phosphate backbones are antiparallel
- A, G = purines
- C,T = pyrimidine
- Pairing = purine + pyrimidine, A+T, C+G
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Bond
- Bond with hydrogen bonds wiht nitrogenous base
- A+T : 2 Hydrogen bonds
- G+C : 3 Hydrogen bonds
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Gene
Have unique a unique order or base sequence
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Chargaff’s rules:
- (1) thebase composition varies between species,
- (2) within aspecies, the number of A and T bases are equal and the numberof G and C bases are equal.
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Three alternative models of DNAreplication.
Conservative model - The two parental strands reassociate afteracting as templates for new strands,thus restoring the parental double helix.
Semiconservative model.The two strands of the parental moleculeseparate, and each function sas a template for synthesis of a new, complementary strand
Dispersive model. Each strand of both daughter molecules contains a mixture of old and newly synthesized DNA.
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Meselson and Stahl Experiment
- comfirmed semi-conservative model
- cultured E.coli for several generations in N15 and N14
- result : band of hybrid DNA
- result 2: both light and hybrid DNA
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origins of replication
- site where replication of a DNA molecule begins
- short stretches of DNA having a specific sequence of nucleotides
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Prokaryotic Replication
- Ex. E.coli and bacteria
- only one origin of replication is present
- The parental strands separate at the origin, forming a replication bubble with two forks. Replication proceeds in both directions until the forks meet on the other side, resulting in two daughter DNA molecules
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replication in a eukaryotic cell
- DNA replication begins when replication bubbles form at many sites along the giant DNA molecule
- The bubbles expand as replication proceeds in both directions.
- the bubbles fuse and synthesis of the daughter strands is complete
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replication fork
Y-shaped region where the parental strands of DNA are being unwound
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Helicases
enzymes that untwist the double helix at the replication forks
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single strand binding proteins
bind to the unpaired DNAstrands, keeping them from re-pairing.
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Topoisomerase
helps relieve this strain by breaking, swiveling, and rejoining DNA strands.
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primer
- added to nucleotides
- The initial nucleotide chain that is produced
- is synthesized by the enzyme primase
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Primase
starts a complementary RNA chain from a single RNA nucleotide
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DNA polymerases
- Enzymes that catalyze the synthesis of new DNA by adding nucleotides to a pre existing chain(primer)
- require a primer and a DNA template strand, along which complementary DNA nucleotides line up.
- bacteria: 500 nucleotides/sec
- human: 50 nucleotides/sec
DNA polymerases can add nucleotides only to the free 3' end of a primer or growing DNA strand, never to the 5' end
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dATP, nucleoside triphosphate
- supplies an adenine nucleotide to DNA,is the sugar component, which is deoxyribose in the building block of DNA
- used for DNA synthesis are chemicall y reactive, partly because their triphosphate tails have an unstable cluster of negative charge
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leading strand
- 3' End
- Only one primer is required for DNA pol III to synthesize the leading strand
- DNA pol III remains in the replication fork on that template strand and continuously adds nucleotides to the new complementary strand as the fork progresses ( same direciton as the fork) right
- elongates continuously
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lagging strand
- 5' end
- DNA pol III work in direction away from the replication ,right
- synthesized discontinuously, as a series of segments (Okazaki fragments)
- 1) Primase joins RNA nucleotides into a primer
- 2)DNA pol III adds DNA nucleotides to the primer, forming Okazaki fragment 1
- 3)After reaching the next RNA primer to the right, DNA pol III detach
- 4) DNA pol I replace RNA with DNA andding 3' to frag 2.
- 5)DNA ligase - connect the fragments
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mismatch repair
- other enzymes remove and replace incorrectly paired nucleotides that have resulted from replication errors
- changes inDNA are usually corrected before they become permanent changes—mutations—perpetuated through successive replications.
- DNA repair enzymes: 100 in E.coli, 130 in humans
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Nucleotide excision repair
- 1. enzymes detect and repair damaged DNA
- 2.A nuclease enzyme cuts the damaged
- 3.DNA polymerase fills in the missing nucleotides
- 4. DNA ligase seals the free end
Ex. thymine dimers ( cuase by ultraviolet)
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Telomeric DNA
- acts as a kind of buffer zone that protects the organism’s genes.
- proteins associated with telomeric DNA prevent the staggered ends of the daughter molecule
shortening telomeric can lead to self destruction of tumor cell
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telomerase
catalyzes the lengthening of telomeres in eukaryotic germ cells, thus restoring their original length and compensating for the shortening that occursduring DNA replication
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bacterial chromosome
circular DNA molecule that is associated with a small amount of protein
- qukaryotic chromosome: one linear DNA molecule associated with a large amount of protein
- E.coli has 4.6 million nucleotide pair/4400 gene
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histones
proteins responsible for the first level of DNA packing in chromatin
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nucleosome
- basic unit of DNA packing; the “string”between beads is called linker DNA
- contains 4 histones
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nucleoid
dense region of DNA in a bacterium, not bounded by membrane
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chromatin
complex of DNA and protein,fits into the nucleus through an elaborate, multilevel system of packing
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Chromosome Packing
DNA -> Histones(5) -> nucleosome (1st 4 histones) -> 30-nmfiber (5th hsitone H1) -> Looped domains -> Metaphase
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heterochromatin
- exist in a highly condensed state similar to that seen in a metaphase chromosome
- largely inaccessible to the machinery in the cell responsible for transcribing the genetic information coded in the DNA,
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euchromatin
distinguish it from the lesscompacted, more dispersed
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