DNA as genetic material 1
- Created by: mayasheryn
- Created on: 02-04-18 10:26
genes
- carried on chromosomes
- linked closely- unless they are distant on loci
transformation 1
Griffith (1929) s.pneumoniae is lethal in mice
2 strands= smooth and rough
smooth -> wild type and lethal
rough -> mutant and non lethal
transformation 2
smooth bacteria killed by boiling and injecting into the mice = mice survived
-> BUT inject R and dead S cells = mice die as R turned into S = transformation
what caused it to transform?
DNA tranforming principle
Avery experiment
detergent to lyse heated S
- removed proteins and combined with R= mice die
- removed RNA and combined with R= mice die
- removed DNA and combined with R= mice live
Hershey chase experiment
- DNA = high P
- protein= high S
T2 put on bacteria when radioactive 32P= therefore DNA molecule of inheritance
pyridamines
T
C
U
purines
A
G
RNA
D-ribose
unstable to due to extra -OH
DNA
2-deoxyribose
nucleosides
ribose or deoxyribose linked to purine or pyridamine
- A + ribose= adenosine
- U + ribose= uridine
- G + 2-deoxyribose = deoxyguanosine
- C + deoxyribose = deoxycitidine
nucleotides
- phosphoric acid ester of a nuceloside
- ribsonucleosides have 3 free -OH groups of sugar ring- 3-ribonuceoside monophosphates are possible
problems with DNA
pyridamine has to eqaul purine
nucleic acid structure
- 5' and 3' structure- designed to detect where C atoms are
secondary structure
watson and crick (1953) using x-ray
B-DNA- H bonds between bases -> antiparrallel in sugar ring as distinct strands from bases
- proteins make it easy to fix by detector
chemical analysis of DNA
- higher the G-C= higher density
- typical mRNA is about 1.5 Kb long
- dyes and antibiotics have stronger binding specificities for G-C or A-T
DNa denaturation and hyperchromic effect
- H bonds effected by temp or pH
- dsDNA collapses into **DNA and is called helix-coil tranisiton
- the temp at which ds-> ** is called the Tm
- hyperchromic effect causes materials to decrease in their ability to absorb light
factors affecting DNA denaturation
- nature of DNA: homogeneous DNA melts at a lower temo range then heterogeneous
- helix-coil transition changes denisty of DNA= ssDNA has a higher density of G-C than dsDNA
- DNA denaturation and Cot value analysis
- when ssDNA is returned to physiological conditions is may reassociate to for a double helix
- however renaturation rates and completion mare affected by several parameters
DNA replication
- semi conservative
- meselson and stahl
- E.Coli in medium with heavy nitorgen 15N not light N
- then shifted the cultures into normal medium and isolated samples after 1-2 division cycles
- prolonged centrifugation of CsCl a density gradient with the highest ion conc at the bottom of the tube
- dneisty DNA is typically a funciton of GC/AT base composition but addition of 15N increases density of DNA relative to that with 14N
and intermediate density bamd was formed between heavy and light band of the E.Coli
Meiosis
E.coli do not undergo meiosis
Taylor tested this with 3H thymidine labelling in grasshopper testes followed by slide-mounting and autoradiogrpahy
harlequin chromosomes
putting genetic map onto DNA
Phage Y genome has a linear 50Kb genome which can be readily be sheared by pipetting breaking the genome approx in the middle of the molecule giving fragments that differ in their GC content and then density= separation
- KAISER introduced the wild type halves of this DNA into different bacteria culters simulataneously with infection with phage Y mutant for the same genes
- then introduced lytic replicaiton by Y and recovered phage that were wild type for the markers on each hand of the genome
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