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Section 2Molecular and Morphological Structure of Chromosomal DNA
Experiments proving that Chromosomes Posses the DNA:•Chemical analysis (Ris & Marasky)•Feulgen’s reaction•Ultraviolet absorption•Enzymatic digestion Lambproush chromosomes•Autoradiography
Section 2Molecular and Morphological Structure of Chromosomal DNA
1- Enzymatic Digestion of Lampbrush Chromosomes:
A Lampbrush chromosome is the largest chromosome known and is found in the amphibian oocytes (immature eggs). Lampbrush chromosomes occur during the diplotene stage of meiosis I. They are meiotic bivalents, each consisting of 2 sister chromatids. Lampbrush chromosomes are clearly visible even in the light microscope, where they are seen to be organized into a series of large chromatin loops emanating from a linear chromosomal axis.
Rnase +proteinase Matrix disappeared DnaseChromosome
lyses
Section 2Molecular and Morphological Structure of Chromosomal DNA
2- Feulgen’s Reactiona specific staining procedure for DNA: mild acid hydrolysis makes the aldehyde group of deoxyribose available to react with schiffs reagent to give a purple colour.
Warm and diluted Acid
DNA hydrolysis
Schiff’s reagent+
Nucleic Acid Source(CHO- Ribose)
magenta or purple color
develops
Section 2Molecular and Morphological Structure of Chromosomal DNA
3- Ultraviolet AbsorptionDNA can absorb UV waves in 2600 Å (260n.m) wave length due to the cyclic structure of the nitrogen bases.
High radiatio
n
Low
rad
iation
Chemical Analysis (Ris and Marasky)
90 %
The main compound
10 %
The secondary compound
45 % DNA
55 % Histone proteins
14 % RNA
2.6 % DNA
80 % Non Histone Proteins
Animal
( Cow)
Thyroid
glands
Autoradiography
Genome Organization• Eukaryotes Vs. Prokaryotes• Genome• Haploid• Diploid
http://www.youtube.com/watch?v=ke7PDxrKbj8
Eukaryotic Chromosome
http://www.youtube.com/watch?v=ke7PDxrKbj8
DNA Packaging Why package DNA?
Levels of Packing:
1-Nucleosome
2-30 nm fiber/Solenoid
3-Loops
4-Rossettes
5-Chromosome
• ChromosomesNucleosome (Level 1)A nucleosome core particle contains: – 146 base pairs of supercoiled DNA– Around a core of eight histone molecules
Histone coreTwo copies of H2A, H2B, H3 and H4– Octamer
H1 resides outside of core– Linker histone– Binds to linker DNA– Connecting one nucleosome core particle
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