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Fluorometrical detection of thymine base differences in complementary strands of satellite DNA in human metaphase chromosomes

โœ Scribed by My. A. Kim


Publisher
Springer
Year
1975
Tongue
English
Weight
716 KB
Volume
28
Category
Article
ISSN
0340-6717

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โœฆ Synopsis


Using the fluorochrome "Hoechst 33258", intensity of fluorescence was found to differ distinctly between the sister chromatids in the paracentric regions of chromosomes 1, 16, and 19, after one round of replication in medium containing BUdR. Thus the effect of fluorescence asymmetry is not limited to the part of the Y chromosomes that fluoresces intensely with quinacrine; it can also be determined in the weakly Q-fluorescent pericentric regions of chromosomes, which are known to be the sites where highly reiterated sequences of satellite DNA are located. However, an exception is the paracentric region of chromosome 9 which does not show the effect of lateral asymmetry. The difference of fluorescence intensity in the heterochromatic regions of the sister chromatids of human chromosome 1 is measured by densitometric tracement along the long axes of chromosomes; this is obtained from two individuals with an "uncoiler" heterchomatic block (type III) having a relative intensity of 1:1.93 in an average of the total measured blocks. This corresponds to the uneven distribution of thymine base of 22.8 and 43.2 in the two strands of the DNA double hexlix. A chromatid exchange rate of 9 in 100 metaphases per cell cycle was found within the uncoiler region of chromosome 1.


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Human satellite DNAs I, II and IV were transcribed to yield radioactive complementary RNAs (cRNAs). These cRNAs were hybridised to metaphase chromosomes of man, chimpanzee (Pan troglodytes), gorilla (Gorilla gorilla) and orang utan (Pongo pygmaeus). The results of this in situ hybridisation were ana