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KMID : 0364819750130030109
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Korean Journal of Microbiology
1975 Volume.13 No. 3 p.109 ~ p.115
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Electron Microscopy of Cell Walls of Saccharomces cervisiae and Mycobacterium phlei in the process of DNA extraction
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李吉洙/LEE, Kil Soo
趙世勳/金銀壽/柳駿/CHO, She Hoon/KIM, Woon Soo/LEW, Joon
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Abstract
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DNA´s were extracted from Saccharomyces cervisiae and Mycobacterium phlei and the
damaging cell walls of these microorganisms were examined under an electron
microscope in the extraction process in which a number of physico-chemical treatments
of cells was involved.
While the DNA was easily exteacted from S. cerevisiae using conventional methods of
isolation, the DNA of M.phlei was extremely difficult to isolate and yielded very little
DNA, applying various methods of isolation published earlier.
When the cell walls of S. cerevisiae were examined with the electron microscope, they
were not yet damaged even after the cells were treated with sodium lauryl sulfate(SLS)
and ethylene diamine tetraacetic acid(EDTA), but they were completely destroyed by the
treatment of sodium perchlorate followed by the addition of chloroform and a vigorous
agitation. Oozing cytoplasm through the broken cell walls was also observed.
In the extraction of DNA from M.phlei, the pronase was not effective at the aerobic
environment of the sample.
When phenol was applied at the last step of DNA isolation, an extreme damage of cell
walls was observed but the cytoplasm was shrunk into an electron-dense mass tyielding
little DNA into the solution.
Unlike the cells of S.cerevisiae, M,phlei cells showed a tendency of aggregation, thus the
destruction of cell walls by sodium hydroxide was seen only on the walls of peripheral
cells in the aggregated mass, leaving the walls of the inner cells undamaged.
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