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KMID : 1101220030350020170
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2003 Volume.35 No. 2 p.170 ~ p.173
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Detection of Nucleic Acid Strand in the Electron Microscope by Modified Shadowing Procedure
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Joo Kyeng-Woong
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Abstract
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Rabbit hemorrhagic disease(RHD) which was first recognized in China in 1984. Although Rabbit hemorrhagic disease virus was initially characterized as a picornavirus or a parvovirus, it is now proven to be calicivirus. Kleinschmidt and Zahn developed a method which allows the visualization of DNA molecules as smooth, flexible filaments. In this protein monolayer technique the nucleic acid molecules bind to a protein film which results from surface denaturation of a cytochrome C at an air-water interface. The surface film is then picked up and the nucleic acid molecules are examined in the electron microscope after uranyl acetate staining and heavy metal shadowing. This study is purposed to detection of RHD viral RNA structure by modified shadowing technique as basic procedure of Kleinschmidt and Zahn for observed to difference as ssRNA of RHD viral nucleic acids from other ssDNA on electron microscopy. I have experimented to chemical modified procedures that spreading and diffusion to 90% formamide and 0.2N NaOH denaturing agents, the signal is enhanced by silver solution before electron microscopic observation. It was appeared with effect to dense single strands of RHD viral nucleic acids.
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KEYWORD
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Electron microscope, Nucleic acid strand, RHD, Shadowing technique
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