| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1990: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1989: ¥1,500,000 (Direct Cost: ¥1,500,000)
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| Research Abstract |
Immunocytochemical localization of structural and non-structural proteins of tobacco mosaic virus (TMV) in tobacco protoplasts and brome mosaic virus (BMV) in barley cells were investigated by protein A-gold technique.
TMV : When utilizing the anti-coat protein antiserum, gold labels were first found in small areas in the cytoplasm 4 hr after inoculation and followed immediately by occurrence of virus particles. Subsequently, virus particles formed crystalline arrays and during the period of active virus multiplication, labels were confined only on this virus crystalline aggregates. In later stages of infection, lighter labels also were found throughout the cytoplasm where no virus particles were evident. Nucleus, plastid and mitochondria were free of labels throughout infection. Labels with antiー130K protein antiserum were first detected on virusーinduced small inclusions in the cytoplasm 4 hr after inoculation. Several inclusions in a protoplast occurred, and they are often in the vici
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nity of virus aggregates. The rapid increasing in the size of the inclusion were take placed during later stages of infectin, in which virus replication are slowdown, rather than during vigorous production of virus. Labels were always associated with this inclusions. The labels with antiー180K protein antiserum were also dispersed over this inclusions. When sections were treated with antiー30K protein antiserum, specific gold labels were first observed at several small regions in the cytoplasm at 4-6 hr after inoculation. Thereafter, the number of labeling areas increased along the time course. When protoplasts were cultured in the presence of actinomycin D (30mug/ml). Large areas in the cytoplasm were heavily labeled. These labeling areas were slightly less electronーdense than surrounding cytoplasm and fulled of granular structures.
BMV : When the coat protein antiserum were used, much gold labels were observed through out the cytoplasm of barley cells. In particular, heavy labels were found on the region of high concentration of virus particles in the cytoplasm. Specific but less dense labeling was observed over some nuclei infected cells. However, specific labeling was not present inside chloroplast, mitochondria and microbodies. Gold labels of 3a protein were found over the region where very high concentrations of vurus particles occur in the cytoplasm. Less
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