• Animal cells and systems
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• v.2 no.4
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• pp.529-538
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• 1998

In order to analyze the intracellu1ar localization of specific RNA components of ribonucleoproteins (RNP) in Xenopus oocytes, a modified protocol of whole-mount in situ Hybridization is presented in this paper, Mitochondria specific 12S rRNA probe was used to detect the amplification and distribution of mitochondria in various stages of the oocyte life cycle, and the results were found to be consistent with previously known distribution of mitochondria. The results with other specific probes (U1 and U3 small nuclear RNAs, and 5S RNA) also indicate that this procedure is generally effective in localizing RNAs in RNP complexes even inside organelles. In addition, the RNA component of RNase MRP, the RNP with endoribo-nuclease activity, localize to the nucleus in various stages of the oocyte life cycle. Some of MRP RNA, however, were found to be localized to the special population of mitochondria near the nucleus, especially in the active stage of mitochondrial amplification. It suggests dual localization of RNase MRP in the nucleus and mitochondria, which is consistent with the proposed roles of RNase MRP in mitochondrial DNA replication and in rRNA processing in the nucleolus.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.45 no.2
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• pp.143-149
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• 2000

From the soils of soybean fields in Cotton Branch Station (CBS) and Pine Tree Station (PTS), Arkansas, USA, various single spore isloates of sudden death syndrome (SDS) pathogen were obtained on modified Nash ＆ Snyder's medium (MNSM) with dilution plating technique and transferred to potato dextrose agar (PDA) medium to identify the cultural colony shape. The colony shapes of these isolates resembled F. solani isolate 171 which was white and chalky shaped on MNSM and most of them had unique form of morphology which produced white margin and blue center colony on PDA. Although, some of these isolates had more dark blue or showed slightly different color, all isolates that were selected randomly for green-house inoculation assay produced typical foliar symptoms on leaves of soybean, Hartz 6686. To determine the genetic differences among the isolates, mitochondrial DNA restriction fragment length polymorphism (RFLP) was conducted with fourty isolates from both fields, using mtDNA probes, 2U18 and 4U40, derived from Colletotrichum orbiculare. We obtained distinctive RFLPs in each treatment of restriction enzyme, EcoRI and HaeⅢ. Isolates, 11-2-5 and 14-3-1-1, from CBS and isolates, 104-3-1-2 and 701-1-5-1, from PTS showed different band patterns from 171 in both or in either treatment of restriction enzymes. Even if some of these isolates showed heterogeneous, they were more closer to 171 than PN603. And, also, rest of the thirty-six isolates had exactly same polymorphisms as 171 in each treatment of restriction enzyme. Although, some of the isolates showed the different morphological shape on PDA and slightly different band patterns on RFLPs, all of the isolates selected on MNSM due to their distinctive colony shape from other fungi produced the typical foliar symptoms on soybean leaves in greenhouse inoculation assay. It might be suggested that these isolates were not genetically different from check isolate 171 and they were unique strain of F. solani.

• Biomedical Science Letters
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• v.2 no.2
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• pp.257-265
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• 1996

Simple stain and electron microscopic in situ hybridization is studied and applied for the identification of rabbit haemorrhagic disease viral RNA in a unicrylated preparation of the liver after innoculation of rabbit haemorrhagic disease virus. Hybridization for detection of viral RNA in unicryl embedded tissues using complementary 84 bases oligonucleotide probe labelled by biotin CE-phosphoramidite compared with 4717∼4800 sequences of rabbit haemorrhagic disease virus, modified hybridization protocol and antibiotin antibody-l0nm gold as signal marker. The best results were obtained in 0.02% glutaraldehyde, Unicryl resin cell block, biotinylated oligonucleotide probes, antibiotin-l0nm gold. In this report, RHD viral RNA was distributed widely within the mitochondria and nucleus of liver cell by electron microscopic in situ hybridization. In situ hybridization has become a standard method for localizing DNA or RNA sequences in tissue or celt preparation. In situ hybridization is detected the virus genome in the cells and tissue as specifically compared with others nucleic acid hybridization method.