• Korean Journal of Veterinary Research
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• v.36 no.4
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• pp.859-871
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• 1996

This study was conducted to elucidate the distribution of Aujeszky's disease viral nucleic acids and antigens in the central nervous system (CNS) of piglets. The first Korean isolate of Aujeszky's disease virus(ADV) that isolated from naturally infected piglets in Yang San, was inoculated into 32 day old piglets with $10^{5.9}TCID_{50}/ml$ through intranasal or intramuscular route. These piglets were sacrificed at every 24hrs for 8 days. The immunohistochemistry (IHC) was conducted to detect the viral antigens in paraffin-embedded tissue sections using avidin-biotin-peroxidase complex (ABC) method. The viral nucleic acids were detected by in situ hybridization (ISH) using ADV specific DNA probe labeled with digoxigenin. The ADV antigens were detected in reticuloendothelial cells of spleen, lymph nodes and tonsil, alveolar walls, leptomeningeal vascular walls, inflammatory foci of each organ, and nerve cells. The viral nucleic acids were detected in the spinal trigeminal nucleus and its tracts of the pons and medulla oblongata by the ISH technique. The pathways of AD viruses in CNS were determined by IHC and ISH. In the intranasally inoculated group, the viruses in nasal mucosa moved to medulla oblongata and pons through the trigeminal nerve. In case of intramuscullarly inoculated group, viruses moved to brain via lymphoid organs or spinal nerves from sciatic nerves.

• Journal of Microbiology and Biotechnology
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• v.10 no.5
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• pp.700-706
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• 2000

Iron- and zinc-containing superoxide dismutase (FeZnSOD) and nickel-containing superoxide dismutase (NiSOD) are cytoplamic enzymes in Streptomyces griseus. The sodF gene coding for FeZnSOD was cloned from genomic Southern hybridization analysis with a 0.5-kb DNA probe, which was PCR-amplified with facing primers corresponding to the N-terminal amino acid of the purified FeZnSOD of S. griseus and a C-terminal region which is conserved among bacterial FeSODs and MnSODs. The sodF open reading frame (ORF) was comprised of 213 amino acid (22,430 Da), and the deduced sequence of the protein was highly homologous (86% identity) to that of FeZnSOD of Streptomyces coelicolor. The FeZnSOD expression of exponentially growing S. griseus cell was approximately doubled as the cell growth reached the early stationary phase. The growth-associated expression of FeZnSOD was mainly controlled at the transcriptional level, and the regulation was exerted through the 110 bp regulatory DNA upstream from the ATG initiation codon of the sodF gene.

• The Korean Journal of Mycology
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• v.26 no.1 s.84
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• pp.103-107
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• 1998

The trp2 gene encoding trifunctional enzyme in Coprinus cinereus was investigated the expression in the heterothallic mushroom species. To identify the homology of trp2 gene to Schizophyllum commune and Pleurotus ostreatus, southern hybridization was performed with plasmid pHIONA8 containing C. cinereus trp2 gene as a probe, which resulted in a strong signal indicating an homologous sequence to the chromosomal DNA of S. commune. About 50 transformants per dish was appeared in the complementation test by pHIONA8 using S. commune tryptophan auxotroph as host. In the mating test between transformants and other mating type alleles, the fruiting body of S. commune was formed at $30^{\circ}C$ in $2{\sim}3$ weeks.

• Journal of fish pathology
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• v.8 no.1
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• pp.37-46
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• 1995

The amino acid analysis of polyhedrin protein and nucleotide sequence of polyhedrin gene in H. cunea nuclear polyhedrosis virus (HcNPV) genome have been studied. Polyhedrin had three polypeptide bands in SDS - polyactylamide gel electrophoresis. The major polypeptide had a molecular weight of 25 kd. The polyhedrin was composed of 17 different amino acids. HcNPV DNA was digested with EcoRI restriction enzyme and hybridized with ($\alpha^{32}P$) -labelled AcNPV polyhedrin gene cDNA. The polyhedrin gene was located on the fragment of EcoRI-H. The EcoRI - H fragment containing polyhedrin gene was cloned into the EcoRI site of pUC8 vector which was confirmed with southern blotting, and the recombinant plasmid containg polyhedrin gene was designated as hPE-H. The promoter region of polyhedrin genomic DNA was sequenced. The sequences identified as the TATA box was found at the 5' flanking region of the polyhedrin genomic DNA approximately -79 bp upstream from the transcriptional start site. But CAAT-like box was not shown near the TATA-like box in the polyhedrin gene. Four tandem repeats with the sequence 5' -CTAATAT-3' and 5'-TAAATAA-3' were found between -141 and -108 or -83 upstream and -52 bp downstream from the translation start site. About -141 bp region upstream from the translational start site was highly AT (78%) rich. The coding region for the polyhedrin starts and ends with ATG and TAA, respectively.

• Development and Reproduction
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• v.17 no.3
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• pp.215-220
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• 2013

Recently, the RNA/DNA-binding protein FUS, Fused in sarcoma, was shown to play a role in growth, differentiation, and morphogenesis in vertebrates. Because little is known about Fus, we investigated its expression pattern in murine tooth development. In situ hybridization of mouse mandibles at specific developmental stages was performed with a DIG-labeled RNA probe. During early tooth development, Fus was detected in the dental epithelium and dental mesenchyme at 11 days postcoitum (dpc) and 12 dpc. From 14 dpc, Fus was strongly expressed in the dental papilla and the cervical loop of the dental epithelium. At postnatal day 4 (PN4), Fus expression was observed in the odontoblasts, ameloblasts, the proliferation zone of the pulp, and the cervical loop. At PN14, the expression pattern of Fus was found to be maintained in the odontoblasts and the proliferation zone of the pulp. Furthermore, Fus expression was especially strong in the Hertwig's epithelial root sheath (HERS). Therefore, this study suggests that Fus may play a role in the HERS during root development.

• Microbiology and Biotechnology Letters
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• v.18 no.3
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• pp.305-308
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• 1990

In order to clone the gene coding for 3-isopropylmalate dehydrogenase of Muyveromyces fragilis, a shuttle plasmid vector pHNll4 was used. It can serve as a cloning vector in Saccharomyces cerevisiae DBY746 for other Sau3AI-cleaved DNA segment of Kluyveromyces fragilis. Two cloned fragments which complement the leu2 mutation of Saccharomyces cerevisiae and E, coli were obtained. Their length was 4.4 kb an 3.5 kb, and their orientation was opposite each other. From the fact that the two recombinant plasmids were expressed in Saccharomyces cerevisiae and E, coli, probably the two inserts had the promoter of Ktuyveromyces fi-agilis and that of Kluyveromyces fiagilis was efficiently assosiated with RNA polymerase of Saccharomyces cerevisiae and E. coli. According to the result of Southern hybridization, we thought that the cloned fragment has low homology with 3-isopropylmalate dehydrogenase coding region of E. coli and Saccharomyces cerevisiae.

• Applied Biological Chemistry
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• v.40 no.2
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• pp.101-106
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• 1997

A screening was performed to isolate the cellulose-producing microorganisms from vinegar in Korea. The isolated strain was identified as Acetobacter sp. with respect to physiological and biochemical characteristics and designated as Acetobacter CBI-2. Cellulose production of Acetobacter CBI-2 was equal with the well known cellulose-producing bacteria, A. xylinum. The result of separation on thin layer chromatography(TLC) was consistent with the degradation product of native cellulose. The presence of genes required for the cellulose biosynthesis in Acetobacter CBI-2 was confirmed by Southern hybridization.

• Journal of Korean Society of Forest Science
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• v.83 no.1
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• pp.20-24
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• 1994

In woody species with a long life span, the studies on inheritance of any trait may be very time consuming and laborious. Chloroplast DNA(cpDNA) has been a valuable tool in such studies since it has several unique features such as limited genome size and cytoplasmic inheritance. In the present study, cpDNAs from five different species of Populus(P. alba, P. glandulosa, P. alba${\times}$P. glandulosa, P. davidiana, and P. nigra), and Nicotiana tabacum were compared with regard to restriction fragment length polymophism. The results showed that cpDNAs among the species were very conserved, although some polymorphisms were observed when the DNAs were digested with restriction enzyme EcoRI or KphI. The other enzymes (Bgl II, and PstI) tested produced identical restriction fragmentation pattern among the species. However, cpDNAs from all the five Populus species showed different restriction fragmentation pattern from that of tobacco with the four restriction enzymes tested. Southern hybridization with tobacco rbcL gene fragment as a probe also produced identical pattern among Populus species. The results indicate that cpDNAs in the genus are very well conserved during evolution.

• Korean Journal of Bronchoesophagology
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• v.5 no.1
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• pp.49-54
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• 1999

The etiology of inverted papilloma(IP) remains unknown, but several studies have reported that Human Papillomavirus(HPV) may play a role in the pathogensis of sinonasal inverted papilloma(IP). And recent reports demonstrate the possible etiologic role of Epstein-Barr virus (EBV) in sinonasal IP. The aim of this study is to detect HPV and EBV in sinonasal IP, to examine the relationship between HPV subtype and sinonasal IP, to investigate the relation between HPV and EBV. We reviewed 30 cases of sinonasal IP(simple IP 19 cases, IP with dysplasia 8 cases, IP with squamous cell carcinoma 3 cases). Paraffin embedded archival tissue was used in this study. Detection of HPV, EBV were examined by in situ hybridization(ISH) using HPV type 6/11, 16/18, 31/33/35 DNA probe and EBER probe. The HPV was detected in 6(20%) out of 30 cases. The HPV 6/11 was dectected in 4 out of 19 cases of simple IP, HPV 16/18 in 1, HPV 31/33/35 in 1 out of 8 cases of IP with dysplasia respectively. The EBV was not detected in 30 cases. HPV may play a role in the pathogensis of sinonasal inverted papilloma. But EBV is not a etiopathologic factor to be considered in the development of sinonasal IP.

• Biomedical Science Letters
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• v.6 no.1
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• pp.65-72
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• 2000

In order to improve the early diagnosis of Johne's disease in ruminants, duplex polymerase chain reaction system for the detection of the etiologic agent of M. paratuberculosis and for the differentiation of other mycobacterial animal pathogens, such as M. bovis and M. avium, was applied. Genomic DNAs were purified from peripheral blood monocytes or milk macrophages and were used as templates in the duplex PCR. Detection of Mycobacterium spp. in the specimen was carried out by PCR using primer set specific to the mycobacterial 16S rDNA. And then, mycobacterial DNA-positive specimens were further differentiated with duplex PCR system which was composed of primer sets specific to 16S rDNA of M. avium complex and Is900 gene of M. paratuberculosis. The results were re-confirmed by Southern blot hybridization with oligonucleotide specific to the internal sequence of IS900 PCR amplicons. The applicability of this duplex PCR system was evaluated with DNAs extracted from clinical specimens of peripheral blood monocytes and milk macrophages. In summary, the duplex PCR amplification system described in this experiment is promising molecular technique for the early diagnosis of Johne's disease in ruminants.