• Nuclear Medicine and Molecular Imaging
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• v.43 no.5
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• pp.468-477
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• 2009

Purpose: Hydrodynamic-based procedure is a simple and effective gene delivery method to lead a high gene expression in liver tissue. Non-invasive imaging reporter gene system has been used widely with herpes simplex virus type 1 thymidine kinase (HSV1-tk) and its various substrates. In the present study, we investigated to image the expression of HSV1-tk gene with 5-(2-iodovinyD-2'-deoxyuridine (IVDU) in mouse liver by the hydrodynamicbased procedure. Materials and Methods: HSV1-tk or enhanced green fluorescence protein (EGFP) encoded plasmid DNA was transferred into the mouse liver by hydrodynaminc injection. At 24 h post-injection, RT-PCR, biodistribution, fluorescence imaging, nuclear imaging and digital wholebody autoradiography (DWBA) were performed to confirm transferred gene expression. Results: In RT-PCR assay using mRNA from the mouse liver, specific bands of HSV1-tk and EGFP gene were observed in HSV1-tk and EGFP expressing plasmid injected mouse, respectively. Higher uptake of radiolabeled IVDU was exhibited in liver of HSV1-tk gene transferred mouse by biodistribution study. In fluorescence imaging, the liver showed specific fluorescence signal in EGFP gene transferred mouse. Gamma-camera image and DWBA results showed that radiolabeled IVDU was accumulated in the liver of HSV1-tk gene transferred mouse. Conclusion: In this study, hydrodynamic-based procedure was effective in liver-specific gene delivery and it could be quantified with molecular imaging methods. Therefore, co-expression of HSV1-tk reporter gene and target gene by hydrodynamic-based procedure is expected to be a useful method for the evaluation of the target gene expression level with radiolabeled IVDU.

• Korean Journal of Microbiology
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• v.41 no.1
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• pp.24-37
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• 2005

This study was conducted to analyze the molecular epidemiological properties and to select the most efficient and reliable PCR method on 116 of Staphylococcus aureus (S. aureus) isolates from Korean cattle, black goat, pig, dog, chicken, mouse and also human clinical cases from hospital. The distribution patterns of SSG [species specific genes; coagulase (coa), protein A (spa), nuclease (nuc) and aroA (RsaI) gene] were analyzed by PCR method. Among the SSGs, the nuc-gene was found in all strains $(100\%)$ tested and followed by coa-gene $(87.9\%)$, spa-gene $(91.4\%)$ and aroA-gene $(26.7\%)$, in order. The genetic subtyping by RFLP method was performed on the coa [AluI] and aroA-gene [RsaI] PCR products. The mecA-gene PCR and PCR-RFLP techniques were chosen to detect and verify of MRSA strains. Only the human strains $(12.1\%)$ were detected the positive mecA-gene products (533 bp), which were divided into two specific bands [201 & 332 bp] by HhaI enzyme digestion. On coa-gene and spa-gene typing, coa-gene was typed with ten kinds of genotype and coa-3 type were determined as the most predominant genotype, while spa-gene was divided into eleven kinds of genotype and also spa-7 type were selected the most prevalent genotype based on their genetic variations. On the aroA and coa-gene subtyping by PCR-RFLP, aroA-gene products were discriminated with only seven types of genotype, while coa-gene products were further divided into an eleven genotype, respectively. In comparison of SID values of five PCR based typing methods, the coa-PCR-RFLP (SID0.894) was evaluated the most efficient and reliable tools and followed by coa-PCR (SID0.883) and aroA-PCR-RFLP (SID0.462), in order. In conclusion, we could determined that the coa-PCR-RFLP method was the most suitable genetic analysis tool for S. aureus and MRSA strains from domestic animals and humans.

• Journal of the Korean Society of Food Science and Nutrition
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• v.34 no.5
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• pp.619-625
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• 2005

This study has been carried out to examine the characteristics of anti-H. pylori antibodies of milk produced from cows immunized with antigen of Helicobacter pylori such as peculiarity of antigen antibody, agglutination of H. pylori strain, stability of antibody against acid, alkali and heat treatments. The molecular weight of anti-H. pylori antibody measured by SDS-PAGE were turned out as about 50 kDa in the heavy chain and about 24 kDa in the light chain. Twenty protein bands were visualized in H. pylori interacting with anti-H. pylori antibody which was made in dairy cow by immunization with H. pylori. The western blotting was peformed in order to examine the antigen peculiarity of anti-H. pylori, The results were all 7 antigen substances including serum, furified serum, whey and furified whey could be confirmed and the major antigen substances were 97, 66,34 kDa of molecular weight. As a result of agglutination response anti-H. pylori antibody in whey showed 1/10 agglutination value against H. pylori. In stability test about acid and alkali of antibody, there was $100\%$ activated at the range of $pH5\~pH10$. In stability test about heat, the antibody showed stable condition at $60^{\circ}C$ for 60 minutes and comparatively stable condition at $70^{\circ}C$, but reduced activation to $40\%$ after 60 minutes. It maintained $77\%$ activation at $80^{\circ}C$ for 4 minutes and comparatively stable at $100^{\circ}C$ for 1 minute.

• Research in Plant Disease
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• v.14 no.1
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• pp.15-20
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• 2008

Three isolates of Cucumber mosaic virus (CMV) were isolated from weed hosts showing typical mosaic symptoms, and some properties of the viruses were investigated. CMV isolates, designated as Is-CMV, Jd-CMV and Pla-CMV from Isodon inflexus, Jeffersonia dubia and Phryma leptostachya var. asiatica, respectively, were identified and characterized by biological reaction in several host plants, serological property, dsRNA analysis, reverse transcription-polymerase chain reaction (RT-PCR), restriction fragment-length polymorphism (RFLP). All isolates systemically infected in Nicotiana benthamiana, Cucurbita pepo cv. Black beauty and Cucumis sativus, and did not reveal any differences in these host plants between the isolates. However, remarkable difference in the symptoms was found between the CMVs in Capsicum annuum. Is-CMV induced an asymptomatic symptoms, while Jd-CMV and Pla-CMV produced severe mosaic symptoms in C. annuum plants. In dsRNA analysis, all isolates revealed four major bands with estimated molecular size of 3.4, 3.2, 2.1 and 1.0 kbp. The cDNAs of coat protein gene of the isolates were amplified by RT-PCR using a genus-specific single pair primers that designed to amplify a DNA fragment of approximately ranging from 938 to 966 bp. By restriction mapping analysis using RFLP of the RT-PCR products as well as by serological properties of gel diffusion test, the CMV isolates belong to a typical members of CMV subgroup IA. This is the first report on the occurrence of CMV in the three weed hosts.

• Journal of Sericultural and Entomological Science
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• v.24 no.2
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• pp.55-72
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• 1983

These studies were done to find out any difference, ultrastructural, physical or chemical, between the shells of diapausing and non-diapausing eggs of the silkworm, Bombyx mori L. 1. From the electron-microscopic observation, the egg shells have four distinctive layers. In addition to the four layers, the shells in the diapausing eggs has another layer with low electron density on its surface. 2. The permeability of the egg shell to hydrochloride was much lower in diapausing egg than in non-diapausing egg. Also the permeability changed in the opposite directions with the egg age: the diapausing eggs decreased while non-diapausing ones increased. 3. The permeability increased when the diapausing egg shell was treated with HCl. When they were treated with ether, however, the increase in permeability was much smaller. It seems there was an ether soluble material involved in the content of the egg shell. 4. The diapausing eggs were also much more resistant to desiccation than the non-diapausing ones. The former, when treated with HCl or chilling, became less resistant to desiccation. 5. The positive histochemical response of the egg shell to PAS-Alcian blue and protein stainings suggests presence of abundant proteins and carbohydrates in the egg shell. On the other hand, the staining response to lipid was more positive in the inner layers than in the outer layer of the shell. 6. The egg shell adhesives seems to be mucopolysaccharides produced by colleterial glands, since the oviposited eggs showed a positive responses to carbohydrate and negative to lipid-staining chemicals, but not the mature oocytes in the ovarioles. 7. There were two bands on the electrophoretic pattern of the SH proteins extracted from the egg shells both in the diapausing egg and non-diapausing one: a slow moving major component and a fast moving minor one. However, the electrophoretic mobility showed a difference in the minor components between them. It is evident that the fast moving minor one of non-diapausing egg ran a little further than that of diapausing egg. 8. In amino acids analysis, no significant differences were found in their composition between diapausing and non-diapausing egg and SH proteins contain relatively more glycine and less cystine.

• Journal of Plant Biotechnology
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• v.45 no.4
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• pp.375-381
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• 2018

The consumption of oats (Avena sativa L.) with high nutritional utility is accelerating due to the increased consumers' demand for functional foods. In Korea, naked oats are used as food, while covered oats are used for animal feed. However, it is difficult to distinguish naked oats from covered oats when the husk is removed from the grains by a special process. The present study was carried out to investigate experimental methods that would be beneficial in the segregation of different types of oats after husk removal. Grain quality-related biochemical compounds were analyzed in a bid to differentiate the oat dehulling characteristics. In addition, 61 SSR markers were examined for genetic relationship and variety identification of oats using five naked and seven covered oat varieties. Results showed that, the contents of protein, lipid, and ${\beta}-glucan$ were not significantly different among the oat varieties and this could not be used as an index for distinguishing oats husk character. However, in the fatty acid composition ratio,, naked oats had a higher ratio of stearic acid (C18:0) and oleic acid (C18:1) than covered oats, and covered oats had a higher ratio of linoleic acid (C18:2) and linoleic acid (C18:3) than naked oats. The assessment of SSR marker genotype revealed that 33 polymorphic bands among 12 oat varieties and 1 variety could be distinguished through the combination of polymorphic markers thus indicating the usability of these markers for variety identification in oats.