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Lee, Gunsup,Cho, SeungChan,Hoang, Phuong Mai,Kim, Dongjun,Lee, Yongjun,Kil, Eui-Joon,Byun, Sung-June,Lee, Taek-Kyun,Kim, Dae-Hyun,Kim, Sunghan,Lee, Sukchan Korean Society for Molecular and Cellular Biology 2015 Molecules and cells Vol.38 No.9
3D8 single chain variable fragment (scFv) is a recombinant monoclonal antibody with nuclease activity that was originally isolated from autoimmune-prone MRL mice. In a previous study, we analyzed the nuclease activity of 3D8 scFv and determined that a HeLa cell line expressing 3D8 scFv conferred resistance to herpes simplex virus type 1 (HSV-1) and pseudorabies virus (PRV). In this study, we demonstrate that 3D8 scFv could be delivered to target tissues and cells where it exerted a therapeutic effect against PRV. PRV was inoculated via intramuscular injection, and 3D8 scFv was injected intraperitoneally. The observed therapeutic effect of 3D8 scFv against PRV was also supported by results from quantitative reverse transcription polymerase chain reaction, southern hybridization, and immunohistochemical assays. Intraperitoneal injection of 5 and $10{\mu}g$ 3D8 scFv resulted in no detectable toxicity. The survival rate in C57BL/6 mice was 9% after intramuscular injection of 10 $LD_{50}$ PRV. In contrast, the 3D8 scFv-injected C57BL/6 mice showed survival rates of 57% ($5{\mu}g$) and 47% ($10{\mu}g$). The results indicate that 3D8 scFv could be utilized as an effective antiviral agent in several animal models.
TYLCV transmission by whiteflies, weeds and seeds
Kyeong-Yeoll Lee,Jae-Kyoung Shim,Sukchan Lee,Eui-Joon Kil,Gwan-Seok Lee,Hong-Soo Choi,Chang-Seok Kim,Jung-Hwan Lee,Ji-Kwang Kim,Kyung-Bon Koo 한국응용곤충학회 2017 한국응용곤충학회 학술대회논문집 Vol.2017 No.04
Tomato yellow leaf curl virus (TYLCV) causes a highly serious disease in tomatoes in many countries. The most important thing in management is the prevention of virus transmission. TYLCV had been known to be only transmitted by a vector, the sweetpotato whitelfy, Bemisia tabaci. However, we identified two other important routes of TYLCV transmission into tomatoes. It is seed-transmissible. Many seeds available in domestic and global markets were infected by TYLCV. It is also infective into various weeds and other horticultural crops and transmit virus through vector insects. Therefore, we need a new strategy for the virus and vector management.
Temperature effects on plant virus transmission of vector insect Bemisia tabaci
Kyeong-Yeoll Lee,Gwan-Seok Lee,Chang-Seok Kim,Sukchan Lee 한국응용곤충학회 2013 한국응용곤충학회 학술대회논문집 Vol.2013 No.10
Acquisition of plant viruses has various effects on physiological mechanisms in vector insects. Bemisia tabaci is the only known vector of Tomato yellow leaf curl virus (TYLCV), which is a serious virus affecting tomato cultivars. In this study, the lifespan of Q1 biotype was compared between non-viruliferous (NV) and TYLCV-viruliferous (V) whiteflies. Total lifespan from egg to adult death of NV whiteflies was 62.54 days but 10.64 days shorter in V whiteflies. We investigated the temperature susceptibility of B. tabaciby comparing mortalities as well as heat shock protein (hsp) mRNA levels between NV and V whiteflies. For this, NV and V whiteflies were exposed for either 1 or 3 h at 4, 25, and 35 °C. The mortality of V whiteflies was higher than NV ones following exposure at either 4 or 35 °C, but there was no significant difference at 25 °C. Analysis of the expression level of heat shock protein (hsp) genes using quantitative real-time PCR showed that both cold and heat shock treatments stimulated higher expression of hsps (hsp40, hsp70, and hsp90) at various rates in V whiteflies than NV ones, but there was no difference at 25 °C. All together, our results show that TYLCV acquisition accelerated the developmental rate and increased susceptibility to thermal stress in B. tabaci. Therefore, this modification may result in reduced vector longevity due to increased metabolic energy utilization. Our results provide insights into the complex interaction between vector fitness and thermal stress in relation to the acquisition and transmission of plant viruses.
Kyeong-Yeoll Lee,Woo-Geun Song,Sukchan Lee,Eunseok Choi 한국응용곤충학회 2008 한국응용곤충학회 학술대회논문집 Vol.2008 No.10
The sweet potato whitefly Bemisia tabasi is one of the most important pests of various horticultural crops. In addition, B. tabaci is a vector of many plant-pathogenic viruses and cause a serious secondary damage to crop plants. Association of plant-pathogenic virus with vector insects is known to be effective on the transmission capacity, fecundity, longevity of vectors including whiteflies. However, the interactive mechanisms between virus and vector insects are still poorly understood. Recently, a serious damage caused by virus disease together with B. tabasi emergence was identified at tomato glasshouse in Tongyoung. We detected the signals of Tomato Yellow Leaf Curl Virus (TYLCV) in tomato leaves and vector whiteflies using PCR amplification and confirmed its presence by those sequence comparison. To determine the effects of TYLCV acquisition on physiological status of vector whiteflies, transcript levels of genes that associated with metamorphosis, metabolism, stress and immune processes were compared between TYLCVinfected whiteflies and non-infected ones. Generally, the transcript levels of virus-infected whiteflies were lower than those of non-infected ones. In addition, the associations of endosymbiont levels within whiteflies were discussed in aspect of the acquisition and transmission of TYLCV.
Gwan-Seok Lee,Seol-Mae Lee,Hae-Ryun Kwak,Chang-Seok Kim,Hong-Soo Choi,Sukchan Lee,Kyeong-Yeoll Lee 한국응용곤충학회 2012 한국응용곤충학회 학술대회논문집 Vol.2012 No.10
The sweet potato whitefly, Bemisia tabaci (Gennadius, 1889) (Hemiptera: Aleyrodidae), is one of the most important agricultural pests by direct feeding and begomovirus transmission in the world. B. tabaci species complex consisting of about 30 species worldwidely has been mainly distinguished by molecular methods because of difficulties detecting morphological differences. In Korea, distribution of two exotic groups, the Mediterranean (Q biotype) and the Middle East-Asia Minor 1 (B biotype), were confirmed by comprehensive molecular methods in 2012. Of them, the Q biotype as a vector of Tomato yellow leaf curl virus (TYLCV) has caused significant crop yield losses. Tobacco leaf curl virus (TbLCV), one of four begomoviruses in Korea, was first reported in 2011. It damaged, although not frequent, to greenhouse tomatoes at Iksan, Gimje, Sunchang, and Jeju. The vector transmitting TbLCV, however, is not known yet. In this study, we sampled an unknown B. tabaci populations on leaves of the Japanese honeysuckle (Lonicera japonica) at Iksan in outdoor conditions. PCR analysis revealed that some populations were infected with TbLCV. We identified an unknown indigenous genetic group named as JpL and clarified its phylogenetic relationships based on nucleotide sequences of mitochondrial cytochrome oxidase I gene.
Morphology and Anatomy of Mature Embryos and Seedlings in Parasitic Angiosperm Cuscuta japonica
Lee, Kyu Bae,Park, Jong-Bum,Lee, Sukchan 한국식물학회 2000 Journal of Plant Biology Vol.43 No.1
Morphological and anatomical features of mature embryos and seedlings were observed at different growth stages in the parasitic angiosperm Cuscuta japonica Choisy. The spirally coiled embryos from scarified seeds had no cotyledons but possessed blunt radicles. Seeds germinated at 30℃ in the dark. Although most embryo cells incubated for 16 h did not have starch grains, the shoot cells of three-day-old seedlings possessed numerous starch grains. After these seedlings were transferred to a lightened growth chamber, all the shoot apical regions of seedlings grown for 6,8, and 10days became greenish and hooked. Most of the shoot cells, including the green apical parts, contained abundant starch grains. The hooks opened only when one seedling made contact with another seedling. This suggested that the green and hooked shoot apical regions played an important role in searching for and twining about their host plants. In some two-day-old seedlings, the massive roots were circular or semi-circular. This enabled the shoot axes to stand erect on some substratum. It would assist the shoots in making contact with the host plant. In eight-day-old seedlings, the green apical regions also were hooked and the roots were considerably degraded.
Expression of influenza virus neuraminidase on tobacco for edible vaccine development
Lee, Kyung-Real,Lee, Sukchan 성균관대학교 생명공학연구소 2000 生命工學硏究 Vol.6 No.1
Oral vaccine is the vaccine which can immunize against antigen proteins by means of putting them by orally. Especially, it has recently been studying about 'edible vaccine' which able to give immunity against antigen proteins by just eating plants produced them, no purification. The merits of this are economical, saving manpowers and more efficient preventing diseases. We made plans for expression of neuraminidase (NA), antigen protein o finfluenza virus, on to tobacco plants using agrobacterium transformation to examine production of edible vaccine. Putative transgenic tobaccos were regenerated and 1.4kb of NA-RNA were found from the total RNA of transplants using RT-PCR. Further experiments are attempted to raise stability of expression targeting into ER or chloroplast and to confime edible vaccine production by bioassay using plant diets.
Isolation and Expression of Red Algae Phycoerythrin on Yeast
Lee, Jungwoo,Lee, Sukchan 성균관대학교 생명공학연구소 2000 生命工學硏究 Vol.6 No.1
It is increasing necessity of microorganisms for mass production of natural pigments for food application because of both increase of natural pigments demands and liking of natural foods. The phycoerythrins contents of the phycobilisomes of the marine algae of pophyra sp. This work reports the characterization of gene encoding PE in the red alga porphyra sp. The nucleotide sequence of approximately 1.1kb which contain two open reading frames of 565 and 496 nucleotides. These genes were named rpeβ and rpeα. The arrangement of the coding sequences parallels that of the Rodophyta PE genes, with rpeβ and rpeα located 5' of rpeα. The two genes are separated by an intergenic region of 76 nucleotides, and both are transcribed from the same DNA strand. cDNA sequences coding RPE was a member of a homelogous family of peptides, the phycoerythrin. The RT-PCR product of RPE gene was cloned and expressed in yeast.