Purpose: Small size of recombinant scFv antibody has many advantages such as rapid blood clearances and improved targeting antibodies to tumor region. On the other hand owing to small size, number of amino group is insufficient in conjugation with chelator and fluorescence labeling. This study is to introduce poly lysine tag to the C-terminal end of scFv lym-1 sequence for fluorescence chelator conjugation. Materials and Methods: Poly lysine scFv lym-1 gene, cloned into pET-22b (+) vector, was expressed in E. coli BL21 (DE3) strain. Antibody purification was performed with Ni-NTA column and then size exclusion column chromatography. Expression and purification levels of poly lysine tagged scFv lym-1 antibody were confirmed by western blot analysis. I-124, I-125, I-131 and Tc-99m were used for radiolabeling of purified poly lysine scFv lym-1. Flow cytometry analysis of FIT( conjugated poly lysine scFv lym-1 was performed for confirmation of immunoreactivity of human Burkitt's lymphoma cells. Results: Poly lysine scFv lym-1 antibody was purified through two steps and identified as molecular weight of 48 KDa. Radiolabeling yields of I-124, I-125, I-131 and Tc-99m into poly lysine scFv lym-1 were >99%, >99%, >95% and >99%, respectively. Flow cytometry analysis of poly lysine scFv and scFv lym-1 was showed similar immunoreactivity to human Burkitt's lymphoma cells. Conclusion: Poly lysine tag was useful for the sufficient number of amino groups to scFv lym-1 antibody for chelator conjugation with minimizing loss of immunoreactivity.
Kim, Jeong-Min;Yun, Sang-Im;Song, Byung-Hak;Kim, Jin-Kyoung;Lee, Young-Min
Korean Journal of Microbiology
/
v.46
no.2
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pp.140-147
/
2010
Japanese encephalitis virus (JEV), a member of the mosquito-borne flaviviruses, causes epidemics of viral encephalitis in the Southeastern Asia. JEV is a small enveloped virus with a positive-sense RNA genome; the infectious virion consists of three structural proteins, namely capsid, membrane (M; a mature form of its prM precursor), and envelope proteins. Here, we investigated a role of the charged residues found at the N-terminus of the JEV M protein in virus production. Using an infectious JEV cDNA, we generated two mutant cDNAs, Mm1 and Mm2, by charged-to-alanine substitution for $E^9$ and $K^{15}K^{16}E^{17}$ residues of the M protein, respectively. By transfection of wild-type or each of the two mutant RNAs transcribed from the corresponding cDNAs, we found that Mm2, but not Mm1, had a ~3-log decrease in virus production, even though a comparable amount of all three structural proteins were produced in transfected cells. Interestingly, the prM protein expressed in Mm2 RNA-transfected cells was not recognized by the polyclonal antiserum raised against the N-terminal 44 amino acids of the wild type M protein, but reacted to the antiserum raised against the corresponding region of the mutant Mm2. Our results indicate that three charged residues ($K^{15}K^{16}E^{17}$) in JEV M protein play a role in virus production. Two polyclonal antisera specifically recognizing the wild-type or Mm2 version of the M protein would provide a useful reagent for the functional study of this protein in the virus life cycle.
Journal of the Korea Organic Resources Recycling Association
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v.16
no.4
/
pp.64-73
/
2008
The cDNA of CCF (coelomic cytolytic factor)-like gene (EC 3.2.1.16), a kind of glycosyl hydorlase, was isolated and cloned from the midgut of the earthworm Eisenia anderi. The size of nucleotide sequence appeared to be 1,152 bp and its predicted coding region was composed of 384 amino acid residues including the initiation methionine. The 17 residues at N-terminal end in the deduced amino acid sequence were regarded to be a signal peptide. Based on the amino acid sequence analysis, it appeared that this CCF-like protein could belong to glycosyl hydrolase family 16 (GHF16) and showed a high sequence homology of about 79~99% with CCF and CCF-like proteins from other earthworm species. The CCFs and CCF-like proteins from various earthworm species exhibited a 100% homology in the polysacchride-binding motif and glucanase motif. It has been reported that the CCFs isolated from E. fedita appeared to show a broader pattern recognition specificity than those from other earthworm species because this species resides in decaying organic matter showing very high microbial activity, implying that CCF-like protein isolated in this study from E. andrei might exhibit a broad substrate specificity that is a useful characteristic for industrial application. A phylogenetic analysis using the deduced amino acid sequences of CCF-related proteins through the BLASTX revealed that GHF16 families could be divided into three groups of metazoa, viriplantae and eubacteria subfamily. Subsequently the CCF-related proteins of metazoa subfamily could clearly be subgroup into lophotrochozoan and edysozoan type including a deuterostome origin. Further understanding of the biological properties of E. andrei CCF-like protein should be addressed to regulate the ${\beta}$-D-glucan hydrolysis and production for the industrial uses.
The parathyroid hormone related protein (PTHrP) is the most common causative peptide of humoral hypercalcemia of malignancy. In contrast, the serum level of parathyroid hormone (PTH) is low to undetectable in the majority of patients with malignancy associated hypercalcemia. Few cases exist in which the production and secretion of PTH by malignant nonparathyroid tumors have been authenticated. To our knowledge, there is very rare case in which a nonparathyroid tumor expressed simultaneously both the PTH and PTHrP. We report a case of squamous cell carcinoma of the lung with hypercalcemia which presented with simultaneous elevation of serum PTH and PTHrP. Severe hypercalcemia (serum calcium, 7.5 mEq/L) was found in a 65-year-old man who had a squamous cell carcinoma of the lung without any bony metastasis and detectable parathyroid abnormalities on isotope scintigraphy. The serum level of intact parathyroid hormone (PTH) con centration was markedly elevated as measured in two site radioimmunoreactive PTH assays (intact PTH 150 pg/mL ; normal 9~55). The serum level of a PTHrP was also increased as measured in C-terminal region specific radioimmunoassay (PTHrP 99.1 pmol/L; normal 13.8~55.3). There are no evidences of coincidental primary hyperparathyroidism in parathyroid MIBI scan and other imaging studies including neck ultrasonography and computed tomography. These results suggest that simultaneous elevation of serum PTH and PTHrP in this patient can be caused by production of both PTHrP and PTH in other nonparathyroid lesions such as squamous cell carcinoma.
The G-banding patterns of normal and of delayed spiralized chromosomes by BUdR were investigated in three established cell lines of dwarf hamsters. The results obtained were as follows: 1. The number of G-bands appeared in Chinese hamster T-233 cell line was 65. The centromeric dark band was found in No.1 chromosome and weakly stained bands were also observed in part of the centromeric regions of Nos. 2, 3, 8 and $X_2$ chromosomes. Two homologous X chromosomes were found in different banding patterns. Terminal dark bands were shown in No. 1 chromosome. No conspicuous bands appeared in No. 10 chromosome. 2. Eighty four bands appeared in Armenian hamster Y-1249 cell line. Centromeric dark bands were observed in Nos. 5 and 10 chromosomes and moderatly stained bands were also found in near the centromeric region of the long arms of Nos. 7 and 9 chromosomes. Two isomorphic X chromosomes were also distinguished by their banding patterns. 3. In Y-1313 Armenian hamster cell line, the bands were 69. No centromeric dark bands were observed in this cell line, but moderatly stained bands appeared in the centromeric area in the long arm of No. 9 chromosome. The banding patterns of these two cell lines of Armenian hamster were quite different and readily distinguished. Only No. 8 chromosome showed similar G-banding patterns. Although Nos. 5, 7 abd 8 chromosomes revealed the same number of bands in these two cell lines, the location and staining intensity were quite different. 4. Chromosomes of Nos. 1, 2, 6, $X_1$ and $X_2$ in T-233 cell line and of 1, 4, 7, 8, 9, $X_1$ and $X_2$ in both cell lines of Armenian hamster were found to be elongated due to the inhibition of mitotic spiralization by BUdR. G-banding patterns of these chromosomes were found to be identical to those of normal chromosomes in these cell lines.
Proceedings of the Korean Institute of Navigation and Port Research Conference
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2003.05a
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pp.273-280
/
2003
The purpose of this study is to identify and evaluate the competitiveness of ports in ASEAN(Association of Southeast Asian Nations), which plays a leading role in basing the hub of international logistics strategies as a countermeasure in changes of international logistics environments. This region represents most severe competition among Mega hub ports in the world in terms of container cargo throughput at the onset of the 21 st century. The research method in this study accounted for overlapping between attributes, and introduced the HFP method that can perform mathematical operations. The scope of this study was strictly confined to the ports of ASEAN. which cover the top 100 of 350 container ports that were presented in Containerization International Yearbook 2002 with reference to container throughput. The results of this study show Singapore in the number one position. Even compared with major ports in Korea (after getting comparative ratings and applying the same data and evaluation structure), the number one position still goes to Singapore and then Busan(2) and Manila(2), followed by Port Klang(4), Tanjugn Priok(5), Tanjung Perak(6), Bangkok(7), Inchon(8), Laem Chabang(9) and Penang(9). In terms of the main contributions of this study, it is the first empirical study to apply the combined attributes of detailed and representative attributes into the advanced HFP model which was enhanced by the KJ method to evaluate the port competitiveness in ASEAN. Up-to-now, none have comprehensively conducted researches with sophisticated port methodology that has discussed a variety of changes in port development and terminal transfers of major shipping lines. Moreover, through the comparative evaluation between major ports in Korea and ASEAN, the presentation of comparative competitiveness for Korea ports is a great achievement in this study. In order to reinforce this study, it needs further compensative research, including cost factors which could not be applied to modeling the subject ports by lack of consistently qualified in ASEAN.
Park, Moo-Yong;Sagong, Dong-Hoon;Kweon, Hun-Joong;Do, Yun-Su;Song, Yang-Yik;Lee, Dong-Hyuk
Horticultural Science & Technology
/
v.31
no.5
/
pp.523-530
/
2013
This study investigated the influence of seasonal incidence and defoliation degree caused by Marssonina blotch (Diplocarpon mali Harada et Sawamura) on shoot growth and fruit quality of 'Fuji'/M.9 apple tree. The occurrence of marssonina blotch in Gunwi region was observed from the mid of July 2009, and percentage of defoliation was about 10% in the end of October. In 2010, that started in the early of June. By the end of September, percentage of defoliation was 20% and it reached 50% in late October. Secondary growth of terminal shoot in 2010 was three times as much as that in 2009. In 2009, the soluble solid contents and the red color (hunter a value) during fruit maturation increased to $13.8^{\circ}Brix$ and 16.2, respectively. In 2010, the soluble solid content during fruit maturation remained in the $12.1-12.6^{\circ}Brix$ range after early October, and the red color began to increase after the end of September when the percentage of defoliation was 20%. As for fruit quality by defoliation degree, the red color decreased when defoliation percentage of bourse shoot at the end of October was more than 30%. Fruit weight and soluble solids also decreased when defoliation percentage was more than 50%. Fruit length and diameter of over 30% defoliation treatments begun to decrease after the middle of August, compared with those of under 30% defoliation treatment. Photosynthetic rate of the leaves that was located at the secondary growth of bourse shoot was similar to that of leaves which was located at the middle of bourse shoot. However, the size and the soluble solid content of fruit were not affected by photosynthesis activities of bourse shoot leaves which was develop in the secondary shoot growth.
Jeong, Young Joo;Park, Sung Woo;Seo, Mi Kyoung;Kim, Sang-Jin;Lee, Won Hee;Kim, Mooseong;Urm, Sang-Hwa;Lee, Jung Goo;Seog, Dae-Hyun
Journal of Life Science
/
v.31
no.3
/
pp.257-265
/
2021
Heterotrimeric kinesin-2 is a molecular motor protein of the kinesin superfamily (KIF) that moves along a microtubule plus-end directed motor protein. It consists of three different motor subunits (KIF3A, KIF3B, and KIF3C) and a kinesin-associated protein 3 (KAP3) that form a heterotrimeric complex. Heterotrimeric kinesin-2 interacts with many different binding proteins through the cargo-binding domain of the KIF3s. The activity of heterotrimeric kinesin-2 is regulated to ensure that the cargo is directed to the right place at the right time. How this regulation occurs, however, remains in question. To identify the regulatory proteins for heterotrimeric kinesin-2, we performed yeast two-hybrid screening and found a specific interaction with creatine kinase B (CKB), which is the brain isoform of cytosolic creatine kinase enzyme. CKB bound to the cargo-binding domain of KIF3A but did not interact with the KIF3B, KIF5B, or KAP3 in the yeast two-hybrid assay. The carboxyl (C)-terminal region of CKB is essential for the interaction with KIF3A. Another protein kinase, CaMKIIa, interacted with KIF3A, but GSK3a did not interact with KIF3A in the yeast two-hybrid assay. KIF3A interacted with GST-CKB-C but not with GSK-CKB-N or GST alone. When co-expressed in HEK-293T cells, CKB co-localized with KIF3A and co-immunoprecipitated with KIF3A and KIF3B but not KIF5B. These results suggest that the CKB-KIF3A interaction may regulate the cargo transport of heterotrimeric kinesin-2 under energy-compromised conditions in cells.
Jeongeup, Jeollabuk-do, is an area that requires attention from those who study the history of Korean thought. In addition, Jeongeup is an area wherein many works were recorded for the first time in literary history. This is the case with Jeongeupsa as a style of Baekje songs and the lyrics of the noble families of the Joseon Dynasty, Sangchungok. Jeongeup is likewise the location where Choi Chi-won (857~?) was selected to serve as a local taesu (viceroy) and where a unique tradition of music and style were passed down. In this paper, the relationship between Choi Chi-won's role in the process of establishing a silent Confucian academy in Jeongeup and the emergence of scholar culture was examined. When Choi Chi-won left after his term in office, a birth shrine called Taesansa Temple was built to repay the selection of the villagers, and it became the source that led to the opening of the Confucian academy Museongseowon in the future. Jeongeup will be shown to be the location where Choi Chi-won realized his aspirations and honed his capabilities. In particular, Choi Chi-won's played a crucial role in the mid-Joseon Dynasty by supporting the construction and securing the name of Museongseowon. That is why Choi Chi-won was able to be revived as a symbolic figure in the region. In addition, it can be seen that the shape of Choi Chi-won was more sedentary- in the form of a Confucian scholar- and Confucian scholars emphasized the transfer of portraits at Museongseowon. Through the poetry written by Choi Chi-won, readers can learn about the worries and perceptions of scholars during those times. Although his value in the field of poetry is diverse, he can especially be recognized as a Confucian intellectual. In a large number of his works, he expresses his anxiety, agony, and critical inner consciousness all of which came from his encounter with the realities of his time. In fact, Choi Chi-won showed his qualities as a prominent literary figure of his time who had extraordinary aspirations and an admirable work ethic. However, he failed to overcome his regional and mental alienation as a poet in neighboring countries. Therefore, he internalized a sort of fierceness in terms of his perception of the world. However, it seems that it was rather a factor that made his work exhibit a strong lyrical style. In addition, Choi Chi-won's collection of writings includes a number of works that strongly criticized various forms of pathological phenomena caused by terminal phenomena of the time. He also highlighted the wrong in society by realistically depicting the lives poor and needy people and their eventual sacrifice via distorted relationships. This can be read encapsulating the agony of intellectuals of that time. The dictionary definition of a 'Confucian scholar' is "a Confucian term referring to a person or class that embodies Confucian ideology," and in its contemporary meaning it suggests " ⋯ an example of a personality, but not an identity, and the conscience of one's time period as a source of human morality inwardly and social order outwardly." In this respect, it could even be said that Choi Chi-won could be considered the originator of scholar culture.
Youngmin Kim;Seon Woong Jang ;Heung-Min Kim;Tak-Young Kim;Suho Bak
Korean Journal of Remote Sensing
/
v.39
no.5_1
/
pp.589-597
/
2023
Floating debris in large quantities from land during heavy rainfall has adverse social, economic, and environmental impacts, but the monitoring system for the concentration area and amount is insufficient. In this study, we proposed an efficient monitoring method for floating debris entering the river during heavy rainfall in Daecheong Lake, the largest water supply source in the central region, and applied remote sensing techniques to estimate the standing-stock of floating debris. To investigate the status of floating debris in the upper of Daecheong Lake, we used a tracking buoy equipped with a low-orbit satellite communication terminal to identify the movement route and behavior characteristics, and used a drone to estimate the potential concentration area and standing-stock of floating debris. The location tracking buoys moved rapidly during the period when the cumulative rainfall for 3 days increased by more than 200 to 300 mm. In the case of Hotan Bridge, which showed the longest distance, it moved about 72.8 km for one day, and the maximum moving speed at this time was 5.71 km/h. As a result of calculating the standing-stock of floating debris using a drone after heavy rainfall, it was found to be 658.8 to 9,165.4 tons, with the largest amount occurring in the Seokhori area. In this study, we were able to identify the main concentrations of floating debris by using location-tracking buoys and drones. It is believed that remote sensing-based monitoring methods, which are more mobile and quicker than traditional monitoring methods, can contribute to reducing the cost of collecting and processing large amounts of floating debris that flows in during heavy rain periods in the future.
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