Subolesin (4D8), the ortholog of insect akirins, is a highly conserved protective antigen and thus has the potential for development of a broad-spectrum vaccine against ticks and mosquitoes. To date, no protective antigens have been characterized nor tested as candidate vaccines against Dermacentor silvarum bites and transmission of associated pathogens. In this study, we cloned the open reading frame (ORF) of D. silvarum 4D8 cDNA (Ds4D8), which consisted of 498 bp encoding 165 amino acid residues. The results of sequence alignments and phylogenetic analysis demonstrated that D. silvarum 4D8 (Ds4D8) is highly conserved showing more than 81% identity of amino acid sequences with those of other hard ticks. Additionally, Ds4D8 containing restriction sites was ligated into the pET-32(a+) expression vector and the recombinant plasmid was transformed into Escherichia coli rosetta. The recombinant Ds4D8 (rDs4D8) was induced by isopropyl ${\beta}$-D-thiogalactopyranoside (IPTG) and purified using Ni affinity chromatography. The SDS-PAGE results showed that the molecular weight of rDs4D8 was 40 kDa, which was consistent with the expected molecular mass considering 22 kDa histidine-tagged thioredoxin (TRX) protein from the expression vector. Western blot results showed that rabbit anti-D. silvarum serum recognized the expressed rDs4D8, suggesting an immune response against rDs4D8. These results provided the basis for developing a candidate vaccine against D. silvarum ticks and transmission of associated pathogens.
By SDS-polyacrylamide gel elcctrophoresis (SDS-PAGE) and immunoblot analysis that a protein with 66,000 daltons in size was recognized by P. aeruginosa anti-exotoxin A from P. sp. PY002. The yields of exotoxin A in P. sp. PY002 culture were influenced by the concentration of iron in the culture media. Increasing of the exotoxin A production and siderophore production was made slight increasing in the MKB medium. On the other hand, to clone the gene encoding the exoloxin A genomic library of P. sp. PY002 was constructed in pBluescript SK(+). From this library a exotoxin A homologous gene was isolated by immunological hybridization method using P. aemginosa anti-exoloxin A as a probe. Two putative clones were isolated and designated pETA23 and pETA42. Thc restriction analysis ol pETA42 demonstrated that thc 1760 bp insert contained one NcoI, PvuII, SstI, Kpnl and EcoRI site and three SmaI and HaeD sites.
Watermelon production is often limited by powdery mildew in areas with a large daily temperature range. Development of resistant watermelon cultivars can protect against powdery mildew; however, little is known about the characteristics of its causal agents. Here, we identified the genus and race of a causal pathogen of powdery mildew in Ansung province of South Korea, and developed molecular markers for the generation of resistant watermelon cultivars. The causal pathogen was determined to be Podosphaera xanthii based on multiple sequence alignments of internal transcribed spacers (ITS) of rDNA. The physiological race was identified as 1W, and the Ansung isolate was named P. xanthii 1W-AN. Following inoculation with the identified P. xanthii 1W-AN, we found inheritance of the resistant gene fitting a single dominant Mendelian model in a segregated population ('SBA' ${\times}$ PI 254744). To develop molecular markers linked to fungus-resistant loci, random amplified polymorphic DNA (RAPD) was accomplished between DNA pooled from eight near-isogenic lines (NILs; $BC_4F_6$), originated from PI 254744 and susceptible 'SBB' watermelon. After sequencing bands from RAPD were identified in all eight NILs and PI254744, 42 sequence-characterized amplifiedregion (SCAR) markers were developed. Overall, 107 $F_2$ plants derived from $BC_4F_6$ NIL-1 ${\times}$ 'SBB' were tested, and one SCAR marker was selected. Sequence comparison between the SCAR marker and the reference watermelon genome identified three Nco I restriction enzyme sites harboring a single nucleotide polymorphism, and codominant cleavage-amplified polymorphic site markers were subsequently developed. A CAPS marker was converted to a high-resolution melt (HRM) marker, which can discriminate C/T SNP (254PMR-HRM3). The 254PMR-HRM3 marker was evaluated in 138 $F_{2:3}$ plants of a segregating population ('SBA' ${\times}$ PI254744) and was presumed to be 4.3 cM from the resistance locus. These results could ensure P. xanthii 1W-AN resistance in watermelon germplasm and aid watermelon cultivar development in marker-assist breeding programs.
Journal of the Institute of Electronics and Information Engineers
/
v.50
no.11
/
pp.180-187
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2013
The dinger in solar generation is the amount of sunlight which the solar cells absorb. Various types of solar trackers, which rotate themselves in order to make the solar cells face the sun as much as possible, have been developed, and especially the method of tracking with two axes has greatly contributed in increasing the generation amount at work sites. Among theses 2-axis solar trackers, the inclination-dependent 2-axis solar tracker are widely utilized for its advantages of requiring less initial investment and easy maintenance due to a solid structure. However, the drawback is that the generation efficiency is relatively low because of the structural restriction that limits the rotation angle, thus making it less efficient when tracking the sun. This paper proposes a method to increase the generation efficiency of the inclination-dependent 2-axis solar tracker. It also contains the derived equations needed for precise controlling along with a method to keep tracking with the other axis even when one has reached its angle limit. To confirm that the proposed method increases the amount of incidence onto the solar cells, formulas needed for operation on the proposed method and tracking the exact position of the sun are derived, and applying this to the quarterly data of Korea Astronomy and Space Science Institute it shows maximum over 11.1% more incidence compared to existing methods.
The matrix metalloproteinase (MMP) family plays essential roles in physiological processes such as embryonic development, angiogenesis, wound healing, and tissue homeostasis as a consequence of MMPr capacity for breaking down many types of extracellular matrix proteins. Imbalanced regulation of MMP expression can also lead to pathological conditions such as tumor progression. We recently reported that the Drosophila Mmp1 gene is highly expressed in the digestive tract and is required for the maintenance of intestinal homeostasis such as by restriction of uncontrolled intestinal stem cell proliferation. However, the regulatory mechanisms of MMP gene expression in the intestine remain unclear. In this study, we determined that the expression of Mmp1 is regulated by the homeodomain transcription factor Caudal. Experiments using the targeted expression of Caudal under the regulation of Gal4-UAS system indicated that endogenous Caudal is required for the Mmp1 gene expression in the adult Drosophila intestine and that exogenous Caudal induces Mmp1 expression. Transient transfection experiments indicated that Caudal can activate the promoter activity of Mmp1 and that several putative Caudal binding sites in the 5'-flanking region of the Mmp1 gene may be critical to the upregulation by Caudal. Our data suggest that Mmp1 is one of the target genes of Caudal in physiological normal condition and in tumorigenesis.
Morphological observation of roots and molecular technique were used to investigate the symbiotic relationships between arbuscular mycorrhizal (AM) fungi and ginseng roots. Korean ginseng, Panax ginseng, was collected from 8 sites in Korea. Colonization pattern of AM fungi in ginseng roots was determined as an Arum type under light microscopes. Nested PCR using AM fungal specific primers was employed to amplify a partial region on 18s rDNA of AM fungi from the root extracted mixed DNA. The amplified DNA was cloned and analyzed by random fragment length polymorphism (RFLP) with restriction enzymes, AluI, HinfI and AsuC21. One from each RFLP pattern was selected for sequencing. A total 16 clones were sequenced and identified as 2 species of AM fungi; Paraglomus brasilianum and Glomus spurcum. Paramglomus brasilianum was found from most of the ginseng roots, in this syudy suggesting that this species of AM fungi could have specific relationship with the ginseng root. Possible roles of AM fungal species in ginseng roots are discussed.
Journal of the Korean Institute of Traditional Landscape Architecture
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v.29
no.2
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pp.108-117
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2011
In drafting the standards on changes in current conditions, the height or the number of stories is applied in a restrictive manner by limiting to securing the distance to vertical surface for cultural properties within the influence investigation area, but this is expected to have a negative impact on the surrounding sceneries as well as results in the dwarfing phenomenon for precious cultural properties. That is, the preparation for supplementing the insufficient objectivity that is likely to take place during the process of drafting the standards on changes in current conditions. Thus the author attempts to suggest the analytic method for the decision making related to objective and reasonable determination and regulation of the changes in current conditions through computer based simulation work that considers the cultural properties and surrounding environments under investigation. In order to achieve such research objectives, the author reviewed the subject sites where the cultural property dwarfing phenomenon was expected to occur in case of the permission for the changes in current conditions or where the impact of natural landscape and natural feature on the earth is less than architectural building or artificial structure or where the new policy program is likely to be adopted due to incomplete establishment of current condition change standard within influence investigation area, among other cultural properties with architectural building or artificial structure nearby located in Cheonan city and then selected Cheonansaji Dangganjiju(flag poles) and Jiksanhyun Gwana(government office). The author then undertook the quantitative visibility analysis in order to determine the comprehensive prospect rights for the cultural properties and surrounding environments concerned.
Electronic commerce, commonly known as e-commerce or eCommerce, has become a major business trend in these days. The amount of trade conducted electronically has grown extraordinarily by developing the Internet technology. Most electronic commerce has being conducted between businesses to customers; therefore, the researches with respect to e-commerce are to find customer's needs, behaviors through statistical methods. However, the statistical researches, mostly based on a questionnaire, are the static researches, They can tell us the dynamic relationships between initial purchasing and repurchasing. Therefore, this study proposes dynamic research model for analyzing the cause of initial purchasing and repurchasing. This paper is based on the System-Dynamic theory, using the powerful simulation model with some restriction, The restrictions are based on the theory TAM(Technology Acceptance Model), PAM, and TPB(Theory of Planned Behavior). This article investigates not only the customer's purchasing and repurchasing behavior by passing of time but also the interactive effects to one another. This research model has six scenarios and three steps for analyzing customer behaviors. The first step is the research of purchasing situations. The second step is the research of repurchasing situations. Finally, the third step is to study the relationship between initial purchasing and repurchasing. The purpose of six scenarios is to find the customer's purchasing patterns according to the environmental changes. We set six variables in these scenarios by (1) changing the number of products; (2) changing the number of contents in on-line shopping malls; (3) having multimedia files or not in the shopping mall web sites; (4) grading on-line communities; (5) changing the qualities of products; (6) changing the customer's degree of confidence on products. First three variables are applied to study customer's purchasing behavior, and the other variables are applied to repurchasing behavior study. Through the simulation study, this paper presents some inter-relational result about customer purchasing behaviors, For example, Active community actions are not the increasing factor of purchasing but the increasing factor of word of mouth effect, Additionally. The higher products' quality, the more word of mouth effects increase. The number of products and contents on the web sites have same influence on people's buying behaviors. All simulation methods in this paper is not only display the result of each scenario but also find how to affect each other. Hence, electronic commerce firm can make more realistic marketing strategy about consumer behavior through this dynamic simulation research. Moreover, dynamic analysis method can predict the results which help the decision of marketing strategy by using the time-line graph. Consequently, this dynamic simulation analysis could be a useful research model to make firm's competitive advantage. However, this simulation model needs more further study. With respect to reality, this simulation model has some limitations. There are some missing factors which affect customer's buying behaviors in this model. The first missing factor is the customer's degree of recognition of brands. The second factor is the degree of customer satisfaction. The third factor is the power of word of mouth in the specific region. Generally, word of mouth affects significantly on a region's culture, even people's buying behaviors. The last missing factor is the user interface environment in the internet or other on-line shopping tools. In order to get more realistic result, these factors might be essential matters to make better research in the future studies.
Zoysiagrasses are important turf plants used for school playgrounds, parks, golf courses, and sports fields. The two most popular zoysiagrass species are Zoysia japonica and Zoysia sinica. These are widely distributed across different growing zones and are morphologically distinguishable from each other; however, it is phenotypically difficult to differentiate those that grow along the coastal line from those in beach area habitats. A combination of morphological and molecular approaches is desirable to efficiently identify these two plant cultivars. In this study, we used a rapid identification system based on DNA barcoding of the nrDNA-internal transcribed spacer (ITS) regions. The nrDNA-ITS regions of ITS1, 5.8S nrDNA, and ITS2 from Z. japonica, Z. sinica, Agrostis stolonifera, and Poa pratensis were DNA barcoded to classify these grasses according to their molecular identities. The nrDNA-ITS sequences of these species were found at 686 bp, 687 bp, 683 bp, and 681 bp, respectively. The size of ITS1 ranged from 248 to 249 bp, while ITS2 ranged from 270 to 274 bp. The 5.8S coding region ranged from 163 - 164bp. Between Z. japonica and Z. sinica, nineteen (2.8%) nucleotide sites were variable, and the G+C content of the ITS region ranged from 55.4 to 63.3%. Substitutions and insert/deletion (indel) sites in the nrDNA-ITS sequence of Z. japonica and Z. sinica were converted to cleaved amplified polymorphic sequence (CAPS) markers, and applied to the Zoysia grasses sampled to verify the presence of these markers. Among the 62 control and collected grass samples, we classified three groups: 36 Z. japonica, 22 Z. sinica, and 4 Z. japonica/Z. sinica hybrids. Morphological classification revealed only two groups; Z. japonica and Z. sinica. Our results suggest that used of the nrDNA-ITS barcode region and CAPS markers can be used to distinguish between Z. japonica and Z. sinica at the species level.
Han, Seung Hyun;Kim, Jung Hwan;Kang, Won Seok;Hwang, Jae Hong;Park, Ki Hyung;Kim, Chan-Beom
Korean Journal of Environment and Ecology
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v.33
no.4
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pp.453-461
/
2019
This study was conducted to monitor the soil characteristics and growth of Pinus densiflora and to determine the effect of soil characteristics on growth rate five years after an ecological restoration project in Baekdudaegan ridge including Ihwaryeong, Yuksimnyeong, and Beoljae sites. The ecological restoration project was executed with the forest of P. densiflora in 2012-2013. In April 2018, we collected soil samples from each site and measured the height and the diameter at breach height (DBH) of P. densiflora. Although there was no significant change of soil pH compared to the early stage of restoration (one year after the project), it was high in Ihwaryeong, and Beoljae with values of 7.7 and 6.4, respectively. Also, the organic matter decreased by 70-80%, and the available phosphorus (P) was unchanged in three restoration sites. The decreased organic matter can be attributed to restriction of inflow and thus decomposition of litter in the early stage after the restoration. The tree height growth rate ($m\;yr^{-1}$) of P. densiflora in Yuksimnyeong was the highest at 1.02, followed by Beolja at 0.75 and Ihwaryeong at 0.17. The height growth rate showed negative relationships with soil pH and cations, including Na and Ca concentrations and a positive relationship with available phosphate. The low growth rate in the Ihwaryeong site, in particular, might result from the poor nutrient availability due to high soil pH and the decrease in water absorption into the root due to high Na and Ca concentrations. The substantial reduction of organic matter after five years indicates that the need for soil improvement using chemical fertilizer and biochar.
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