• Title/Summary/Keyword: Vector species

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Effective Combination of Agrobacterium tumefaciens Strains and Ti Plasmids for the Construction of Plant Vector System

  • Kim, Mi-Suk;Park, Jeong-Du;Eum, Jin-Seong;Sim, Woong-Seop
    • Journal of Plant Biology
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    • v.39 no.3
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    • pp.179-184
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    • 1996
  • The purpose of this study is to obtain the most efficient combination of Agrobacterium tumefaciens strains and Ti plasmids for the construction of dicotyledonous plant vector system. Ti plasmid-curing A. tumefaciens A136 and KU12C3 were transformed with four kinds of Ti plasmids, pTiBo542, pTiA6, pTiKU12 and pTiAch5, respectively. The stems of 28 species of dicotyledonous plants were then inoculated with these transformants and examined for crown gall formation. The different combination of A. tumefaciens strains and Ti plasmids showed quite a difference in terms of the crown gall formation. Agrobacterium strins A136 and KU12C3 have a same plant host range in case that both strains harour the same kind of Ti plasmid, pTiBo542 or pTiAch5. However, the above-mentioned both strains have quite different host range in the event of containing the same Ti plasmid, pTiKU12 or pTiA6. In case that KU12C3 contains pTiA6 or pTiKU12, this strain has a wider plant host range than A136. The plant host range of pTiBo542 is the widest, followed by pTiA6, pTiKU12 and pTiAch5. Twelve plants among 28 tested plants are not transformed by any virulent Agrobacterium strains used in this study. In conclusion, A. tumefaciens KU12C3 and A136 harboring pTiBo542 showed the widest host range for transforming dicotyledonous plants. Also, it was acertained that the host range of Ti plasmids is affected by chromosomal level.

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Cloning and Characterization of a Bile Salt Hydrolase from Enterococcus faecalis Strain Isolated from Healthy Elderly Volunteers (사람 분변에서 분리한 Enterococcusfaecalis가 생성하는 BileSaltHydrolase의 특징)

  • Eom, Seok-Jin;Kim, Geun-Bae
    • Journal of Dairy Science and Biotechnology
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    • v.29 no.1
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    • pp.49-54
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    • 2011
  • Bile salt hydrolase (BSH, EC 3.5.1.24) activity, which cleaves amide bond between carboxyl group (bile acid) and amino group (glycine or taurine), is commonly detected in gut-associated species of human and animal. During the screening of BSH active strains from the fecal samples of elderly human volunteers, strain CU30-2 was isolated on the basis of the highly active BSH producing activity. A bsh gene of the isolate was cloned into the pET22b expression vector and overexpressed in Escherichia coli BL21 (DE3) Gold by induction with 1mM IPTG. The overexpressed BSH enzyme with 6x His-tag was purified with apparent homogeneity using a $Ni^+$-NTA agarose column and characterized. The BSH enzyme of E. faecalis CU30-2 exhibited approximately 50 times higher activity against glycol-conjugated bile salts than tauro-conjugated bile salts having the highest activity against glycocholic acid. Considering the prevalence of E. faecalis strains in the human GI tract and glycol-conjugates dominated bile acid composition of human bile, further study is needed to investigate the impact of the BSH activity exerted by E. faecalis strains to the host as well as to the BSH producing strains.

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Transposon piggyBac mediated Ipax6 Expression in Malaria Vector Anopheles stephensi (말라리아 매개 모기 Anopheles stephensi에서 트랜스포존 piggyBac을 이용한 Pax6 발현)

  • Koo Hyeyoung
    • Development and Reproduction
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    • v.8 no.1
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    • pp.19-25
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    • 2004
  • Pax6, a member of the highly conserved homeobox gene family, is known to be expressed in spatially and temporally restricted pattern during embryogenesis. To examine the spatial expression pattern of Pax6 in malaria vector mosquito Anopheles stephemi, in different molecular environment, the germ line transformation technique using piggyBac transposon combined with the use of Pax6 specific 3xp3-EGFP marker was utilized. Four transgenic lines with a transformation rate of 6.7% were established. Transgenes were stably expressed in subsequent several generations. The transgenic lines showed 3 different expression pattern with spatial specificity, possibly due to enhancing and/or silencing position effects. In two transgenic lines, noble expression pattern of Pax6 was observed in the region that has not been previously reported in any animal species. The results show that the tranposon piggyBac mediated germ line transformation system can be used as an efficient tool for the generation of diverse spatially restricted reporter gene expression.

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Transfected HepG2 Cells for Evaluation of Catechin Effects on Alcohol-Induced CYP2E1 Cytotoxicity

  • LEE YOO-HYUN;HO JIN-NYOUNG;DONG MI-SOOK;PARK CHANG-HWAN;KIM HYE-KYUNG;HONG BUMSHIK;SHIN DONG-HOON;CHO HONG-YON
    • Journal of Microbiology and Biotechnology
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    • v.15 no.6
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    • pp.1310-1316
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    • 2005
  • To evaluate the toxicological properties of human cytochrome P450 2E1 (CYP2E1) induced by ethanol and possible protective effects of various green tea catechins on alcohol-induced toxicity, transfected HepG2 cells that stably and constitutively express human CYP2E1 were established using the recombinant retroviral expression vector. Exposure of the CYP2E1-expressing HepG2 cells to high concentration of ethanol (200 mM) for 5 days resulted in a more than $50\%$ increase of cytotoxicity, assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction, lactate dehydrogenase (LDH) release, and reactive oxygen species (ROS) production, and loss of normal morphology, in comparison with HepG2 cells containing control vector. Treatment of the cells with various catechins increased cell viability by more than 2-fold. (-)-Epicatechin gallate and(-)-catechin gallate at the lowest concentration ($5\;{\mu}M$) attenuated cell death induced CYP2E1 by $60-65\%$. Therefore, the results showed that the catechins, including epimerized catechins, have strong protective effects against alcohol-induced CYP2E1 toxicity, and it is correlated with antioxidant effect.

Comparison of Fine Grained Classification of Pet Images Using Image Processing and CNN (영상 처리와 CNN을 이용한 애완동물 영상 세부 분류 비교)

  • Kim, Jihae;Go, Jeonghwan;Kwon, Cheolhee
    • Journal of Broadcast Engineering
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    • v.26 no.2
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    • pp.175-183
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    • 2021
  • The study of the fine grained classification of images continues to develop, but the study of object recognition for animals with polymorphic properties is proceeding slowly. Using only pet images corresponding to dogs and cats, this paper aims to compare methods using image processing and methods using deep learning among methods of classifying species of animals, which are fine grained classifications. In this paper, Grab-cut algorithm is used for object segmentation by method using image processing, and method using Fisher Vector for image encoding is proposed. Other methods used deep learning, which has achieved good results in various fields through machine learning, and among them, Convolutional Neural Network (CNN), which showed outstanding performance in image recognition, and Tensorflow, an open-source-based deep learning framework provided by Google. For each method proposed, 37 kinds of pet images, a total of 7,390 pages, were tested to verify and compare their effects.

BACTERIAL IDENTIFICATION WITH RANDOM-CLONED RESTRICTION FRAGMENT OF Porphyromonas endodontalis ATCC 35406 GENOMIC DNA (무작위로 클로닝한 Porphyromonas endodontalis ATCC 35406 지놈 DNA의 제한절편 hybridization법에 의한 세균동정)

  • Um, Won-Seok;Han, Yoon-Soo
    • Restorative Dentistry and Endodontics
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    • v.20 no.2
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    • pp.645-654
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    • 1995
  • Porphyromonas endodontalis is a black-pigmented anaerobic Gram negative rod which is associated with endodontal infections. It has been isolated from infected dental root canals and submucous abscesses of endodontal origin. DNA probe is an available alternative, offering the direct detection of a specific microorganism. Nucleic-acid probes can be off different types: whole different: whole-genomic, cloned or oligonucleotide probes. Wholegenomic probes are the most sensitive because the entire genome is used for possible hybridization sites. However, as genetically similar species of bacteria are likely to be present in specimences, cross-reactions need to be considered. Cloned probes are isolated sequences of DNA that do not show cross-reactivity and are produced in quantity by cloning in a plasmid vector. Cloned probes can approach the sensitivity found with whole-genomic probes while avoiding known cross-reacting species. Porphyromonas endodontalis ATCC 35406 (serotype $O_1K_1$) was selected in this experiment to develop specific cloned DNA probes. EcoR I-digested genomic DNA fragments of P. endodontalis ATCC 35406 were cloned into pUC18 plasmid vector. From the E. coli transformed with the recombinant plasmid 4 clones were selected to be tested as specific DNA probes. Restriction-digested whole-genomic DNAs prepared from P. gingivalis 38(serotype a), W50(serotype b), A7A1-28(serotype C), P. intermedia 9336(serotype b), G8-9K-3(serotype C), P. endodontalis ATCC 35406(serotype $O_1K_1$), A. a Y4(serotype b), 75(serotype a), 67(serotype c), were each seperated on agarose gel electrophoresis, blotted on nylon membranes, and were hybridized with digoxigenin-dUTP labeled probe. The results were as follows: 1. Three clones of 1.6kb(probe e), 1.6kb(probe f), and 0.9kb(probe h) in size, were obtained. These clones were identified to be a part of the genomic DNA of P. endodontalis ATCC 35406 judging from their specific hybridization to the genomic DNA fragments of their own size on Southern blot. 2. The clones of 4.9kb(probe i) was identified to be a part of the genomic DNA of P. endodontalis ATCC 35406. but not to specific for itself. It was hybridized to P. gingivalis A7A1-28, P. intermedia G89K-3.

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Monitoring Culicine Mosquitoes (Diptera: Culicidae) as a Vector of Flavivirus in Incheon Metropolitan City and Hwaseong-Si, Gyeonggi-Do, Korea, during 2019

  • Bahk, Young Yil;Park, Seo Hye;Kim-Jeon, Myung-Deok;Oh, Sung-Suck;Jung, Haneul;Jun, Hojong;Kim, Kyung-Ae;Park, Jong Myong;Ahn, Seong Kyu;Lee, Jinyoung;Choi, Eun-Jeong;Moon, Bag-Sou;Gong, Young Woo;Kwon, Mun Ju;Kim, Tong-Soo
    • Parasites, Hosts and Diseases
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    • v.58 no.5
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    • pp.551-558
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    • 2020
  • The flaviviruses are small single-stranded RNA viruses that are typically transmitted by mosquitoes or tick vectors and are etiological agents of acute zoonotic infections. The viruses are found around the world and account for significant cases of human diseases. We investigated population of culicine mosquitoes in central region of Korean Peninsula, Incheon Metropolitan City and Hwaseong-si. Aedes vexans nipponii was the most frequently collected mosquitoes (56.5%), followed by Ochlerotatus dorsalis (23.6%), Anopheles spp. (10.9%), and Culex pipiens complex (5.9%). In rural regions of Hwaseong, Aedes vexans nipponii was the highest population (62.9%), followed by Ochlerotatus dorsalis (23.9%) and Anopheles spp. (12.0%). In another rural region of Incheon (habitat of migratory birds), Culex pipiens complex was the highest population (31.4%), followed by Ochlerotatus dorsalis (30.5%), and Aedes vexans vexans (27.5%). Culex pipiens complex was the predominant species in the urban region (84.7%). Culicine mosquitoes were identified at the species level, pooled up to 30 mosquitoes each, and tested for flaviviral RNA using the SYBR Green-based RT-PCR and confirmed by cDNA sequencing. Three of the assayed 2,683 pools (989 pools without Anopheles spp.) were positive for Culex flaviviruses, an insect-specific virus, from Culex pipiens pallens collected at the habitats for migratory birds in Incheon. The maximum likelihood estimation (the estimated number) for Culex pipiens pallens positive for Culex flavivirus was 25. Although viruses responsible for mosquito-borne diseases were not identified, we encourage intensified monitoring and long-term surveillance of both vector and viruses in the interest of global public health.

Regrowth Ability and Species Composition of Phytoplankton in International Commercial Ship's Ballast Water Berthed at Pusan and Daesan Ports (부산과 대산항에서 선박평형수에 유입된 식물플랑크톤의 종조성과 재성장능력)

  • Baek, Seung-Ho;Jang, Min-Chul;Shin, Kyoung-Soon
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.16 no.2
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    • pp.106-115
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    • 2011
  • The aim of this study is to assess the importance of ballast water discharge as a vector for the introduction of exotic species into Pusan and Daesan Ports, Korea. We also examined to understand the impacts of environmental factors on the survival success of introduced species by ship's ballast water in laboratory experiments. Seven ship's ballast water originated from the coastal water of China (Taicang, Ningbo and Jinshan), Japan (Tokuyama, Moji and Akita), and Singapore. According to PCA (principal components analysis) analysis, environmental factor in the each ballast and shipside waters were different by bioregion. Based on cluster analysis, the phytoplankton community structures were distinguished for ballast water origin. Most of the major taxonomic groups were diatoms and, the others were dinoflagellate, silcoflagellate and several fresh-waters species. In particular, species number and standing crops of phytoplankton in the ballast tanks decreased with the increasing age ofballast water(r = -0.35 for standing crop; r = -0.63 for species number). In the laboratory study, although phytoplankton in ballast water treatment did not survive even in optimal temperature, the in vivo fluorescence of phytoplankton viability increased under the nutrient typical of shipside water and F/2 medium at $15^{\circ}C$ and $20^{\circ}C$. The diatoms species such as Skeletonema costatum and Thalassiosira pseudonana in ballast water were successfully regrown. On the salinity gradient experiments for Shui Shan (2) vessel, several freshwater species, brackish and marine species were successfully adapted. Of these, S.costatum was able to tolerate a wide range of salinities (10 to 30 psu) and its species-specific viability was suitable for colonization.

Comparison of Cuticular Hydrocarbons of the Pine Sawyer (Monochamus saltuarius), Japanese Pine Sawyer (Monochamus alternatus) and Oak Longicorn Beetle (Moechotypa diphysis) (북방수염하늘소(Monochamus saltuarius), 솔수염하늘소(Monochamus alternatus), 털두꺼비하늘소(Moechotypa diphysis) 성충의 표피탄화수소 비교)

  • Lee, Jeong-Eun;Kim, Eun-Hee;Yoon, Chang-Mann;Kim, Gil-Hah
    • Korean journal of applied entomology
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    • v.49 no.3
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    • pp.211-218
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    • 2010
  • Cuticular hydrocarbons (CHCs) of the pine sawyer (Monochamus saltuarius), Japanese pine sawyer (M. alternatus) and oak longicorn beetle (Moechotypa diphysis) were analyzed by GC, GC-MS and compared. Monochamus beetles are typical vectors of pine wilt disease but Moechotypa diphysis, which belongs to the same family, is not. They possess different CHCs in carbon number: 23-25 in M. saltuarius, 25-32 in M. alternatus, and 23-29 in M. diphysis. In comparison to inter-species, these three species of adult beetles have different numbers and chains of constituents of CHCs. In comparison between male and female in intra-species, the quantities of CHCs show the difference but constituents are not. Major constituent of M. saltuarius were analyzed as n-pentacosane > n-nonacosane > n-heptacosane; those of M. alternatus were n-nonacosene > n-pentacosane > n-nonacosane; and those of M. diphysis were n-heptacosane > 13-methylheptacosane > 3-methylheptacosane. From the body surface, most saturated carbohydrates of 3 species beetles are composed of n-alkane (40.2 - 65.7%) and followed by olefines > monomethylalkanes that one or two double bonds in M. saltuarius and M. alternatus. Otherwise, M. diphysis have the difference in order of monomethylalkanes > olefins.

Identification of Tick Species Collected from Wild Boars and Habitats of Wild Boars and Domestic Pigs in the Republic of Korea

  • Chae, Jeong-Byoung;Kang, Jun-Gu;Kim, Heung-Chul;Chong, Sung-Tae;Lee, In-Yong;Shin, Nam-Shik;Chae, Joon-Seok
    • Parasites, Hosts and Diseases
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    • v.55 no.2
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    • pp.185-191
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    • 2017
  • Tick is one of the most important arthropods in the transmission of vector-borne diseases. In this study, we investigated the abundance and species of ticks associated with swine and their habitats to assess the risk of spread of tick-borne diseases in host species, such as wild boars. Ticks were collected from 24 grazing or traditionally reared domestic pig farms and 8 habitats of wild boars in 8 provinces and 1 city in the Republic of Korea, by using the dragging and flagging methods. Ticks were also collected directly from 49 wild boars by using fine forceps. A total of 9,846 hard ticks were collected, including 4,977 Haemaphysalis longicornis, 4,313 Haemaphysalis flava, 508 Ixodes nipponensis, 1 Ixodes turdus, and 47 Amblyomma testudinarium. A total of 240 hard ticks were collected from 49 wild boars, including 109 H. flava, 84 H. longicornis, and 47 A. testudinarium. A total of 578 hard ticks were collected from areas around domestic pig farms. Only 2 hard tick species, 546 H. longicornis and 32 H. flava, were collected from these areas. A total of 9,028 hard ticks were collected from wild boars of 8 habitats, including 4,347 H. longicornis, 4,172 H. flava, 508 I. nipponensis, and 1 I. turdus. A. testudinarium was collected only from wild boars, and I. nipponensis and I. turdus were collected only from the habitats of wild boars.