• Title/Summary/Keyword: Vector species

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Effect of Antibiotic Down-Regulatory Gene wblA Ortholog on Antifungal Polyene Production in Rare Actinomycetes Pseudonocardia autotrophica

  • Kim, Hye-Jin;Kim, Min-Kyung;Kim, Young-Woo;Kim, Eung-Soo
    • Journal of Microbiology and Biotechnology
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    • v.24 no.9
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    • pp.1226-1231
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    • 2014
  • The rare actinomycete Pseudonocardia autotrophica was previously shown to produce a solubility-improved toxicity-reduced novel polyene compound named $\underline{N}ystatin$-like $\underline{P}seudonocardia$ $\underline{P}olyene$ (NPP). The low productivity of NPP in P. autotrophica implies that its biosynthetic pathway is tightly regulated. In this study, $wblA_{pau}$ was isolated and identified as a novel negative regulatory gene for NPP production in P. autotrophica, which showed approximately 49% amino acid identity with a global antibiotic down-regulatory gene, wblA, identified from various Streptomycetes species. Although no significant difference in NPP production was observed between P. autotrophica harboring empty vector and the S. coelicolor wblA under its native promoter, approximately 12% less NPP was produced in P. autotrophica expressing the wblA gene under the strong constitutive $ermE^*$ promoter. Furthermore, disruption of the $wblA_{pau}$ gene from P. autotrophica resulted in an approximately 80% increase in NPP productivity. These results strongly suggest that identification and inactivation of the global antibiotic down-regulatory gene wblA ortholog are a critical strategy for improving secondary metabolite overproduction in not only Streptomyces but also non-Streptomyces rare actinomycete species.

Functional Expression of SAV3818, a Putative TetR-Family Transcriptional Regulatory Gene from Streptomyces avermitilis, Stimulates Antibiotic Production in Streptomyces Species

  • Duong, Cae Thi Phung;Lee, Han-Na;Choi, Si-Sun;Lee, Sang-Yup;Kim, Eung-Soo
    • Journal of Microbiology and Biotechnology
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    • v.19 no.2
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    • pp.136-139
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    • 2009
  • Avermectin and its analogs are major commercial antiparasitic agents in the fields of animal health, agriculture, and human infections. Previously, comparative transcriptome analysis between the low-producer S. avermitilis ATCC31267 and the high-producer S. avermitilis ATCC31780 using a S. avermitilis whole genome chip revealed that 50 genes were overexpressed at least two-fold higher in S. avermitilis ATCC31780. To verify the biological significance of some of the transcriptomics-guided targets, five putative regulatory genes were individually cloned under the strong-and-constitutive promoter of the Streptomyces expression vector pSE34, followed by the transformation into the low-producer S. avermitilis ATCC31267. Among the putative genes tested, three regulatory genes including SAV213, SAV3818, and SAV4023 exhibited stimulatory effects on avermectin production in S. avermitilis ATCC31267. Moreover, overexpression of SAV3818 also stimulated actinorhodin production in both S. coelicolor M145 and S. lividans TK21, implying that the SAV3818, a putative TetR-family transcriptional regulator, could be a global upregulator acting in antibiotic production in Streptomyces species.

Molecular Cloning of Estrogen Receptor $\alpha$ in the Masu Salmon, Oncorhynchus masou

  • Sohn, Young Chang
    • Journal of Aquaculture
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    • v.17 no.1
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    • pp.62-68
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    • 2004
  • A cDNA encoding the masu salmon, Oncorhynchus masou, estrogen receptor $\alpha$ (msER$\alpha$) was cloned from the pituitary gland by polymerase chain reaction (PCR). This cDNA contains an open reading frame encoding 513 amino acid residues, and the calculated molecular weight of this protein is about 56,430 Dalton. The amino acid sequences of the DNA binding and ligand binding domains of msER$\alpha$ showed high homology to those of other fish species (84-100%). Reverse transcription PCR analysis showed that the mRNA level of msER$\alpha$ in the pituitary was slightly higher in estradiol-17$\beta$(E2) injected masu salmon than that of control fish. To test the biological activity of msER$\alpha$, the cDNA was ligated to a mammalian expression vector and transfected into a gonadotrope-derived cell line, L$\beta$T2, with a reporter plasmid including estrogen responsive element. Expression of the reporter protein, luciferase, was E2 and msER$\alpha$-dependent. The masu salmon ER$\alpha$ is structurally conserved among teleost species and functions as a transcriptional activator in the pituitary cells.

Fast Leaf Recognition and Retrieval Using Multi-Scale Angular Description Method

  • Xu, Guoqing;Zhang, Shouxiang
    • Journal of Information Processing Systems
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    • v.16 no.5
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    • pp.1083-1094
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    • 2020
  • Recognizing plant species based on leaf images is challenging because of the large inter-class variation and inter-class similarities among different plant species. The effective extraction of leaf descriptors constitutes the most important problem in plant leaf recognition. In this paper, a multi-scale angular description method is proposed for fast and accurate leaf recognition and retrieval tasks. The proposed method uses a novel scale-generation rule to develop an angular description of leaf contours. It is parameter-free and can capture leaf features from coarse to fine at multiple scales. A fast Fourier transform is used to make the descriptor compact and is effective in matching samples. Both support vector machine and k-nearest neighbors are used to classify leaves. Leaf recognition and retrieval experiments were conducted on three challenging datasets, namely Swedish leaf, Flavia leaf, and ImageCLEF2012 leaf. The results are evaluated with the widely used standard metrics and compared with several state-of-the-art methods. The results and comparisons show that the proposed method not only requires a low computational time, but also achieves good recognition and retrieval accuracies on challenging datasets.

Isolation and Sequence Analysis of Two Ornithine Decarboxylase Antizyme Genes from Flounder (Paralichthys olivaceus)

  • LEE JAE HYUNG;SEO YONG BAE;YOON MOON YOUNG;CHOI JUNG DO;KIM YOUNG TAE
    • Journal of Microbiology and Biotechnology
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    • v.15 no.2
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    • pp.321-329
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    • 2005
  • Ornithine decarboxylase (ODC) antizyme is a key regulatory protein in the control of cellular polyamines. We have isolated two distinct ODC antizyme cDNA clones (AZS and AZL) from a flounder (Paralichthys olivaceus) brain cDNA library. Their sequences revealed that both clones required translational frameshifting for expression. Taking + 1 frameshifting into account, AZS and AZL products were 221 and 218 amino acid residues long, respectively, and shared $83.3\%$ amino acid sequence identity. Comparison of the structure and nucleotide sequence of the antizyme genes showed that the genes were highly conserved in flounder, zebrafish, mouse, and human. A phylogenetic tree was constructed, based on the antizyme amino acid sequences from various species. The presence of the two types of antizyme mRNA species in brain, kidney, liver, and embryo was confirmed by using the reverse transcription­polymerase chain reaction (RT-PCR) and Northern blot analysis. Recombinant proteins of flounder ODC antizymes, containing His-Nus-S tag at the amino-terminus, were overexpressed as His-AZL and His-AZS fusion proteins in Escherichia coli BL21 (DE3) pLys by using the pET­44a(+) expression vector.

Ancient Human Parasites in Ethnic Chinese Populations

  • Yeh, Hui-Yuan;Mitchell, Piers D.
    • Parasites, Hosts and Diseases
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    • v.54 no.5
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    • pp.565-572
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    • 2016
  • Whilst archaeological evidence for many aspects of life in ancient China is well studied, there has been much less interest in ancient infectious diseases, such as intestinal parasites in past Chinese populations. Here, we bring together evidence from mummies, ancient latrines, and pelvic soil from burials, dating from the Neolithic Period to the Qing Dynasty, in order to better understand the health of the past inhabitants of China and the diseases endemic in the region. Seven species of intestinal parasite have been identified, namely roundworm, whipworm, Chinese liver fluke, oriental schistosome, pinworm, Taenia sp. tapeworm, and the intestinal fluke Fasciolopsis buski. It was found that in the past, roundworm, whipworm, and Chinese liver fluke appear to have been much more common than the other species. While roundworm and whipworm remained common into the late 20th century, Chinese liver fluke seems to have undergone a marked decline in its prevalence over time. The iconic transport route known as the Silk Road has been shown to have acted as a vector for the transmission of ancient diseases, highlighted by the discovery of Chinese liver fluke in a 2,000 year-old relay station in northwest China, 1,500 km outside its endemic range.

On the Rat-Fleas in Ducksom and Kwangnaru Areas of Seoul City (서울시 뚝섬, 광나루 일대의 쥐벼룩에 대하여)

  • CHU, In-Ho;HONG, Seun-Mo
    • The Korean Journal of Zoology
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    • v.1 no.1
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    • pp.1-6
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    • 1958
  • A rat-fleas survey was carried out in Ducksonm and Kwangnaru areas of Seoul city from Janurary to December 1955 . In this survey, 1,204 redents were trapped alive and 740 of which being Rattus norvegicus, 457 Apodemus agrarius, 5 Eutamiias asiaticus and 2 shrewes ((crocidura rusila). On these rodents 270 fleas were collected and identified thus : Ceratopylidae : Monopsyllus anisus (Rothschild 1908) ···200 Leptosylla segnis(Schonherr, 1832)···1 Hystrichopsyllidae : Ctenophthalmus congener, Rothschild ···43 Neopsylla bidentiformis (Wagner, 1893)···11 Stenoponia sidimi Marikovsky, 1936···9 Pulcidae : Xenopsylla cheopis (Rothschild, 1903)···6 On the average 8.5% of the rodents were infestated with fleas. The infestation rate showed a marked varying degree in a year and three high incidence peaks in Spring (10.3%), Summer (11.4%-11.8%) and Winter (13.7%). The average number of fleas per rat (flea-index) was 0.22 for the whole year with a maximum of 0.41 in December and a minimum of 0.03 in February. Of all, the most prevalent species are Monopsyllus anisus (74.1%) and secondly Ctenophthalmus congener (15.9%). both species occupied 90% of all. Xenopsylla cheopis, a known vector of plague, was found very rare in our survey and only appeared in Winter. The striking lower incidence of flea-index compared with previous data (6.6 by Nagahana, 1931-1936) appears to be due to the nationwide application of DDT and BHC insecticides since 1945.

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Molecular Authentication of Schisandrae Fructus and Analysis of Phylogenetic Relationship based on nrDNA-ITS sequences (nrDNA-ITS 분자마커를 이용한 오미자(五味子) 종 감별 및 기원분석 -ITS 염기서열을 이용한 오미자(五味子) 감별-)

  • Moon, Byeong-Cheol;Ji, Yun-Ui;Seo, Hyeong-Seok;Lee, A-Young;Chun, Jin-Mi;Kim, Ho-Kyoung
    • The Korea Journal of Herbology
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    • v.25 no.4
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    • pp.47-54
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    • 2010
  • Objectives : The original plant species of Schisandrae Fructus (O-mi-ja) is prescribed as Schisandra chinensis $B_{AILL.}$, in Korea, but S. chinensis $B_{AILL.}$ and S. sphenanthera $R_{EHD.}$ et $W_{ILS.}$ in China. Moreover, fruit of several other species in genus Schisandra also have been used as the same herbal medicines. To develop a reliable method for correct identification of Schisandrae Fructus and to evaluate the phylogenetic relationship of S. chinensis and its related species, we analyzed internal transcribed spacer (ITS) sequences of nuclear ribosomal DNA (nrDNA). Methods : Twenty-four plant samples of three Schisandra species and one Kadsura species, S. chinensis $B_{AILL.}$, S. spenanthera $R_{EHD.}$ et $W_{ILS.}$, S. nigra $M_{ax.}$ and Kadsura japonica $D_{UNAL}$ were collected from each different native habitate and farm in Korea and China. The nrDNA-ITS region of each samples were amplified using ITS1 and ITS4 primer and nucleotide sequences were determined after sub-cloning into the pGEM-Teasy vector. Authentic marker nucleotides were estimated by the analysis of ClastalW based on the entire nrDNA-ITS sequence. Results : In comparative analysis of the nrDNA-ITS sequences, we found specific nucleotide sequences including indels (insertions and deletions) and substitutions to distinguish C. chinensis, S. spenanthera, S. nigra, and K. japonica. These sequence differences at corresponding positions are avaliable nucleotide markers to determine the botanical origin of O-mi-ja. Moreover, we evaluated the phylogenetic relationship of four plant species by the analysis of nrDNA-ITS sequences. Conclusions : These marker nucleotides would be useful to identify the official herbal medicines by the providing of definitive information that can identify each plant species and distinguish it from unauthentic adulterants for O-mi-ja.

Methods for sampling and analysis of marine microalgae in ship ballast tanks: a case study from Tampa Bay, Florida, USA

  • Garrett, Matthew J.;Wolny, Jennifer L.;Williams, B. James;Dirks, Michael D.;Brame, Julie A.;Richardson, R. William
    • ALGAE
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    • v.26 no.2
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    • pp.181-192
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    • 2011
  • Ballasting and deballasting of shipping vessels in foreign ports have been reported worldwide as a vector of introduction of non-native aquatic plants and animals. Recently, attention has turned to ballast water as a factor in the global increase of harmful algal blooms (HABs). Many species of microalgae, including harmful dinoflagellate species, can remain viable for months in dormant benthic stages (cysts) in ballast sediments. Over a period of four years, we surveyed ballast water and sediment of ships docked in two ports of Tampa Bay, Florida, USA. Sampling conditions encountered while sampling ballast water and sediments were vastly different between vessels. Since no single sample collection protocol could be applied, existing methods for sampling ballast were modified and new methods created to reduce time and labor necessary for the collection of high-quality, qualitative samples. Five methods were refined or developed, including one that allowed for a directed intake of water and sediments. From 63 samples, 1,633 dinoflagellate cysts and cyst-like cells were recovered. A native, cyst-forming, harmful dinoflagellate, Alexandrium balechii (Steidinger) F. J. R. Taylor, was collected, isolated, and cultured from the same vessel six months apart, indicating that ships exchanging ballast water in Tampa Bay have the potential to transport HAB species to other ports with similar ecologies, exposing them to non-native, potentially toxic blooms.

Catenotaenia dendritica (Cestoda: Catenotaeniidae) and Three Ectoparasite Species in the Red Squirrel, Sciurus vulgaris, from Cheongju, Korea

  • Choe, Seongjun;Lee, Dongmin;Park, Hansol;Jeon, Hyeong-Kyu;Lee, Youngsun;Na, Ki-Jeong;Lee, In-Yong;Eom, Keeseon S.
    • Parasites, Hosts and Diseases
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    • v.54 no.4
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    • pp.509-518
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    • 2016
  • Parasites are recorded from the red squirrel, Sciurus vulgaris, from Cheongju, the Republic of Korea. A total of 5 road-killed squirrels were thoroughly examined for internal and external parasites from November 2011 to May 2014. Total 4 parasite species, including 1 tapeworm and 3 ectoparasite species were recovered. They were morphologically identified as Catenotaenia dendritica (Cestoda: Catenotaeniidae), Hirstionyssus sciurinus, Leptotrombidium pallidum, and Ceratophyllus (Monopsyllus) indages. Among them, C. dendritica and H. sciurinus are recorded for the first time in the Korean parasite fauna. In addition, the possibility that the red squirrel could act as a reservoir host for a zoonotic disease like tsutsugamushi disease with L. pallidum as its vector has been raised.