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Development of Molecular Markers for Xanthomonas axonopodis Resistance in Soybean

  • Kim Ki-Seung;Van Kyujung;Kim Moon Young;Lee Suk-Ha
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.49 no.5
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    • pp.429-433
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    • 2004
  • A single recessive gene, rxp, controls the bacterial leaf pustule (BLP) resistance in soybean and in our previous article, it has been mapped on linkage group (LG) D2 of molecular genetic map of soybean. A total of 130 recombinant inbred lines (RILs) from a cross between BLP-resistant SS2-2 and BLP-susceptible Jangyeobkong were used to identify molecular markers linked to rxp. Fifteen simple sequence repeat (SSR) markers on LG D2 were screened to construct a genetic map of rxp locus. Only four SSR markers, Satt135, Satt372, Satt448, and Satt486, showed parental polymorphisms. Using these markers, genetic scaffold map was constructed covering 26.2cM. Based on the single analysis of variance, Satt372 among these four SSR markers was the most significantly associated with the resistance to BLP. To develop new amplified fragment length polymorphism (AFLP) marker linked to the resistance gene, bulked segregant analysis (BSA) was employed. Resistance and susceptible bulks were made by pooling equal amount of genomic DNAs from ten of each in the segregating population. A total of 192 primer combinations were used to identify specific bands to the resistance, selecting three putative AFLP markers. These AFLP markers produced the fragment present in SS2-2 and the resistant bulk, and not in Jangyeobkong and the susceptible bulk. Linkage analysis revealed that McctEact97 $(P=0.0004,\;R^2=14.67\%)$ was more significant than Satt372, previously reported as the most closely linked marker.

Functional Implication of the tRNA Genes Encoded in the Chlorella Virus PBCV-l Genome

  • Lee, Da-Young;Graves, Michael V.;Van Etten, James L.;Choi, Tae-Jin
    • The Plant Pathology Journal
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    • v.21 no.4
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    • pp.334-342
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    • 2005
  • The prototype Chlorella virus PBCV-l encodes 11 tRNA genes and over 350 protein-encoding genes in its 330 kbp genome. Initial attempts to overexpress the recombinant A189/192R protein, a putative virus attachment protein, in E. coli strain BL21(DE3) SI were unsuccessful, and multiple protein bands were detected on Western blots. However, the full-length A189/192R recombinant protein or fragments derived from it were detected when they were expressed in E. coli BL21 CodonPlus (DE3) RIL, which contains extra tRNAs. Codon usage analysis of the a189/192r gene showed highly biased usage of the AGA and AVA codons compared to genes encoded by E. coli and Chlorella. In addition, there were biases of XXA/U($56\%$) and XXG/ C($44\%$) in the codons recognized by the viral tRNAs, which correspond to the codon usage bias in the PBCV-1 genome of XXA/U ($63\%$) over those ending in XXC/G ($37\%$). Analysis of the codon usage in the major capsid protein and DNA polymerase showed preferential usage of codons that can be recognized by the viral tRNAs. The Asn (AAC) and Lys (AAG) codons whose corresponding tRNA genes are duplicated in the tRNA gene cluster were the most abundant (i.e., preferred) codons in these two proteins. The tRNA genes encoded in the PBCV-l genome seem to play a very important role during the synthesis of viral proteins through supplementing the tRNAs that are frequently used in viral proteins, but are rare in the host cells. In addition, these tRNAs would help the virus to adapt to a wide range of hosts by providing tRNAs that are rare in the host cells.

Molecular analysis of genetic diversity, population structure, and phylogeny of wild and cultivated tulips (Tulipa L.) by genic microsatellites

  • Pourkhaloee, Ali;Khosh-Khui, Morteza;Arens, Paul;Salehi, Hassan;Razi, Hooman;Niazi, Ali;Afsharifar, Alireza;Tuyl, Jaap van
    • Horticulture, Environment, and Biotechnology : HEB
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    • v.59 no.6
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    • pp.875-888
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    • 2018
  • Tulip (Tulipa L.) is one of the most important ornamental geophytes in the world. Analysis of molecular variability of tulips is of great importance in conservation and parental lines selection in breeding programs. Of the 70 genic microsatellites, 15 highly polymorphic and reproducible markers were used to assess the genetic diversity, structure, and relationships among 280 individuals of 36 wild and cultivated tulip accessions from two countries: Iran and the Netherlands. The mean values of gene diversity and polymorphism information content were 0.69 and 0.66, respectively, which indicated the high discriminatory power of markers. The calculated genetic diversity parameters were found to be the highest in wild T. systola Stapf (Derak region). Bayesian model-based STRU CTU RE analysis detected five gene pools for 36 germplasms which corresponded with morphological observations and traditional classifications. Based on analysis of molecular variance, to conserve wild genetic resources in some geographical locations, sampling should be performed from distant locations to achieve high diversity. The unweighted pair group method with arithmetic mean dendrogram and principal component analysis plot indicated that among wild tulips, T. systola and T. micheliana Hoog exhibited the closest relationships with cultivated tulips. Thus, it can be assumed that wild tulips from Iran and perhaps other Middle East countries played a role in the origin of T. gesneriana, which is likely a tulip species hybrid of unclear origin. In conclusion, due to the high genetic variability of wild tulips, they can be used in tulip breeding programs as a source of useful alleles related to resistance against stresses.

Expression and Purification of Extracellular Solute-Binding Protein (ESBP) in Escherichia coli, the Extracellular Protein Derived from Bifidobacterium longum KACC 91563

  • Song, Minyu;Kim, Hyaekang;Kwak, Woori;Park, Won Seo;Yoo, Jayeon;Kang, Han Byul;Kim, Jin-Hyoung;Kang, Sun-Moon;Van Ba, Hoa;Kim, Bu-Min;Oh, Mi-Hwa;Kim, Heebal;Ham, Jun-Sang
    • Food Science of Animal Resources
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    • v.39 no.4
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    • pp.601-609
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    • 2019
  • Bifidobacterium longum KACC 91563 secretes family 5 extracellular solute-binding protein via extracellular vesicle. In our previous work, it was demonstrated that the protein effectively alleviated food allergy symptoms via mast cell specific apoptosis, and it has revealed a therapeutic potential of this protein in allergy treatment. In the present study, we cloned the gene encoding extracellular solute-binding protein of the strain into the histidine-tagged pET-28a(+) vector and transformed the resulting plasmid into the Escherichia coli strain BL21 (DE3). The histidine-tagged extracellular solute-binding protein expressed in the transformed cells was purified using Ni-NTA affinity column. To enhance the efficiency of the protein purification, three parameters were optimized; the host bacterial strain, the culturing and induction temperature, and the purification protocol. After the process, two liters of transformed culture produced 7.15 mg of the recombinant proteins. This is the first study describing the production of extracellular solute-binding protein of probiotic bacteria. Establishment of large-scale production strategy for the protein will further contribute to the development of functional foods and potential alternative treatments for allergies.

Breeding Hybrid Rice with Genes Resistant to Diseases and Insects Using Marker-Assisted Selection and Evaluation of Biological Assay

  • Kim, Me-Sun;Ouk, Sothea;Jung, Kuk-Hyun;Song, Yoohan;Le, Van Trang;Yang, Ju-Young;Cho, Yong-Gu
    • Plant Breeding and Biotechnology
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    • v.7 no.3
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    • pp.272-286
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    • 2019
  • Developing elite hybrid rice varieties is one important objective of rice breeding programs. Several genes related to male sterilities, restores, and pollinators have been identified through map-based gene cloning within natural variations of rice. These identified genes are good targets for introducing genetic traits in molecular breeding. This study was conducted to breed elite hybrid lines with major genes related to hybrid traits and disease/insect resistance in 240 genetic resources and F1 hybrid combinations of rice. Molecular markers were reset for three major hybrid genes (S5, Rf3, Rf4) and thirteen disease/insect resistant genes (rice bacterial blight resistance genes Xa3, Xa4, xa5, Xa7, xa13, Xa21; blast resistance genes Pita, Pib, Pi5, Pii; brown planthopper resistant genes Bph18(t) and tungro virus resistance gene tsv1). Genotypes were then analyzed using molecular marker-assisted selection (MAS). Biological assay was then performed at the Red River Delta region in Vietnam using eleven F1 hybrid combinations and two control vatieties. Results showed that nine F1 hybrid combinations were highly resistant to rice bacterial blight and blast. Finally, eight F1 hybrid rice varieties with resistance to disease/insect were selected from eleven F1 hybrid combinations. Their characteristics such as agricultural traits and yields were then investigated. These F1 hybrid rice varieties developed with major genes related to hybrid traits and disease/insect resistant genes could be useful for hybrid breeding programs to achieve high yield with biotic and abiotic resistance.

Copper Tolerance of Novel Rhodotorula sp. Yeast Isolated from Gold Mining Ore in Gia Lai, Vietnam

  • Kim Cuc Thi Nguyen;Phuc Hung Truong;Cuong Tu Ho;Cong Tuan Le;Khoa Dang Tran;Tien Long Nguyen;Manh Tuan Nguyen;Phu Van Nguyen
    • Mycobiology
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    • v.51 no.6
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    • pp.379-387
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    • 2023
  • In this study, twenty-five yeast strains were isolated from soil samples collected in the gold mining ore in Gia Lai, Vietnam. Among them, one isolate named GL1T could highly tolerate Cu2+ up to 10 mM, and the isolates could also grow in a wide range of pH (3-7), and temperature (10-40 ℃). Dried biomass of GL1 was able to remove Cu2+ effectively up to 90.49% with a maximal biosorption capacity of 18.1 mg/g at pH 6, temperature 30 ℃, and incubation time 60 min. Sequence analysis of rDNA indicated this strain was closely related to Rhodotorula mucilaginosa but with 1.53 and 3.46% nucleotide differences in the D1/D2 domain of the 28S rRNA gene and the ITS1-5.8S rRNA gene-ITS2 region sequence, respectively. Based on phylogenetic tree analysis and the biochemical characteristics, the strain appears to be a novel Rhodotorula species, and the name Rhodotorula aurum sp. nov. is proposed. This study provides us with more information about heavy metal-tolerant yeasts and it may produce a new tool for environmental control and metal recovery operations.

Marine-derived Ca-Mg complex influences lipid and glucose metabolism, serum metabolites, colostrum profile, and stress hormone in sows over four-parity periods

  • Sungbo Cho;Santi Devi Upadhaya;Woo Jeong Seok;Seyoung Mun;Haeun Lee;Rudolf H. van der Veen;Kyudong Han;In Ho Kim
    • Journal of Animal Science and Technology
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    • v.65 no.6
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    • pp.1308-1322
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    • 2023
  • Minerals is required small amounts among various nutrients, but it has a significant impact on sow longevity and reproduction performance. This study was carried out to see the beneficial effects of marine-derived Ca-Mg complex on the reproductive performance of sows during four-parity periods. Seventy-two gilts ([Yorkshire × Landrace] × Duroc), with an average body weight of 181 kg, were randomly allocated to three groups; CON (basal diet), 0.3LC (CON - MgO - 0.3% limestone + 0.4% Ca-Mg complex), and 0.7LC (CON - MgO - 0.7% limestone + 0.4% Ca-Mg complex). During parity 3 and 4, the expression level of SCD gene was lower in the umbilical cord of piglets born to 0.3LC and 0.7LC sows compared with the CON sows. During parity 2, 3 and 4, SLC2A2 and FABP4 gene expressions were higher in the umbilical cord of piglets born to 0.7LC sows and the placenta of sows from 0.3LC groups, respectively. Ca-Mg complex increased (p < 0.05) Ca and Mg concentrations in sows and their piglets' serum as well as in colostrum regardless of parities. The serum vitamin D concentration was higher (p < 0.05) in their first parity, whereas serum prolactin and estrogen concentrations were higher (p < 0.05) during the fourth and third parity, respectively. The growth hormone concentrations were higher (p < 0.05) in the piglets born to sows during the first and second parity. The fat and immunoglobulin A (IgA) concentrations in colostrum were higher (p < 0.05) during the third and fourth parity, respectively. A reduction (p < 0.05) in salivary cortisol, epinephrine, and norepinephrine concentrations was observed in 0.3LC and 0.7LC sow groups compared with CON after farrowing regardless of parity, however before farrowing, a reduction in norepinephrine was observed. Before farrowing, the epinephrine and norepinephrine concentrations were higher (p < 0.05) during the first and second parity. After farrowing, the concentration of these hormones was higher during the second parity. Taken together, sows' parity and dietary Ca-Mg complex supplementation influenced serum metabolites, colostrum nutrients, stress hormones as well as the gene expressions related to lipid and glucose metabolism.

Detection of Antibiotic Resistance and Resistance Genes in Enterococci Isolated from Sucuk, a Traditional Turkish Dry-Fermented Sausage

  • Demirgul, Furkan;Tuncer, Yasin
    • Food Science of Animal Resources
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    • v.37 no.5
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    • pp.670-681
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    • 2017
  • The aim of this study was to isolate enterococci in Sucuk, a traditional Turkish dry-fermented sausage and to analyze isolates for their biodiversity, antibiotic resistance patterns and the presence of some antibiotic resistance genes. A total of 60 enterococci strains were isolated from 20 sucuk samples manufactured without using a starter culture and they were identified as E. faecium (73.3%), E. faecalis (11.7%), E. hirae (8.3%), E. durans (3.3%), E. mundtii (1.7%) and E. thailandicus (1.7%). Most of the strains were found resistant to rifampin (51.67%) followed by ciprofloxacin (38.33%), nitrofurantoin (33.33%) and erythromycin (21.67%). All strains were found susceptible to ampicillin. Only E. faecium FYE4 and FYE60 strains displayed susceptibility to all antibiotics. Other strains showed different resistance patterns to antibiotics. E. faecalis was found more resistant to antibiotics than other species. Most of the strains (61.7%) displayed resistance from between two and eight antibiotics. The ermB, ermC, gyrA, tetM, tetL and vanA genes were detected in some strains. A lack of correlation between genotypic and phenotypic analysis for some strains was detected. The results of this study indicated that Sucuk manufactured without using a starter culture is a reservoir of multiple antibiotic resistant enterococci. Consequently, Sucuk is a potential reservoir for the transmission of antibiotic resistance genes from animals to humans.

An Information-Intensive Approach to the Molecular Pharmacology of Cancer

  • John N. Weinstein;Timothy G. Myers;Patrick M. O′Connor;Stephen H. Friend;Albert J. Fornace Jr;Kurt W. Kohn;Tito Fojo;Susan E. Bates;Lawrence V. Rubinstein;N. Leigh Anderson;John K. Buolamwini;Wiliam W. van Osdol;Anne P. Monks;Dominic A. Scudiero;Edward A. Sausville;Daniel W. Zaharevitz;Barry Bunow;Vellarkda N. Viswanadhan;Georage S. Johnson;Robert E. Wittes;Kennety D. Paull
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2001.08a
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    • pp.139-149
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    • 2001
  • Since 1990, the National Cancer Institute(NCI) has screened more than 60.000 compounds against a panel of 60 human cancer cell lines. The 50-percent growth-inhibitory concentration (GI$_{50}$) values encode unexpectedly rich, detailed information on mechanisms of drug action and drug resistance. Each compound's pattern is like a fingerprint, essentially unique among the many billions of distinguishable possibilities. These activity patterns are being used in conjunction with molecular structural features of the tested agents to explore the NCI's database of more than 460, 000 compounds, and they are providing insight into potential target molecules and modulators of activity in the 60 cell lines. For example, the information is being used to search for candidate anticancer drugs that are not dependent on intact p53 suppressor gene function for their activity. It remains to be seen how effective this information-intensive strategy will be at generating new clinically active agents.s.

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Effect of Phytolaccae Radix on the Proliferation of Transplanted-L1210 cells in Mice (상륙이 생쥐에 이식된 L1210 세포의 증식에 미치는 영향)

  • Han Mi Soak;Oh Chan Ha;Eun Jae Soon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.16 no.2
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    • pp.311-315
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    • 2002
  • Cellular death by apoptosis is an active process, depending on gene transcription and protein synthesis. It was reported that nitric oxide can induce apoptosis in several cancer cell-lines. We studied effects of Phytolacca esculentum van Houtt (Phytolaccaceae) Radix water extract (PRE) on the proliferation of transplanted-L1210 cells in mice. When PRE (500 mg/kg) was administered orally once a day for 7 days after transplantation of L1210 cells to mice, DNA fragmentation of transplanted-L1210 cells was induced and mitochondrial transmembrane potential of those cells was reduced. Additionally, DNA fragmentation of L1210 cells was induced by the treatment of PRE in vitro. Also, DNA fragmentation of L1210 cells was enhanced by co-culture with the peritoneal macrophages obtained from PRE-administered mice and was partly inhibited by L-NMMA in vitro. PRE enhanced the production of nitric oxide and tumor necrosis factor-α from peritoneal rnacrophages. These results suggest that PRE induces apoptosis of transplanted-L1210 cells via directive action on L1210 cells and stimulation of nitric oxide and tumor neaosis factor-α from macrophages.