• Title/Summary/Keyword: nod genes

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Flavonoid Biosynthesis: Biochemistry and Metabolic Engineering (Flavonoid 생합성:생화학과 대사공학적 응용)

  • Park, Jong-Sug;Kim, Jong-Bum;Kim, Kyung-Hwan;Ha, Sun-Hwa;Han, Bum-Soo;Kim, Yong-Hwan
    • Journal of Plant Biotechnology
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    • v.29 no.4
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    • pp.265-275
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    • 2002
  • Flavonoid biosynthesis is one of the most extensively studied areas in the secondary metabolism. Due to the study of flavonoid metabolism in diverse plant system, the pathways become the best characterized secondary metabolites and can be excellent targets for metabolic engineering. These flavonoid-derived secondary metabolites have been considerably divergent functional roles: floral pigment, anticancer, antiviral, antitoxin, and hepatoprotective. Three species have been significant for elucidating the flavonoid metabolism and isolating the genes controlling the flavonoid genes: maize (Zea mays), snapdragon (Antirrhinum majus) and petunia (Prtunia hybrida). Recently, many genes involved in biosynthesis of flavonoid have been isolated and characterized using mutation and recombinant DNA technologies including transposon tagging and T-DNA tagging which are novel approaches for the discovery of uncharacterized genes. Metabolic engineering of flavonoid biosynthesis was approached by sense or antisense manipulation of the genes related with flavonoid pathway, or by modified expression of regulatory genes. So, the use of a variety of experimental tools and metabolic engineering facilitated the characterization of the flavonoid metabolism. Here we review recent progresses in flavonoid metabolism: confirmation of genes, metabolic engineering, and applications in the industrial use.

CD7-Specific Single Chain Antibody Mediated Delivery of siRNA to T Cells Inhibits HIV Replication in a Humanized Mouse Model

  • Ban, Hong-Seok;Kumar, Priti;Kim, Na-Hyun;Choi, Chang-Son;Shankar, Premlata;Lee, Sang-Kyung
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2008.05a
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    • pp.62-64
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    • 2008
  • A major hurdle to the development of RNA interference as therapy for HIV infection is the delivery of siRNA to T lymphocytes which are difficult cells to transfect even in vitro. We have employed a single chain antibody to the pan T cell surface antigen CD7 was conjugated to an oligo-9-arginine peptide (scFvCD7-9R) for T cell-specific siRNA delivery in NOD/SCIDIL2${\gamma}$-/- mice reconstituted with human peripheral blood lymphocytes (Hu-PBL). Using a novel delivery, we first show that scFvCD7-9R efficiently delivered CD4 siRNA into human T cells in vitro. In vivo administration to Hu-PBL mice resulted in reduced levels of surface CD4 expression on T cells. Mice infected with HIV-1 and treated on a weekly basis with scFvCD7-9R-siRNA complexes targeting a combination of viral genes and the host coreceptor molecule CCR5 successfully maintained CD4/CD3 T cell ratios up to 4 weeks after infection in contrast to control mice that displayed a marked reduction in CD4 T cell numbers. p24 antigen levels were undetectable in 3 of the 4 protected mice. scFvCD7-9R/antiviral siRNA treatment also helped maintain CD4 T cell numbers with reduced plasma viral loads in Hu-PBL mice reconstituted with PBMC from donors seropositive for HIV, indicating that this method can contain viral replication even in established HIV infections. Our results show that scFvCD7-9R could be further developed as a potential therapeutic for HIV-1 infection.

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Mutagenesis of Slow Growing Rhizobium japonicum by Transposon Tn5 (Transposon Tn5를 이용한 Slow growing Rhizobium japonicum의 돌연변이 유도)

  • Kim, Sung-Hoon;Rhee, Yoon;Sun, Dae-Kyu;Yoo, Ick-Dong
    • Korean Journal of Microbiology
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    • v.26 no.4
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    • pp.305-311
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    • 1988
  • The spectinomycin resistant strain of slow growing R. japonicum R-168 was selected to be participated in conjugation with E. coli WA803/pGS9. Tn5 was introduced from suicide vector pGS9 into R. japonicum R-168 $spr^{r}$ chromosome at the frequency of $1.0\times 10^{-5}-5.0\times 10^{-7}$ and the transconjugante were selected on the yeast extract-mannitol plate containing kanamycin ($50{\mu}$g/ml) and spectinomycin ($100{\mu}$g/ml) after 8-9 days incubation. All transconjugants we tested were found to contain Tn 5 DNA on their genome, which was confirmed by Southern hybridization experiments. R. japonicum RNa75, which had been selected through plant test, was found to be defective in symbiotic nitrogen fixing ability and the production of leghemoglobin in soybean nodules formed by the inoculation of this mutant. In addition, this mutant strain hardly developed nitrogenase activity asymbiotically in contrast with the wild type strain, indicating that some nitrogen fixing gene might be blocked in this strain and the production of leghemoglobin could be decreased by the interference in nitrogen fixing genes.

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Saprolegnia parasitica Isolated from Rainbow Trout in Korea: Characterization, Anti-Saprolegnia Activity and Host Pathogen Interaction in Zebrafish Disease Model

  • Shin, Sangyeop;Kulatunga, D.C.M.;Dananjaya, S.H.S.;Nikapitiya, Chamilani;Lee, Jehee;De Zoysa, Mahanama
    • Mycobiology
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    • v.45 no.4
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    • pp.297-311
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    • 2017
  • Saprolegniasis is one of the most devastating oomycete diseases in freshwater fish which is caused by species in the genus Saprolegnia including Saprolegnia parasitica. In this study, we isolated the strain of S. parasitica from diseased rainbow trout in Korea. Morphological and molecular based identification confirmed that isolated oomycete belongs to the member of S. parasitica, supported by its typical features including cotton-like mycelium, zoospores and phylogenetic analysis with internal transcribed spacer region. Pathogenicity of isolated S. parasitica was developed in embryo, juvenile, and adult zebrafish as a disease model. Host-pathogen interaction in adult zebrafish was investigated at transcriptional level. Upon infection with S. parasitica, pathogen/antigen recognition and signaling (TLR2, TLR4b, TLR5b, NOD1, and major histocompatibility complex class I), pro/anti-inflammatory cytokines (interleukin $[IL]-1{\beta}$, tumor necrosis factor ${\alpha}$, IL-6, IL-8, interferon ${\gamma}$, IL-12, and IL-10), matrix metalloproteinase (MMP9 and MMP13), cell surface molecules ($CD8^+$ and $CD4^+$) and antioxidant enzymes (superoxide dismutase, catalase) related genes were differentially modulated at 3- and 12-hr post infection. As an anti-Saprolegnia agent, plant based lawsone was applied to investigate on the susceptibility of S. parasitica showing the minimum inhibitory concentration and percentage inhibition of radial growth as $200{\mu}g/mL$ and 31.8%, respectively. Moreover, natural lawsone changed the membrane permeability of S. parasitica mycelium and caused irreversible damage and disintegration to the cellular membranes of S. parasitica. Transcriptional responses of the genes of S. parasitica mycelium exposed to lawsone were altered, indicating that lawsone could be a potential anti-S. parasitica agent for controlling S. parasitica infection.

Physiological Function of Isoflavones and Their Genetic and Environmental Variations in Soybean (콩 Isoflavone의 생리활성 기능과 함량 변이)

  • Kim Yong-Ho;Kim Seok-Dong;Hong Eun-Hi;Ahn Wan-Sik
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.41 no.spc1
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    • pp.25-45
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    • 1996
  • Soyfoods have potential roles in the prevention and treatment of chronic diseases, most notably cancer, osteoporosis, and heart disease. There is evidence that carcinogenesis are supressed by isolated soybean derived products in vivo such as a protease inhibitor, phytic acid, saponins and isoflavones. It is believed that supplementation of human diets with soybean products markedly reduces human cancer mortality rates. Especially, recent papers recognize the potential benefit of soybean isoflavone components for reducing the risk of various cancers. Isoflavones exhibit a multitude of medicinal effects that influence cell growth and regulation, which may have potential value in the prevention and treatment of cancer. In addition to potential biological effects, soybean isoflavones have the important physiological functions such as the induction of Bradyrizobium japonicum nod genes and the responses of soybean tissues to infection by Phytophthora megasperma as well as biochemical activities such as antifungal and antibacterial actions. Genistin, daidzin, glycitin and their aglycone (genistein, daidzein, glycitein) are the principal isoflavones found in soybean. Malonyl and acetyl forms have also been detected but they are thermally unstable and are usually transformed during the processing in glucoside form. Most soy products, with the exception of soy sauce, alcohol-extracted soy protein concentrate, and soy protein isolate, have total isoflavone concentrations similar to those in the whole soybean. Soybean-containing diets inhibit mammary tumorigenesis in animal models of breast cancer, therefore, it is possible that dietary isoflavones are an important factor accounting for the lower incidence and mortality from breast cancer. Of the total soybean seed isoflavones, $80\~90\%$ were located in cotyledons, with the remainder in the hypocotyls. The hypocotyls had a higher concentrations of isoflavones on a weight basis compared with cotyledons. Isoflavone contents were influenced by genetics, crop years, and growth locations. The effect of crop year had a greater impact on the isoflavone contents than that of location. The climate condition might be the attribution factor to variation in isoflavone contents. Also, while the isoflavone content of cotyledons exhibited large variations in response to high temperature during seed development, hypocotyls showed high concentration in isoflavone content. So, it is concluded that one of the factors affecting isoflavone content in soybean seeds is temperature during seed development. High temperature, especially in maturity stage, causes lower isoflavone content in soybean seed. It is also suggested that there may exist a different mechanism to maintain isoflavone contents between cotyledon and seed hypocotyls. In a conclusion, soy foods may be able to have a significant beneficial impact on public health.

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