Tae-Geum Kim

Chonbuk National University, Tsiuentcheou, North Jeolla, South Korea

Are you Tae-Geum Kim?

Claim your profile

Publications (64)108.82 Total impact

  • Mi-Young Kim · Yong-Suk Jang · Moon-Sik Yang · Tae-Geum Kim
    [Show abstract] [Hide abstract]
    ABSTRACT: Although plant-based vaccines have many advantages, their use is limited by low expression of antigen genes in transgenic plants, which results in low immune responses and immune tolerance. To overcome this problem, Nicotiana benthamiana was used to produce the consensus domain III of dengue virus envelope glycoprotein (E) via agroinfiltration with a plant virus-based expression system. The binding of E glycoprotein to a receptor is important for dengue virus entry into host cells and results in human disease. Consensus domain III of dengue virus E glycoprotein (cEDIII) is immunogenic and can elicit neutralizing antibodies against all four serotypes of dengue virus. A DNA fragment encoding cEDIII and M cell-targeting ligand fused to cEDIII (cEDIII-Co1) were constructed in a viral vector and introduced into tobacco plant cells by Agrobacterium-mediated infiltration. The cEDIII and cEDIII-Co1 fusion proteins were detected in protein extracts from agroinfiltrated leaves by Western blot analysis. The plant-produced cEDIII and cEDIII-Co1 fusion proteins composed 5.2 and 4.8 mg/g of dry weight of leaf tissues, respectively. These results suggest that the high expression of dengue virus cEDIII in plants with a plant virus-based expression system can overcome the low expression level to improve the feasibility of plant-based vaccines.
    No preview · Article · May 2015 · Plant Cell Tissue and Organ Culture
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Vascular endothelial growth factors (VEGFs) are secreted by tumor cells and other cells exposed to hypoxia, and play a critical role in the development and differentiation of the vascular system. In this study, we investigated the production of functional recombinant human VEGF165 (rhVEGF165) in transgenic rice cell suspension culture. Complementary DNA was synthesized from human leukemia HL60 cells and cloned into expression vectors under the control of the rice α-amylase 3D (RAmy3D) promoter. The rice seed (Oryza sativa L. cv. Dongjin) was transformed with this recombinant vector by the Agrobacterium mediated method and the integration of the target gene into the plant genome was confirmed by genomic PCR. The expression of rhVEGF165 in the rice cells was determined by Northern blot and Western blot analyses. The accumulated rhVEGF165 protein in the culture medium was 19 mg/L after 18days of culturing in a sugar-free medium. The rhVEGF165 was purified using aheparin HP column and its biological activity was tested on human umbilical vein endothelial cells (HUVECs). The purified rhVEGF165 significantly increased the proliferative activity of the HUVECs. Therefore, it was demonstrated that functional rhVEGF165 could be produced using transgenic rice suspension culture vector under the control of the RAmy3D promoter.
    Full-text · Article · Sep 2014 · Enzyme and Microbial Technology
  • [Show abstract] [Hide abstract]
    ABSTRACT: Major fimbriae (FimA) of Porphyromonas gingivalis play an important role in the adherence of the bacterium to oral surfaces, making them a target for the development of a vaccine against periodontal disease. In our previous study, monoclonal antibodies to FimA were expressed in tobacco plants using the agroinfiltration method. In this report, for easy purification, monoclonal antibodies to FimA were produced and accumulated in rice callus suspension culture. The accumulated antibodies were purified by protein G-affinity chromatography. The plant-produced monoclonal antibodies inhibited the binding of P. gingivalis to saliva-coated hydroxyapatite beads, as well as the invasion of oral epithelial KB cells due to the bacterium. The antibodies enhanced the intracellular killing of P. gingivalis by polymorphonuclear neutrophils. These results suggest that FimA-specific monoclonal antibodies produced in a rice suspension culture were easily purified and biologically active against P. gingivalis, and thus may be used for passive immunization to prevent P. gingivalis-induced periodontal disease.
    No preview · Article · Aug 2014 · Plant Cell Tissue and Organ Culture
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Dengue is a disease caused by dengue virus and represents the most important arthropod-borne viral disease in humans. Dengue virus enters host cells via binding of envelope glycoprotein (E) to a receptor. In this study, plant expression vectors containing native and synthetic glycoprotein E genes (sE) modified based on plant-optimized codon usage and fused with an ER retention signal were constructed under control of the rice amylase 3D promoter expression system. Plant expression vectors were introduced into rice callus (Oryza sativa L. cv. Dongin) via particle bombardment-mediated transformation. The integration and expression of target genes were confirmed in the transgenic callus by genomic DNA PCR and Northern blot analyses, respectively. The plant-codon optimized sE gene with an ER retention signal showed high protein production levels based on Western blot analysis of approximately 18.5 ug/g dried calli weight by immunoblot-based densitometric analysis. These results suggest that the plant-codon optimized sE gene with an ER retention signal was highly produced in the transgenic rice callus.
    Full-text · Article · Jul 2014 · Molecular Biotechnology
  • [Show abstract] [Hide abstract]
    ABSTRACT: We report a feasibility study for expressing the LTB protein (Escherichia coli heat-labile enterotoxin B subunit) via Agrobacterium-mediated transformation of tomato (Solanum lycopersicum L.). We produced five regenerated plants obtained on the selection medium supplemented with an antibiotic. Stable integrations of the LTB gene into the genome of these plants were confirmed by Southern blot hybridization. Western blot analysis showed that only two of the five T0 transgenic tomato plants expressed the pentameric LTB protein in the fruits. An enzyme-linked immunosorbent assay indicated that these two plants synthesized the LTB protein bound specifically to GM1 ganglioside, suggesting that the LTB subunits formed active pentamers. The LTB protein produced in tomatoes can be a potential candidate for inexpensive, safe, and effective plant-based vaccines.
    No preview · Article · Jan 2014 · Czech Journal of Genetics and Plant Breeding
  • [Show abstract] [Hide abstract]
    ABSTRACT: A rice cell suspension culture with the rice α-amylase 3D promoter expression system which is induced by sucrose starvation was previously reported to generate a good yield of recombinant proteins. However, this expression system is limited by the accumulation of undesirable α-amylase and proteases in the culture medium. Rice α-amylase is a dominant protein at 43 % of total secreted proteins, and cysteine proteinase (CysP) is a major secreted protease in rice cell suspension cultures following induction via sugar depletion. Here, we nearly eliminated rice α-amylase and CysP proteinase via RNA interference (RNAi) technology to improve the recombinant protein yield in rice cell suspension culture. The effects of RNAi were characterized by sodium dodecyl sulfate–polyacrylamide gel electrophoresis, Western blot analysis with anti-CysP antibody, and quantitative real-time reverse transcription polymerase chain reaction analysis (RT-PCR). The mRNA levels of α-amylase and CysP were reduced by 94.8 and 95.0 %, respectively. Transgenic rice cell suspension cultures expressing both human granulocyte–macrophage colony-stimulating factor (hGM-CSF) and ihpRNA of α-amylase and CysP genes evidenced a reduction of α-amylase and CysP activity and up to 2.4-fold improvement of hGM-CSF production compared to that in a transgenic cell line expressing hGM-CSF only. Our rice cell suspension culture for the reduction of α-amylase accumulation and protease activity in the culture medium could improve recombinant protein production as an efficient protein expression system using RNA interference technology in plant biotechnology.
    No preview · Article · Jul 2013 · Plant Cell Tissue and Organ Culture
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: In this work, the characterizations of the LTB (Escherichia coli heat-labile toxin B subunit) transgenic watercresses through Agrobacterium tumefaciens-mediated transformation (A1 and A3-A5) and by using biolistic method (B1 and B4) were investigated. Generally, their growth is not remarkably different from the wild-type. Their physiological and biochemical characteristics are relatively different in which plant A1 has highest values such as pigment (0.92 mg g-1), photosynthetic rate (23.39 mgCO2 [dm2]-1 h-1), dry matter (7.42%), vitamin C (0.34 mg g-1), calcium (0.83 mg g-1), and potassium (2.47 mg g-1). The dry matter and calcium of all the transgenic watercresses and the wild-type are the same content. Southern blot hybridization showed the transgenic plants contain 1-2 copies in the genome. LTB protein strongly expresses in all the transgenic plants with contents from 1.16 to 1.46% of total soluble protein. The GM1-ELISA binding assay indicated that plant-derived LTB protein bound to GM1-ganglioside receptors.
    Full-text · Article · Apr 2013 · Journal of Plant Biochemistry and Biotechnology
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: The spread of dengue (DEN) virus is becoming a major concern due to the possibility of primary infection with one of the four dengue serotypes (DEN 1-4) and secondary infection with other heterotypes, which can further aggravate clinical manifestations. A gene encoding consensus envelope protein domain III (cEDIII) of dengue virus with neutralizing activity against four dengue virus serotypes was fused to M cell-targeting peptide ligand (Co1) to increase its mucosal immunogenicity and was introduced into rice calli under the control of the inducible rice amylase 3D promoter expression system. The integration and expression of scEDIII-Co1 fusion gene in transgenic rice calli were confirmed by genomic DNA PCR amplification, Northern and Western blot analyses, respectively. The deliveries of cEDIII-Co1 fusion proteins into mucosal immune inductive site (including M cells) were confirmed by in vitro and in vivo antigen uptake assays. These results showed that plant-produced M cell-targeting peptide ligand, Co1, fusion antigen proteins have the potential to be targeted to the mucosal immune system for improvement of immune responses.
    Full-text · Article · Dec 2012 · Molecular Biotechnology
  • Nguyen-Xuan Huy · Sae-Hae Kim · Moon-Sik Yang · Tae-Geum Kim
    [Show abstract] [Hide abstract]
    ABSTRACT: To increase immune responses of plant-based vaccines in intestine mucosal immune systems, a synthetic neutralizing epitope (sCOE) gene of porcine epidemic diarrhea virus (PEDV) was fused with M cell-targeting ligand (Co1) and introduced into a plant expression vector under the control of rice amylase 3D promoter. The sCOE–Co1 fusion gene was introduced into rice calli via the particle bombardment-mediated transformation method. The stable integration and transcriptional expression of the sCOE–Co1 fusion gene was confirmed by genomic DNA PCR amplification and Northern blot analysis, respectively. The expression of the COE–Co1 fusion protein was confirmed by immunoblot analysis. The highest expression level of the COE–Co1 fusion protein reached 0.083 % of the total soluble protein according to quantitative densitometry of Western blot analysis. Mice immunized with transgenic rice calli protein extracts induced significant serum IgG and fecal IgA antibody levels against purified bacterial COE. The systemic and mucosal immune responses were confirmed by measuring COE-specific IgG and IgA antibody-secreting cells in the lymphocytes extracted from the spleen and COE-specific IgA antibody-secreting cells in the Peyer’s patches from immunized mice. These results indicated that oral immunization of plant-produced COE–Co1 fusion protein could elicit efficient systemic and mucosal immune responses against the COE antigen. Key message Neutralizing epitope from porcine epidemic diarrhea virus-M cell targeting ligand fusion protein was produced in transgenic rice calli and elicited systemic and mucosal immune responses by oral administration in mice.
    No preview · Article · Jun 2012 · Plant Cell Reports
  • Source
    Mi-Young Kim · Moon-Sik Yang · Tae-Geum Kim
    [Show abstract] [Hide abstract]
    ABSTRACT: An effective dengue vaccine should elicit immune responses against all four different dengue virus serotypes. This study optimized the codon usage of a gene encoding consensus dengue virus envelope protein domain III (cEDIII) with cross-neutralizing activity against four dengue virus serotypes for plant expression. Then, a plant expression vector was constructed with this gene under the control of the rice amylase 3D promoter (RAmy3D), which is a strong inducible promoter under sugar starvation conditions. The synthetic cEDIII gene was fused with the RAmy3D signal peptide and ER retention signal, SEKDEL, and was introduced into rice callus by particle bombardment-mediated transformation. The integration and expression of cEDIII gene in transgenic rice callus was confirmed by genomic DNA PCR amplification, Northern blot analysis, and western blot analysis, respectively. Densitometric analysis determined that the highest expression level of the cEDIII protein in lyophilized rice callus was approximately 0.45 mg g−1. These results suggest that it is feasible to use transgenic rice callus to produce the consensus dengue virus envelop protein domain III for edible vaccine purposes.
    Full-text · Article · Jun 2012 · Plant Cell Tissue and Organ Culture
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: In this study, three LTB (Escherichia coli heat-labile toxin B subunit) transgenic tomato lines (#1-3) were grown in the pot to characterize their growth and development, and evaluate fruit and LTB subunit productivity. Generally, the growth and development of transgenic tomato lines are not significantly different with non-transgenic tomato cultivar, “311” (commercial cultivar as reference). All of them took 120 days to final harvest, and number of fruit is near equivalent. However, fruit quality characteristics are relatively different and line #3 has highest values of dry matter (6.02%), reducing sugar (2.51%) and degree Brix (6.40%). The vitamin C and acidity of all transgenic lines and the control are the same content. LTB subunit only expressed in two lines #1 and #3 with contents of 1.04% and 1.19% of total soluble protein, respectively.
    Full-text · Article · May 2012
  • [Show abstract] [Hide abstract]
    ABSTRACT: Porphyromonas gingivalis, a gram-negative anaerobic oral bacterium, causes periodontal disease by binding to saliva-coated oral surfaces. The FimA protein from P. gingivalis is a crucial pathogenic component of the bacterium and a target for vaccine development against periodontal disease. Complementary DNAs encoding the heavy and light chains of two monoclonal antibodies that bind specifically to the FimA protein were cloned into a plant expression vector under the control of the duplicated Cauliflower Mosaic Virus 35S promoter, and agroinfiltration was used to allow the vectors to infiltrate tobacco plants. The expressions of the heavy and light chains in the leaf tissue were detected using antibodies specific to each antibody chain. Western blot analysis showed the specific binding of the plant-derived monoclonal antibodies to the native FimA protein purified from P. gingivalis. Our finding that plant-derived monoclonal antibodies bound specifically to the native FimA protein indicates that plantderived monoclonal antibodies can protect against P. gingivalis invasion.
    No preview · Article · Apr 2012 · Biotechnology and Bioprocess Engineering
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Dengue (DEN) is one of the most important emerging mosquito-borne viral human diseases. Therefore, an effective dengue vaccine with immune responses against all four dengue virus serotypes is highly needed. A fusion gene encoding a synthetic consensus envelope protein domain III (scEDIII) of dengue virus with neutralizing activity against the four dengue virus serotypes and with the B subunit of cholera toxin (CTB) to increase its mucosal immunogenicity was constructed and was introduced into rice callus under the control of the inducible rice amylase 3D promoter expression system. The integration and expression of the CTB-scEDIII fusion gene in transgenic rice callus were confirmed by genomic DNA PCR amplification, Northern, and Western blot analyses, respectively. The biological binding activity of the CTB-cEDIII fusion protein to its GM1-ganglioside receptor was confirmed via GM1-ELISA with anti-CT and anti-dengue virus antibodies. Delivery of the CTB-cEDIII fusion protein into mucosal immune inductive sites (including M cells) in BALB/c mice was confirmed by in vitro and in vivo antigen uptake assays. These results showed that the CTB-cEDIII fusion protein was produced in the transgenic rice callus, and that plant-produced ligand fusion antigen proteins have the potential to be targeted to the mucosal immune system for improvement of the overall immune responses.
    Full-text · Article · Mar 2012 · Plant Cell Tissue and Organ Culture
  • [Show abstract] [Hide abstract]
    ABSTRACT: The cholera toxin B subunit (CTB), a nontoxic molecule with potent biological properties, is a powerful mucosal and parenteral adjuvant that induces a strong immune response against co-administered or coupled anti- gens. A gene encoding CTB, which was modified based on the optimized codon usage in the plant, was synthesized and fused to the endoplasmic reticulum retention signal KDEL to enhance its expression level in plants. The syn- thetic CTB (sCTB) gene was introduced into a plant ex- pression vector adjacent to the CaMV 35S promoter, and was transformed into tomato using an Agrobacterium- mediated transformation method. The integration of the sCTB gene into the genomic DNA of transgenic plants was confirmed by genomic DNA PCR amplification. The syn- thesis and assembly of CTB protein in transgenic plants was demonstrated through immunoblot analysis and G - ELISA. The highest amount of CTB protein produced in transgenic tomatoes was approximately 0.9% of total soluble fruit protein which was 10-fold greater than the previously 0.081%. G -ELISA indicated that plant-synthesized CTB protein bound specifically to G -gangliosides, suggesting that the CTB subunits formed active pentamers.
    No preview · Article · Jun 2011 · Biotechnology and Bioprocess Engineering
  • [Show abstract] [Hide abstract]
    ABSTRACT: A gene encoding the B subunit of the enterotoxigenic Escherichia coli heat-labile enterotoxin (LTB) was adapted to the optimized plant coding sequence, and fused to the endoplasmic reticulum retention signal SEKDEL in order to enhance its expression level and protein assembly in plants. The synthetic LTB (sLTB) gene was placed into a plant expression vector under the control of the CaMV 35S promoter, and subsequently introduced into the watercress (Nasturtium officinale L.) plant by the Agrobacterium-mediated transformation method. The integration of the sLTB gene into the genomic DNA of transgenic plants was confirmed by genomic DNA PCR amplification. The assembly of plant-produced LTB protein was detected by western blot analysis. The highest amount of LTB protein produced in transgenic watercress leaf tissue was approximately 1.3% of the total soluble plant protein. GM1-ganglioside enzyme-linked immunosorbent assay indicated that plant-synthesized LTB protein bound specifically to GM1-ganglioside, which is the receptor for biologically active LTB on the cell surface, suggesting that the plant-synthesized LTB subunits formed biologically active pentamers.
    No preview · Article · Apr 2011 · Plant Cell Tissue and Organ Culture
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: FimA of Porphyromonas gingivalis, a major pathogen in periodontitis, is known to be closely related to the virulence of these bacteria and has been suggested as a candidate for development of a vaccine against periodontal disease. In order to develop a passive immunization method for inhibiting the establishment of periodontal disease, B hybridoma clones 123-123-10 and 256-265-9, which produce monoclonal antibodies (Mabs) specific to purified fimbriae, were established. Both mAbs reacted with the conformational epitopes displayed by partially dissociated oligomers of FimA, but not with the 43 kDa FimA monomer. Gene sequence analyses of full-length cDNAs encoding heavy and light chain immunoglobulins enabled classification of the genes of mAb 123-123-10 as members of the mVh II (A) and mVκ I subgroups, and those of mAb 256-265-9 as members of the mVh III (D) and mVκ I subgroups. More importantly, 50 ng/mL of antibodies purified from the culture supernatant of antibody gene-transfected CHO cells inhibited, by approximately 50%, binding of P. gingivalis to saliva-coated hydroxyapatite bead surfaces. It is expected that these mAbs could be used as a basis for passive immunization against P. gingivalis-mediated periodontitis.
    Preview · Article · Mar 2011 · Microbiology and Immunology
  • [Show abstract] [Hide abstract]
    ABSTRACT: The cholera toxin B subunit (CTB), a nontoxic molecule with potent biological properties, is a powerful mucosal and parenteral adjuvant that induces a strong immune response against co-administered or coupled antigens. A gene encoding CTB, which was modified based on the optimized codon usage in the plant, was synthesized and fused to the endoplasmic reticulum retention signal KDEL to enhance its expression level in plants. The synthetic CTB (sCTB) gene was introduced into a plant expression vector adjacent to the CaMV 35S promoter, and was transformed into tomato using an Agrobacterium-mediated transformation method. The integration of the sCTB gene into the genomic DNA of transgenic plants was confirmed by genomic DNA PCR amplification. The synthesis and assembly of CTB protein in transgenic plants was demonstrated through immunoblot analysis and GM1-ELISA. The highest amount of CTB protein produced in transgenic tomatoes was approximately 0.9% of total soluble fruit protein which was 10-fold greater than the previously 0.081%. GM1-ELISA indicated that plant-synthesized CTB protein bound specifically to GM1-gangliosides, suggesting that the CTB subunits formed active pentamers.
    No preview · Article · Jan 2011
  • Mi-Young Kim · Moon-Sik Yang · Tae-Geum Kim
    [Show abstract] [Hide abstract]
    ABSTRACT: Dengue virus enters into host cells by binding envelope glycoprotein (E) to a receptor and causes human disease. Domain III of dengue virus E glycoprotein (EIII) is immunogenic and capable of inducing neutralizing antibodies. A DNA fragment encoding EIII (amino acids 297–394) was introduced into tobacco plant cells (Nicotiana tabacum L. cv MD609) by Agrobacterium tumefaciens-mediated transformation methods. The integration of EIII gene was observed in the genomic DNA of transgenic plants by PCR DNA amplification methods. The transcripts of EIII gene were detected in the transgenic plant leaf tissues by Northern blot analysis. The EIII protein was detected in transgenic plant leaf extracts by Western blot analysis. Based on the results of the Western blot analysis, the expression level of plant-produced EIII protein was between 0.13 and 0.25% of the total soluble protein in transgenic plant leaf tissues. The production of domain III of dengue virus E glycoprotein (EIII) in transgenic plants demonstrates the feasibility of using plant-based vaccines to prevent infection by the dengue virus.
    No preview · Article · Dec 2010 · Biotechnology and Bioprocess Engineering
  • [Show abstract] [Hide abstract]
    ABSTRACT: Plant-based vaccines have been produced in transgenic plants including tobacco, potatoes, corn, and rice. However, these plants are not suitable for administration without cooking. To overcome this obstacle, a fusion gene encoding the synthetic enterotoxigenic Escherichia coli heat-labile enterotoxin B subunit genetically fused with a synthetic neutralizing epitope of porcine epidemic diarrhea virus (sLTB-sCOE) was introduced into lettuce cells (Lactuca sativa) by Agrobacterium-mediated transformation methods. The integration and expression of the sLTB-sCOE fusion gene was confirmed in transgenic lettuce by genomic DNA PCR amplification and Northern blot analysis, respectively. Synthesis and assembly of the LTB-COE fusion protein into oligomeric structures with pentamer size were observed in transgenic plant extracts by Western blot analysis with anti-LTB or anti-COE antibodies. The binding of plantproduced LTB-COE to intestinal epithelial cell membrane glycolipid receptors was confirmed by GM1-ganglioside enzyme-linked immunosorbent assay (GM1-ELISA). Based on the ELISA results, LTB-COE fusion protein made up about 0.026∼0.048% of the total soluble protein in the transgenic lettuce leaf tissues. The synthesis and assembly of LTB-COE monomers into biologically active oligomers in transgenic lettuce leaf tissues demonstrates the feasibility of using uncooked edible plant-based vaccines for mucosal immunization.
    No preview · Article · Dec 2010 · Biotechnology and Bioprocess Engineering
  • Nguyen-Xuan Huy · Moon-Sik Yang · Tae-Geum Kim
    [Show abstract] [Hide abstract]
    ABSTRACT: Transgenic plants have been used as a safe and economic expression system for the production of edible vaccines. A synthetic cholera toxin B subunit gene (CTB) was fused with a synthetic neutralizing epitope gene of the porcine epidemic diarrhea virus (sCTB-sCOE), and the sCTB-sCOE fusion gene was introduced into a plant expression vector under the control of the ubiquitin promoter. This plant expression vector was transformed into lettuce (Lactuca sativa L.) using the Agrobacterium-mediated transformation method. Stable integration and transcriptional expression of the sCTB-sCOE fusion gene was confirmed using genomic DNA PCR analysis and northern blot analysis, respectively. The results of western blot analysis with anti-cholera toxin and anti-COE antibody showed the synthesis and assembly of CTB-COE fusion protein into oligomeric structures with pentameric sizing. The biological activity of CTB-COE fusion protein to its receptor, G(M1)-ganglioside, in transgenic plants was confirmed via G(M1)-ELISA with anti-cholera toxin and anti-COE antibody. Based on G(M1)-ELISA, the expression level of CTB-COE fusion proteins reached 0.0065% of the total soluble protein in transgenic lettuce leaf tissues. Transgenic lettuce successfully expressing CTB-COE fusion protein will be tested to induce efficient immune responses against porcine epidemic diarrhea virus infection by administration with raw material.
    No preview · Article · Dec 2010 · Molecular Biotechnology

Publication Stats

716 Citations
108.82 Total Impact Points

Institutions

  • 2001-2014
    • Chonbuk National University
      • • Department of Molecular Biology
      • • Department of Food and Nutrition
      Tsiuentcheou, North Jeolla, South Korea
  • 2009
    • Hue University
      Huế, Thừa Thiên-Huế, Vietnam
  • 2004-2006
    • Loma Linda University
      • • Department of Medicine
      • • Division of Biochemistry
      Loma Linda, California, United States