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Incheon, South Korea

Seo H.-H.,BIO FD and C Co. | Park S.,Chonnam National University | Park S.,Experiment Research Institute | Oh B.-J.,Biological Control Center | And 4 more authors.
PLoS ONE | Year: 2014

Functional characterization of a defensin, J1-1, was conducted to evaluate its biotechnological potentiality in transgenic pepper plants against the causal agent of anthracnose disease, Colletotrichum gloeosporioides. To determine antifungal activity, J1-1 recombinant protein was generated and tested for the activity against C. gloeosporioides, resulting in 50% inhibition of fungal growth at a protein concentration of 0.1 mg·mL-1. To develop transgenic pepper plants resistant to anthracnose disease, J1-1 cDNA under the control of 35S promoter was introduced into pepper via Agrobacterium-mediated genetic transformation method. Southern and Northern blot analyses confirmed that a single copy of the transgene in selected transgenic plants was normally expressed and also stably transmitted to subsequent generations. The insertion of T-DNA was further analyzed in three independent homozygous lines using inverse PCR, and confirmed the integration of transgene in non-coding region of genomic DNA. Immunoblot results showed that the level of J1-1 proteins, which was not normally accumulated in unripe fruits, accumulated high in transgenic plants but appeared to differ among transgenic lines. Moreover, the expression of jasmonic acid-biosynthetic genes and pathogenesis-related genes were up-regulated in the transgenic lines, which is co-related with the resistance of J1-1 transgenic plants to anthracnose disease. Consequently, the constitutive expression of J1-1 in transgenic pepper plants provided strong resistance to the anthracnose fungus that was associated with highly reduced lesion formation and fungal colonization. These results implied the significance of the antifungal protein, J1-1, as a useful agronomic trait to control fungal disease. © 2014 Seo et al.

Moh S.H.,BIO FD and C Co. | Cho S.H.,Korea University | Kim Y.J.,Chungwoon University | Cho J.C.,Korea University | Lee B.W.,Chungwoon University
Communications in Computer and Information Science | Year: 2012

In this study, sodium hyaluronate was hydrolyzed in order to produce low molecular weight sodium hyaluronate and acetylated low molecular weight sodium. The sodium hyaluronate produced in this study had the average molecular weight that ranged from several hundreds of thousands to 3,500 Da by adjusting the acid concentration and reaction time, and then the sodium hyaluronate was converted to acetylated low molecular weight sodium hyaluronate. The testing on cytotoxicity in HaCaT Human Skin Keratinocytes, all the samples, which were processed with the low molecular weight sodium hyaluronate and acetylated low molecular weight sodium, appeared to have no cytotoxicity. The testing result on anti-wrinkle activity in CCD 986-sk human dermal fibroblasts showed that as the molecular weight low molecular weight sodium hyaluronate reduced, the effects on anti-wrinkle activity was getting better, and increase in the amount of PIP synthesis at the LMHA-5, LMHA-6, which were the acetylated samples, had an excellent effect on anti-wrinkle activity. And the measurement results of anti-inflammation showed that as the molecular weight reduced, the amount of NO generated was getting smaller, and the acetylated samples appeared to have much smaller amount of NO, so that acetylated low molecular weight sodium hyaluronate could be considered to have an excellent anti-inflammation effect. © 2012 Springer-Verlag.

Cho S.H.,Korea University | Moh S.H.,BIO FD and C Co. | Kim Y.J.,Chungwoon University | Cho J.C.,Korea University | Kim Y.S.,Kangwon National University
International Journal of Bio-Science and Bio-Technology | Year: 2014

CM-1,3-β-Glucan was synthesis by introducing carboxy methyl group with molar ratio of monochloroacetic acid, and cytotoxicity, antioxidant, whitening effect, anti-inflammation, and anti-wrinkle characteristics were examined It was confirmed that carboxymethyl was quantitatively introduced to β-glucan. As a result of conducting test on cell stability of human keratinocyte cell lines (HaCaT), it did not present cytoxicity at both 0.1% and 0.5% concentration compared to control group and it was revealed to have no cytotoxicity at all. Presented high cell proliferation rate compared to control group at 0.1% concentration. Although anti-inflammation effect was confirmed with the reduction of COX-2 expression in CMB-1, CMB-5, CMB-6, CMBA-1, and CMBA-3 at 0.1% concentration as a result of conducting COX-2 expression influence test against animal cells, an anti-inflammatory test, it was revealed to have no effect in other substances. As a result of conducting MMP-1 expression influence test, an anti-wrinkle test, great anti-wrinkle effect was confirmed as MMI expression reduced in CMR-1, CMB-2, CMB-3, CMB-4, CMB-6, CMBA-1, and CMBA-3 at 0.1% concentration and CMB-1, CMB-2, CMB-3, CMB-4, and CMBA-1 at 0.5% concentration. © 2014 SERSC.

Nguyen M.T.,University of Ulsan | Koo B.-K.,University of Ulsan | Thi Vu T.T.,University of Ulsan | Song J.-A.,University of Ulsan | And 5 more authors.
PLoS ONE | Year: 2014

Human growth hormone (hGH) is synthesized by somatotroph cells of the anterior pituitary gland and induces cell proliferation and growth. This protein has been approved for the treatment of various conditions, including hGH deficiency, chronic renal failure, and Turner syndrome. Efficient production of hGH in Escherichia coli (E. coli) has proven difficult because the E. coli-expressed hormone tends to aggregate and form inclusion bodies, resulting in poor solubility. In this study, seven N-terminal fusion partners, hexahistidine (His6), thioredoxin (Trx), glutathione S-transferase (GST), maltosebinding protein (MBP), N-utilization substance protein A (NusA), protein disulfide bond isomerase (PDI), and the b′a′ domain of PDI (PDIb′a′), were tested for soluble overexpression of codon-optimized hGH in E. coli. We found that MBP and hPDI tags significantly increased the solubility of the hormone. In addition, lowering the expression temperature to 18°C also dramatically increased the solubility of all the fusion proteins. We purified hGH from MBP-, PDIb′a′-, or Trx-tagged hGH expressed at 18°C in E. coli using simple chromatographic techniques and compared the final purity, yield, and activity of hGH to assess the impact of each partner protein. Purified hGH was highly pure on silver-stained gel and contained very low levels of endotoxin. On average, ∼37 mg, ∼12 mg, and ∼7 mg of hGH were obtained from 500 mL-cell cultures of Trx-hGH, MBP-hGH, and PDIb′a′-hGH, respectively. Subsequently, hGH was analyzed using mass spectroscopy to confirm the presence of two intra-molecular disulfide bonds. The bioactivity of purified hGHs was demonstrated using Nb2-11 cell. © 2014 Nguyen et al.

Lee G.,Korea Advanced Institute of Science and Technology | Park S.Y.,Korea University | Yum S.,Korea Advanced Institute of Science and Technology | Woo S.,Korea Advanced Institute of Science and Technology | And 6 more authors.
Biochip Journal | Year: 2012

Countless species occur in the marine microalgal domain. Some are used as health functional foods or medical products but many species are harmful such as those that cause the red tide. Therefore, it is necessary to conduct prompt and accurate identification of microalgal species. As it is quite difficult to accurately distinguish all species in terms of morphology, we performed DNA barcoding analysis using molecular markers for more accurate and rapid screening. DNA barcoding analysis, i. e., DNA chip technology, is a powerful method for studies on microalgal taxonomy and biodiversity. We used the mitochondrial cytochrome c oxidase subunit I (mtCOI) as a barcoding gene to identify microalgal species. In this study, the diversity and phylogenetic differences among different microalgae were analyzed. Additionally, a microalgal species-specific probe was screened by 21-23 bp and the result was printed on silylated slide for use in a robotic microarrayer. As a result, we performed a DNA chip assay for each of 25 microalgal species and determined that the COI barcode gene was suitable as a marker gene, as it could identify various microalgae from the Korean South Sea by species. © 2012 The Korean BioChip Society and Springer-Verlag Berlin Heidelberg.

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