• Plant Biotechnology Reports
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• v.2 no.1
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• pp.47-58
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• 2008

To select agronomically useful transgenic plants, a large number of transgenic events are initially produced, gene transfer confirmed, and advanced to obtain homozygous lines for testing in field trials. Direct in planta assays for identifying the transgene carriers in the segregating progeny are based on the activity of selectable marker gene and are easy, simple and inexpensive. For this purpose, expression of bar gene as measured by tolerance to damage by glufosinate ammonium, the active ingredient in the herbicide BASTA, was investigated. Dose damage curves were generated by leaf paint tests with BASTA on four genotypes of sorghum. Transgenic plants were characterized in terms of sensitivity to the concentration of glufosinate ammonium. In transgenics, symptoms of BASTA swab tests at different growth stages and PCR analysis for cry1B were carried out and correlated. Germination tests could not be employed for large scale evaluation of transgenic progeny because of mortality of tolerant seedlings after transplantation to soil. Based on the above findings, a simple, inexpensive, time-saving, two-step scheme for effective evaluation of transgenics and their progeny containing bar gene as selection marker using BASTA swab tests is described.

• Korean Journal of Chemical Engineering
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• v.35 no.11
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• pp.2313-2320
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• 2018

Tetramethylene-1,4-bis(N,N-dodecylammonium bromide), cationic gemini surfactant, (12-4-12) was first synthesized with an one-step and shortened procedure and its interfacial and antimicrobial properties were compared with a conventional single-chain cationic surfactant, cetyltrimethylammonium bromide (CTAB). The interfacial and thermodynamic properties of both surfactants reveal that critical micelle concentration (CMC) of this novel synthetic cationic dimeric surfactant is lower than that of cationic monomeric surfactant at almost 15 times of its magnitude, which is due to the increase in hydrophobicity of the surfactant molecules by having dual hydrocarbon chains. In comparison with CTAB, the produced compound 12-4-12 yields much better interfacial and thermodynamic properties. The antimicrobial activities of the synthesized gemini surfactant were tested against eight strains of bacteria, as well as two strains of fungi. The results showed that both 12-4-12 compound and CTAB exhibited higher inhibitory effects on the growth of Gram-positive bacteria and fungi than that of Gram-negative bacteria. The minimum inhibitory concentrations in molar of 12-4-12 against all tested Gram-negative bacteria were lower than those of CTAB, which is hypothetically due to the lower HLB together with smaller CMC values of our gemini surfactant.

• Mycobiology
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• v.50 no.5
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• pp.345-356
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• 2022

The fungal distribution, diversity, and load were analyzed in the geographically segregated island groundwater systems in Korea. A total of 79 fungal isolates were secured from seven islands and identified based on the internal transcribed spacer (ITS) sequences. They belonged to three phyla (Ascomycota, Basidiomycota, and Chlorophyta), five classes, sixteen orders, twenty-two families, and thirty-one genera. The dominant phylum was Ascomycota (91.1%), with most fungi belonging to the Cladosporium (21.5%), Aspergillus (15.2%), and Stachybotrys (8.9%) genera. Cladosporium showed higher dominance and diversity, being widely distributed throughout the geographically segregated groundwater systems. Based on the diversity indices, the genera richness (4.821) and diversity (2.550) were the highest in the groundwater system of the largest scale. As turbidity (0.064-0.462) increased, the overall fungal count increased and the residual chlorine (0.089-0.308) had low relevance compared with the total count and fungal diversity. Cladosporium showed normal mycelial growth in de-chlorinated sterilized samples. Overall, if turbidity increases under higher fungal diversity, bio-deterioration in groundwater-supplying facilities and public health problems could be intensified, regardless of chlorine treatment. In addition to fungal indicators and analyzing methods, physical hydrostatic treatment is necessary for monitoring and controlling fungal contamination.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.21 no.9
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• pp.559-568
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• 2020

Epidermal growth factor (EGF) is a hormone protein that affects cell growth and proliferation, and has various medical applications. In the present study, the gene of human EGF was codon-optimized with E. coli and the expression vector was constructed by cloning into pRSET. In order to obtain the recombinant human EGF in an active form rather than an inclusion body, chaperone co-expression was attempted along with codon optimization, for the first time. The expressed human EGF was isolated in the pure form by performing Ion Exchange Chromatography in two consecutive runs. ELISA analysis showed that the activity of purified EGF was greater than 99%, which is similar to commercially available EGF. Cell proliferation test confirmed that the recombinant human EGF has the ability to promote cell proliferation of human skin fibroblasts. The human EGF expression system of this study gives a significant amount of protein, and does not require the renaturation step and the additional chromatographic system to remove a fusion contaminant, thereby providing a very useful alternative to conventional expression systems for the preparation of recombinant human EGF.

• Journal of Life Science
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• v.17 no.10
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• pp.1321-1329
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• 2007

Deer antler tissue contains the most rapidly growing bone in the animal kingdom. Thus, it is likely that growing antler tissue is a rich source of local paracrine bone-stimulating factors. Growth factors, at least the insulin-like growth factor (IGF), control the bone-remodelling process. In this study, we tried to isolate and purify IGF-I from fresh antler tissue by the routine isolation and purification of protein. The purification involved ammonium sulfate precipitation, DEAE-Sepharose CL-60 ion-exchange chromatography, CM-Sepharose CL-6B ion-exchange chromatography, and Sephadex G-50 chromatography. Purified fractions from each step were analyzed by high-performance liquid chromatography (HPLC), SDS polyacrylamide gel electrophoresis (SDS-PACE), Dot-blot, and Western-blot methods. Furthermore, the quantification of partially purified IGF-I was calculated by enzyme-linked immunosorbent assays (ELISA) using antibody to human recombinant IGF-1. SDS-PAGE analysis of the final fraction yielded two molecular bands and the signal band was at 12 kDa on the Western-blot film. This purified IGF-I fraction showed a peak at retention time of eight min. The quantity of IGF-I in 20 g deer antler tissue as starting weight was calculated using a standard curve to be 2910 ng/ml, and total IGF-I amount is 0.291 g. The results show that IGF-I, which can be found in deer antler, can be partially purified and quantified by classic protein isolation methods.

• Journal of Korean Society of Forest Science
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• v.101 no.1
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• pp.158-162
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• 2012

The acclimatization of in vitro propagated plants is an important step to produce vigorous plants for clonal forestry and in vitro micro-environment may affect the growth in ex vitro condition. To monitor in vitro environmental effects on the growth in ex vitro condition, several culture systems such semi-solid medium(SS), temporary immersion bioreactor(TIB) and continuous immersion bioreactor(CIB) culture types were tested to compare for the growth of acclimated plants of Liriodendron tulipifera. Results suggested that morphological characters, stomatal conductance, evapotranspiration and chlorophyll contents of acclimated plants were affected by the different of in vitro culture conditions. CIB type of culture was resulted to the lowest value in the biomass of acclimated plants. Net photosynthsis rate of CIB was the same level as those of SS and TIB. However, stomatal conductance, evapotranspiration and $CO_2$ partial pressure in the intercellular air space were lower than those of SS and TIB. The amounts of chlorophyll a, b and carotenoids were also lower than those of the other two culture systems. TIB, showing a little lower or higher value than SS in many growth character, is recommended rather than CIB to produce healthy yellow poplar plants in ex situ condition.

• Korean Chemical Engineering Research
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• v.47 no.5
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• pp.624-629
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• 2009

It is the most important step to screen soluble and insoluble proteins when we attempt to improve the solubility of recombinant proteins through directed evolution approach. Here we show that the solubility of a recombinant protein in vivo can be examined by expressing the recombinant protein with beta-lactamase as a fusion partner. First we constructed an expression system which can produc a fusion protein with the C-terminal of beta-lactamase. Two soluble proteins, i.e. adenine deaminase and aspartate aminotransferase, and insoluble GlcNAc-2-epimerase were cloned into the developed expression vector, respectively. We investigated the effect of the expression of the three recombinant fusion proteins on the growth of E. coli, and confirmed that the solubilities of the recombinant proteins correlated with cell growth rates.

• 한국균학회소식:학술대회논문집
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• 2014.05a
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• pp.21-21
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• 2014

A edible mushroom, Hypsizygus marmoreus is commercially cultivated in Northeast Asia. Japan's annual production is 110,000ton or more. Since 2002, cultivation is expanded in Korea. To investigate the morphological, cultural and microscopic characteristics of Hypsizygus marmoreus, 109 isolates were collected from Korea and other countries. Clamp connection, chlamydospore and arthrospore were present in all tested isolates of H. marmoreus except HYM-002 and HYM-004. Also pilealtrama, gilltrama, basidia, basidiospore and cystidia of fruiting body were no difference among the isolates in the present investigation. Morphological characteristics of fruiting body was that color of pileus was brown and white, irregular as marble, the average size 12~22mm and stipes was $46{\sim}91{\times}6{\sim}10mm$. Isolates HYM-031, HYM-047 and HYM-109 formed grayish-brown pileus with a faint pattern. Molecular analysis with RAPD and ITS rDNA sequence analysis were also performed to check the genetic relationships among H. marmoreus isolates. Based on the RAPD analysis using the URP-PCR, all isolates of H. marmoreus were clustered into large 3 groups but more than 90% showed high similarity. In addition, morphological and geographical differences have been classified as an independent cluster. The brown and white strains enclosed in same cluster. So genetically no significance difference was observed between these two strains. ITS gene sequences of 16 selected isolates which were 640 bp long, were aligned and compared. The similarity in ITS sequence was 94.8 to 99.1% among tested isolates and the H. marmoreus isolates in GeneBank. In conclusion the tested isolates were H. marmoreus. Morphological and molecular observations proved that all tested isolates were belonging to H. marmoreus. For the stable artificial cultivation, composition of optimum media, mature period and light condition were established. Optimal formula of artificial cultivation medium was Douglas sawdust: corn cob: soybean meal: wheat bran = 40:30:15:15. In addition, 7% rice bran and 3% yellow sucrose was the most effective composition for spawn's liquid medium. For the maturation of the isolates was favorable for growing for 20 to 30 days at $25^{\circ}C$ and the LED lights in mixture of white and blue was good for growth period. For effective growth, the temperature, humidity and aeration control in every step was important.

• Journal of Microbiology and Biotechnology
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• v.18 no.1
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• pp.183-187
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• 2008

Vascular endothelial growth factors (VEGFs) are a family of proteins that mediate angiogenesis. $VEGF_{165}$ is a VEGF-A isoform and has been extensively studied owing to its potential use in therapeutic angiogenesis. This study established Chinese hamster ovary (CHO) cells overexpressing recombinant human $VEGF_{165}$ $(rhVEGF_{165})$ protein. The production rate of the established CHO cells was over 80mg/l of $rhVEGF_{165}$ protein from a 7-day batch culture process using a 7.5-l bioreactor with a 5-l working volume and serum-free medium. The $rhVEGF_{165}$ protein was purified to homogeneity from the culture supernatant using a two-step chromatographic procedure that resulted in a 48% recovery rate. The purified $rhVEGF_{165}$ protein was a glycosylated homodimeric protein with a higher molecular weight (MW) than the protein expressed from insect cells, suggesting that the glycosylation of the $rhVEGF_{165}$ protein in CHO cells differed from that in insect cells. The purified $rhVEGF_{165}$ protein in this study was functionally active with a half-maximal effective concentration of 3.8ng/ml and specific activity of $2.5{\times}10^5U/mg$.

• Journal of Plant Biotechnology
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• v.5 no.1
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• pp.1-6
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• 2003

The adventitious root of Panax ginseng C.A. Meyer is regarded as an efficient alternative to cell culture or hairy root culture for biomass production due to its fast growth and stable metabolite production. To determine optimal culture conditions for the bioreactor culture of ginseng roots, experiments have been conducted on physical and chemical factors such as bioreactor type, dissolved oxygen, gas supply, aeration, medium type, macro- and micro-elements, medium supplement during culture period, sucrose concentration, osmotic agents, medium pH and light. Elicitation is a key step to increase ginsenoside accumulation in the adventitious roots but biomass growth is severely inhibited by elicitor treatment. To obtain high ginsenoside content with avoiding biomass decrease, we applied two-stage bioreactor culture system. Ginseng adventitious roots were cultured for 40 days to maximize biomass increase followed by elicitation for 7 days to enhance ginsenoside accumulation. We also experimented on types and concentrations of jasmonate to determine optimal elicitation methods. In this paper, we discussed several factors affecting the root propagation and ginsenoside accumulation. Based on the results obtained from previous experiments we have established large-scale bioreactor system (1 ton-10 ton) for the efficient production of ginseng adventitious roots and bioactive compounds including ginsenoside. Still, experiments are on going in our laboratory to determine other bioactive compounds having effects on diet, high blood pressure, DPPH elimination and increasing memories.