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Functional Expression of an Anti-GFP Camel Heavy Chain Antibody Fused to Streptavidin (Streptavidin이 융합된 GFP항원 특이적인 VHH 항체의 기능적 발현)

  • Han, Seung Hee;Kim, Jin-Kyoo
    • Journal of Life Science
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    • v.28 no.12
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    • pp.1416-1423
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    • 2018
  • With strong biotin binding affinity ($K_D=10^{-14}M$), the tetrameric feature of streptavidin could be used to increase the antigen binding activity of a camel heavy chain (VHH) antibody through their fusion, here stained with biotinylated horseradish peroxidase and subsequent immunoassays ELISA and Western blot analysis. For this application, we cloned the streptavidin gene amplified from the Streptomyces avidinii chromosome by PCR, and this was fused to the gene of the 8B9 VHH antibody which is specific to green fluorescent protein (GFP) antigens. To express a soluble fusion protein in Escherichia coli, we used the pUC119 plasmid-based expression system which uses the lacZ promoter for induction by IPTG, the pelB leader sequence at the N-terminus for secretion into the periplasmic space, and six polyhistidine tags at the C-terminus for purification of the expressed proteins using an $Ni^+$-NTA-agarose column. Although streptavidin is toxic to E. coli because of its strong biotin binding property, this soluble fusion protein was expressed successfully. In SDS-PAGE, the size of the purified fusion protein was 122.4 kDa in its native condition and 30.6 kDa once denatured by boiling, suggesting the tetramerization of the monomeric subunit by non-covalent association through the streptavidin moiety fusing to the 8B9 VHH antibody. In addition, this fusion protein showed biotin binding activity similar to streptavidin as well as GFP antigen binding activity through both ELISA and Western blot analysis. In conclusion, the protein resulting from the fusion of an 8B9 VHH antibody with streptavidin was successfully expressed and purified as a soluble tetramer in E. coli; it showed both biotin and GFP antigen binding activity suggesting the possible production of a tetrameric and bifunctional VHH antibody.

Vegetation Structure and Growth Environmet of Diabelia spathulata (Siebold & Zucc.) Landrein Population in Mt. Cheonseong, Korea (천성산 주걱댕강나무 개체군의 식생구조와 생육환경)

  • Yi, Myung Hoon;Yoo, Sung Tae;Jang, Jeong Gul
    • Korean Journal of Plant Resources
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    • v.34 no.4
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    • pp.346-361
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    • 2021
  • The range of D. spathulata identified in this survey was between N 35° 24' 58" ~ N 35° 26' 35", E 129° 05' 43" ~ E 129° 07' 04". It is located at an altitude of 98~592 m. The soil pH was strongly acidic in the range of 4.2~4.9, with a canopy openness of 18.56% and a chlorophyll index of 36.74 ± 2.80. As a result of the TWINSPAN analysis, 20 plots of 100 m2 each were divided in 4 communities: Pinus densiflora community, Quercus monglica-Diabelia spathulata community, Quercus serrata-Diabelia spathulata community and Carpinus tschonoskii subassociation. The result of species diversity was 0.7615, and evenness and dominance were found to be 0.6077 and 0.3923, respectively. The height of D. spathulata is up to 3.4 m, and the average height is 1.1 m, with most of the species distributed as shrubbery and herbaceous. The average population density of the 20 plots was 1.635 individuals/m2, the height range of flowering was 1.0 ~ 1.8 (aver. 1.39 m) and the rate of flowering was 27.37%. It's propagation pattern was mainly formed by extending the rhizome to the side, creating a colony of ground stems.

Effects of Dry Heat Treatment on the Reduction of Main Food-Borne Bacteria on Alfalfa Seeds (건열처리를 이용한 알팔파의 주요 식중독균 저감화)

  • Hong, Soon-Young;Kim, Su-jin;Bang, Woo-Suk
    • Journal of Food Hygiene and Safety
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    • v.37 no.4
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    • pp.225-231
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    • 2022
  • In this study, the conditions of dry heat treatment (21 days at 65℃, 16 days at 70℃, 10 days at 75℃, and 7 days at 80℃) were investigated to inactivate Bacillus cereus ATCC 12480, Listeria monocytogenes ATCC SSA81, Staphylococcus aureus ATCC 6538, Escherichia coli O157:H7 ATCC 43894, and Salmonella Typhimurium ATCC 14028 on alfalfa seeds, without affecting the rate of germination of seeds. Alfalfa seeds were inoculated at levels of 6-7 log CFU/g and treated with dry heat at 65℃, 70℃, 75℃, and 80℃; thereafter, the rate of seed germination was determined. The rate of germination was set at 70%, according to the market standards. The bacteria were inactivated when B. cereus was treated with dry heat for 21 days at 65℃, 18 days at 70℃, 14 days at 75℃, and 4 days at 80℃; L. monocytogenes was treated for 21 days at 65℃, 18 days at 70℃, 12 days at 75℃, and 7 days at 80℃; S. aureus was treated for 18 days at 65℃, 18 days at 70℃, 11 days at 75℃, and 4 days at 80℃; E. coli O157:H7 was treated for 21 days at 65℃, 18 days at 70℃, 12 days at 75℃, and 6 days at 80℃; and Sal. Typhimurium was treated for 24 days at 65℃, 22 days at 70℃, 14 days at 75℃, and 7 days at 80℃. For all bacteria, the D-value (R2 = 0.5656-0.7957) significantly decreased when the temperature increased from 65℃ to 80℃ (P<0.05). Since dry heat treatment of alfalfa seeds at 80℃ for 7 days affects their germination rate, dry heat treatment at 75℃ for 14 days is the most effective way to ensure their safety. This study suggests a potential method of bacterial inactivation using dry heat treatment to increase the microbiological safety of sprouts.

Optimization of Ingredients for the Preparation of Chinese Quince (Chaenomelis sinensis) Jam by Mixture Design (모과잼 제조시 혼합물 실험계획법에 의한 재료 혼합비율의 최적화)

  • Lee, Eun-Young;Jang, Myung-Sook
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.7
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    • pp.935-945
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    • 2009
  • This study was performed to find the optimum ratio of ingredients in the Chinese quince jam. The experiment was designed according to the D-optimal design of mixture design, which included 14 experimental points with 4 replicates for three independent variables (Chinese quince paste $45{\sim}60%$, pectin $1.5{\sim}4.5%$, sugar $45.5{\sim}63.5%$). A mathematical analytical tool was employed for the optimization of typical ingredients. The canonical form and trace plot showed the influence of each ingredient in the mixture against final product. By use of F-test, sweetness, pH, L, b, ${\Delta}E$, and firmness were expressed by a linear model, while the spreadmeter value, a, and sensory characteristics (appearance, color, smell, taste, and overall acceptability) were by a quadratic model. The optimum formulations by numerical and graphical method were similar: Chinese quince paste 54.48%, pectin 2.45%, and sugar 53.07%. Optimum ingredient formulation is expected to improve use of Chinese quince and contribute to commercialization of high quality Chinese quince jam.

Significance of Random Urine ${\beta}_2$-Microglobulin as a Marker for Vesico-ureteral Reflux in Children with Urinary Tract Infection (요로감염 환자에서 방광요관역류의 지표로서 무작위 추출뇨중 ${\beta}_2$-microglobulin의 의의)

  • Kim Ji-Hong;Kim Pyung-Kil
    • Childhood Kidney Diseases
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    • v.3 no.1
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    • pp.72-79
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    • 1999
  • Renal scarring associated with vesico-ureteral reflux(VUR) is one of the major causes of end stage renal failure and renal hypertension in children. Urinary ${\beta}_2$-microglobulin(MG) has been suggested as a potential marker for presence of renal tubular damage. This study was designed to evaluate the significance of random urine ${\beta}_2$-MG as a predictor of presence of vesico-ureteral reflux in children with urinary tract infection(UTI). 57 children with urinary tract infection were studied. Patients were devided into two groups; 35($78.9\%$) children have UTI without VUR and 12($21.1\%$) children have UTI and VUR. Beta2-MG and creatinine in random urine sample was measured to decide the excretion ratio(${\beta}_2$-MG/creatinine). Among the 57 children with UTI, 44 children were confirmed by urine culture study and 13 children suspected by compatible clinical feature. Random urine ${\beta}_2$-MG of VUR group ($2.2{\pm}5.91$ mg/L) were significantly higher than that of simple UTI group($0.19{\pm}0.16mg/L$)(P=0.03). The ${\beta}_2$-MG/creatinine ratio of VUR group($32.41{\pm}25.7$) were significantly higher than that of simple UTI group($3.93{\pm}3.44$)(P=0.007). In conclusion, random urine ${\beta}_2$-MG and excretion ratio deserved early predictor of presence of VUR in children with UTI. And this method was more simple and inexpensive than the method of measuring ${\beta}_2$-MG with 24 hour urine collection, so might be a useful screening test for VUR in children with UTI.

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Changes in Moisture Contents of Rice-hull Based Root Media and Growth Responses of 'Seolhyang' Strawberry during Vegetative Propagation (육묘 과정 중 포트에 충진된 팽연왕겨 혼합상토의 함수량 변화와 '설향' 딸기의 생장 반응)

  • Park, Gab Soon;Kim, Yeoung Chil;Ann, Seoung Won;Kang, Hee Kyoung;Choi, Jong Myung
    • Horticultural Science & Technology
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    • v.33 no.1
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    • pp.47-54
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    • 2015
  • This research was conducted to investigate the changes in moisture retention capacities of expanded rice-hull (ERH)-based root media and their influences on the growth of mother and daughter plants in vegetative propagation of 'Seolhyang' strawberry. The proportion of water at the container capacity of ERH medium was in the range of 20 to 23%. This range was lower than the 60 to 66% of strawberry-specialized medium, the 30 to 34% of soil mother material (SMM) and the 30 to 35% of loamy sand. The moisture content of ERH was reduced to 10 to 12% at 8 hours after irrigation, and there were large variations among replications of ERH medium. Among four kinds of root media formulated to contain ERH, the medium of ERH + coir dust (CD) (55 + 45%, v/v) had 26.5 and 32.5% water contents at 20 and 40 days after irrigation to daughter plants, respectively. The m edia o f ERH + sandy loam (S L) and E RH + S MM showed similar trends i n moisture r etention. The pH and EC i n the all root media tested were in the range of 6.7 to 7.1 and 0.03 to $0.08dS{\cdot}m^{-1}$, respectively. The pHs and ECs measured at 20 and 40 days after irrigation were not significantly different in each root medium. Among the root media formulated to contain ERH, the growth of daughter plants was the highest in the treatment of ERH + SL (55 + 45%, v/v). As the blending rate of coir dust was elevated in the ERH + CD media, moisture retention capacity increased gradually, but the growth of daughter plants became worse even though the medium showed higher moisture retention capacity than other root media tested. The growth of roots and aboveground tissues of daughter plants deteriorated in the root media formulated by blending ERH + perlite (PE) at various ratios. The results of this research suggest the optimum formulations of root media and management of moisture content in raising of strawberry daughter plants when ERH is a component of root media.

Magnetic Tunnel Junctions with AlN and AlO Barriers

  • Yoon, Tae-Sick;Yoshimura, Satoru;Tsunoda, Masakiyo;Takahashi, Migaku;Park, Bum-Chan;Lee, Young-Woo;Li, Ying;Kim, Chong-Oh
    • Journal of Magnetics
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    • v.9 no.1
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    • pp.17-22
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    • 2004
  • We studied the magnetotransport properties of tunnel junctions with AlO and AlN barriers fabricated using microwave-excited plasma. The plasma nitridation process provided wider controllability than the plasma oxidization for the formation of MTJs with ultra-thin insulating layer, because of the slow nitriding rate of metal Al layers, comparing with the oxidizing rate of them. High tunnel magnetoresistance (TMR) ratios of 49 and 44% with respective resistance-area product $(R{\times}A) of 3 {\times} 10^4 and 6 {\times} 10^3 {\Omega}{\mu}m^2$ were obtained in the Co-Fe/Al-N/Co-Fe MTJs. We conclude that AlN is a hopeful barrier material to realize MTJs with high TMR ratio and low $R{\times}A$ for high performance MRAM cells. In addition, in order to clarify the annealing temperature dependence of TMR, the local transport properties were measured for Ta $50{\AA} /Cu 200 {\AA}/Ta 50 {\AA}/Ni_{76}Fe_{24} 20 {\AA}/Cu 50 {\AA}/Mn_{75}Ir_{25} 100 {\AA}/Co_{71}Fe_{29} 40 {\AA}/Al-O$ junction with $d_{Al}= 8 {\AA} and P_{O2}{\times}t_{0X}/ = 8.4 {\times} 10^4$ at various temperatures. The current histogram statistically calculated from the electrical current image was well in accord with the fitting result considering the Gaussian distribution and Fowler-Nordheim equation. After annealing at $340^{\circ}C$, where the TMR ratio of the corresponding MTJ had the maximum value of 44%, the average barrier height increased to 1.12 eV and its standard deviation decreased to 0.1 eV. The increase of TMR ratio after annealing could be well explained by the enhancement of the average barrier height and the reduction of its fluctuation.

THE LORENTZ FORCE IN ATMOSPHERES OF CP STARS: θ AUR

  • VALYAVIN G.;KOCHUKHOV O.;SHULYAK D.;LEE B.-C.;GALAZUTDINOV G.;KIM K.-M.;HAN I.
    • Journal of The Korean Astronomical Society
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    • v.38 no.2
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    • pp.283-287
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    • 2005
  • The slow evolution of global magnetic fields and other dynamical processes in atmospheres of CP magnetic stars lead to the development of induced electric currents in all conductive atmospheric layers. The Lorentz force, which results from the interaction between a magnetic field and the induced currents, may modify the atmospheric structure and provide insight into the formation and evolution of stellar magnetic fields. This modification of the pressure-temperature structure influences the formation of absorption spectral features producing characteristic rotational variability of some spectral lines, especially the Balmer lines (Valyavin et al., 2004 and references therein). In order to study these theoretical predictions we began systematic spectroscopic survey of Balmer line variability in spectra of brightest CP magnetic stars. Here we present the first results of the program. A0p star $\Theta$ Aur revealed significant variability of the Balmer profiles during the star's rotation. Character of this variablity corresponds to that classified by Kroll (1989) as a result of an impact of significant Lorentz force. From the obtained data we estimate that amplitudes of the variation at H$\alpha$, H$\beta$, H$\gamma$ and H$\delta$ profiles reach up to $2.4\%$during full rotation cycle of the star. Using computation of our model atmospheres (Valyavin et al., 2004) we interpret these data within the framework of the simplest model of the evolution of global magnetic fields in chemically peculiar stars. Assuming that the field is represented by a dipole, we estimate the characteristic e.m.f. induced by the field decay electric current (and the Lorentz force as the result) on the order of $E {\~} 10^{-11}$ cgs units, which may indicate very fast (< < $10^{10}$ years) evolution rate of the field. This result strongly contradicts the theoretical point of view that global stellar magnetic fields of CP stars are fossil and their the characteristic decay time of about $10^{10}$ yr. Alternatively, we briefly discuss concurring effects (like the ambipolar diffusion) which may also lead to significant atmospheric currents producing the observable Lorentz force.

Fine-scale Spatial Genetic Structure of a Small Natural Stand of Populus davidiana in South Korea using AFLP markers (AFLP 마커를 이용한 소규모 사시나무림의 공간적 유전구조 구명)

  • Lee, Min Woo;Hong, Kyung Nak;Park, Yu Jin;Lee, Jei Wan;Lim, Hyo In
    • Journal of Korean Society of Forest Science
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    • v.105 no.3
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    • pp.309-314
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    • 2016
  • A locally adapted plant population under harsh environmental changes might survive for a long generation through maintaining proper level of genetic diversity. When it happens losing the genetic diversity too much fast, the population could be declining and probably become extinct. An isolated small population of Populus davidiana was investigated to study out the genetic diversity and the fine-scale spatial genetic structure. The estimated number of adult trees in the population of Mt. Worak, South Korea, was 350 in the total area of $14,000m^2$. The number of adults in a study plot ($70m{\times}70m$) was 123. The average age was 16-year-old and a 32-year-old tree was the oldest. The distribution of individuals was slightly aggregated in the plot. Sixty-one among the 123 individuals were randomly sampled to estimate genetic variation using AFLP markers. One hundred fifty-one (77%) of total 196 amplicons were polymorphic from six AFLP primer combinations. The average number of loci per primer combination was 32.7 (S.D.=7.2). Expected heterozygosity ($H_e$) and Shannon's diversity index (S.I.) were 0.154 and 0.254, respectively. These values were extremely lower than those of other P. davidiana populations in South Korea. Genetic patchiness was showed within 21 meters by spatial autocorrelation analysis and the isolated small size of population might be mainly attributed to the formation of such small patch size.

Analysis of Mineral, Amino Acid and Vitamin Contents of Fruiting Body of Sparassis crispa. (꽃송이버섯의 미네랄, 아미노산, 비타민 함량분석)

  • Shin, Hyun-Jae;Oh, Deuk-Sil;Lee, Hee-Duck;Kang, Hyeong-Bong;Lee, Chul-Won;Cha, Wol-Suk
    • Journal of Life Science
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    • v.17 no.9 s.89
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    • pp.1290-1293
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    • 2007
  • The nutritional composition of fruiting body of Sparassis crispa has been analyzed for medicinal and edible uses. Minerals in S. crispa were found to be as follows; potassium (1,299.44 mg), phosphorus (104.73 mg), sodium (98.21 mg), magnesium (54.86 mg), calcium (8.39 mg), iron (7.61 mg), zinc (6.37 mg), copper (1.31 mg) and manganese (0.63 mg) based on 100 g of mushroom dry weight. In 20 kinds of total amino acids found in S. crispa, sum of glutamic acid and glutamine content was the highest (1,960 mg/l00 g) and sum of aspartic acid and asparagine, tryptophan, leucine and alanine were followed. Concerning free amino acids, glutamic acid, tryptophan, glutamine and aspartic acid were dominant. Among 8 vitamins detected, the vitamin E content was the highest (408.5 mg) based on 100 g of mushroom dry weight, then vitamin C, niacin and pantothenic acid were followed.