• 제목/요약/키워드: cell yield

검색결과 1,255건 처리시간 0.031초

Optimal Conditions for the Expression of Glycoprotein E2 of Classical Swine Fever Virus using Baculovirus in Insect Cells

  • Bae, Sung Min;Lee, Seung Hee;Kwak, Won Suk;Ahn, Yong Oh;Shin, Tae Young;Woo, Soo Dong
    • International Journal of Industrial Entomology and Biomaterials
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    • 제29권2호
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    • pp.207-213
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    • 2014
  • The structural proteins of classical swine fever virus (CSFV) consist of nucleocapsid protein C and envelope glycoprotein $E^{rns}$ (E0), E1 and E2. Among them, E2, the most immunogenic of the CSFV glycoproteins, induces a protective immune response in swine. In this study, to determine the optimal expression conditions of glycoprotein E2 using baculovirus system, we investigated the influence of insect cells and media to the expression of recombinant E2. Recombinant virus containing glycoprotein E2 coding gene was constructed with bApGOZA DNA. Expression of the glycoprotein E2 was analyzed by SDS-PAGE and Western blot analysis using anti-CSFV E2 monoclonal antibodies. Expression of glycoprotein E2 in Sf21 cells was first observed after 3 days and reached a maximum on the 5th day after infection. Furthermore, the highest levels of glycoprotein E2 expression were observed at multiplicity of infection (MOI) of 5. When three different insect cell lines (Sf21, High-Five and Se301) were tested, High-Five cells showed the highest production. In addition, four different serum-free and serum-supplemented media, respectively, were tested for the expression of glycoprotein E2 and the budded virus (BV) titers. As a result, serum-supplemented medium provided the best conditions for protein production and the BV yield.

뽕나무버섯(Armillaria mellea) 균사체 생산의 최적화 (Optimal Condition for Mycelial Production of Armillaria mellea)

  • 김명곤;최한석;박효숙;김성준
    • 한국균학회지
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    • 제31권3호
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    • pp.187-191
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    • 2003
  • 뽕나무 버섯균은 식용버섯으로써 노인병, 중풍, 현기증, 두통, 신경쇠약, 불면증, 사지마비, 소아마비, 간질 등에 효과가 있는 것으로 알려져 있으므로, 뽕나무 버섯 균사체의 고농도 배양을 위한 최적 조건확립 하고자 하였다. 뽕나무 버섯균사체 생산의 최적배지 선정을 위하여 7종의 배지를 대상으로 수행한 결과 맥아즙배지가 뽕나무 버섯 균사체 생산에 가장 우수하였다. 맥아즙 배지에서 뽕나무 버섯균 균사체 생산을 위한 적정조건은 맥아즙 농도 $15\;Brix^{\circ}$이었으며 온도, pH, 배양기간의 조건을 반응표면 분석법에 의해서 실행한 결과, $25.9^{\circ}C$, pH 5.72, 15.22일의 배양에서 3,40 g/50 ml의 최대 예측치를 나타내었다. 이는 실제 실험결과치와도 대체로 일치하는 경향이었다.

Transcriptome analyses of the ginseng root rot pathogens Cylindrocarpon destructans and Fusarium solani to identify radicicol resistance mechanisms

  • Li, Taiying;Kim, Jin-Hyun;Jung, Boknam;Ji, Sungyeon;Seo, Mun Won;Han, You Kyoung;Lee, Sung Woo;Bae, Yeoung Seuk;Choi, Hong-Gyu;Lee, Seung-Ho;Lee, Jungkwan
    • Journal of Ginseng Research
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    • 제44권1호
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    • pp.161-167
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    • 2020
  • Background: The ascomycete fungi Cylindrocarpon destructans (Cd) and Fusarium solani (Fs) cause ginseng root rot and significantly reduce the quality and yield of ginseng. Cd produces the secondary metabolite radicicol, which targets the molecular chaperone Hsp90. Fs is resistant to radicicol, whereas other fungal genera associated with ginseng disease are sensitive to it. Radicicol resistance mechanisms have not yet been elucidated. Methods: Transcriptome analyses of Fs and Cd mycelia treated with or without radicicol were conducted using RNA-seq. All of the differentially expressed genes (DEGs) were functionally annotated using the Fusarium graminearum transcript database. In addition, deletions of two transporter genes identified by RNA-seq were created to confirm their contributions to radicicol resistance. Results: Treatment with radicicol resulted in upregulation of chitin synthase and cell wall integrity genes in Fs and upregulation of nicotinamide adenine dinucleotide dehydrogenase and sugar transporter genes in Cd. Genes encoding an ATP-binding cassette transporter, an aflatoxin efflux pump, ammonium permease 1 (mep1), and nitrilase were differentially expressed in both Fs and Cd. Among these four genes, only the ABC transporter was upregulated in both Fs and Cd. The aflatoxin efflux pump and mep1 were upregulated in Cd, but downregulated in Fs, whereas nitrilase was downregulated in both Fs and Cd. Conclusion: The transcriptome analyses suggested radicicol resistance pathways, and deletions of the transporter genes indicated that they contribute to radicicol resistance.

맥주 공장 부산물 효모의 최적 자가 소화 조건 결정 (Optimization for Autolysis of Brewers Yeast Slurry)

  • 손상목;김재식
    • 한국식품과학회지
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    • 제35권2호
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    • pp.201-205
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    • 2003
  • 맥주 공장 부산물 효모의 용도를 다양화하기 위하여 효모 슬러리를 자가 소화시키고 상등액을 농축하여 조미료로 이용할 수 있는 효모추출물을 제조하였다. 맥주 효모의 세포벽에 부착되어 있는 호프 쓴맛 성분을 제거하기 위하여 효모 슬러리에 20%(w/v) NaOH 용액을 가하여 pH 9.8로 맞추고, 약 5분간 방치한 후 즉시 원심분리하면서 물로 세척하였다. 세척 후 효모 쓴맛 정도는 24.1 bitterness unit(BU)로서 threshold value 25.0 BU 미만이었다. 세척한 효모를 자가 소화시킨 후 수율[(수득한 효모추출물의 고형분 무게)/(투입 효모슬러리의 고형분 무게)${\times}100$]을 조사한 결과, 효모슬러리 농도를 $9{\sim}10%(w/w)$로 하고, 온도 $53^{\circ}C$, pH 6.8에서 20시간 이상 자가 소화시키면 34%(w/w)이상, 최대 38%(w/w)까지의 수율로 효모 추출물을 얻을 수 있었다.

A Modified Process for Producing High Quantities of Bio-Germanium in Yeast and a Study of Its Oral Toxicity

  • Park, So-Young;Joo, Seong-Soo;Won, Tae-Joon;Chung, Jin-Woong;Lee, Sung-Hee;Oh, Sun-Woo;Lee, Do-Ik;Hwang, Kwang-Woo
    • Food Science and Biotechnology
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    • 제16권1호
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    • pp.78-82
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    • 2007
  • Since germanium has been shown to be beneficial for the treatment of diseases such as cancer and rheumatic arthritis, we developed an adapted process of bio-germanium preparation using inorganic germanium. In the present study we determined the optimal conditions for culturing yeast Saccharomyces cerevisiae (KCTC-1199), and the best concentrations of inorganic germanium for the adaptation process. The resulting method was successful at producing high quantities of germanium yeasts. The following are the culture conditions that obtained the highest level of productivity: an inorganic germanium concentration of 3,000-5,000 ppm, a pH of 6.5, a temperature of $35^{\circ}C$, and 20 hr of incubation time. In addition to this high-yield quantity study, we observed the acute oral toxicity of mice treated with Geranti Bio-Ge $Yeast^{(R)}$. We found no changes in body weight, or in the mortality between the control groups and the bio-germanium yeast group. There were also no digestive problems such as diarrhea that occurred in either group.

배지조성이 Gluconacetobacter hansenii PJK의 Bacterial Cellulose의 교반 생산에 미치는 영향 (Medium Composition Affecting Production of Bacterial Cellulose by Gluconacetobacter hansenii PJK in an Agitated Culture)

  • 정재용;장호남;박중곤
    • KSBB Journal
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    • 제19권6호
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    • pp.451-456
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    • 2004
  • 교반배양시 배지에 첨가되는 에탄올의 농도가 증가할수록 $Cel^-$ mutant의 발생율이 증가하므로 $1.5\%$ 에탄올이 첨가된 배지의 경우 500 rpm보다 높은 교반속도를 유지해야 효과적으로 $Cel^-$ mutant의 발생이 억제되고 BC 생산량을 증가시킬 수 있다. 교반 배양과 같은 강한 shear stress의 배양환경 하에서는 진탕배양의 경우와 달리 $1.0\%$ 에탄올을 분할공급하더라도 BC 생산의 효율성을 높일 수 없었다. Phosphate ion이 첨가된 배지보다 첨가되지 않은 배지에서 BC 생산량이 더 높았다. 배지조성 중 아세트산의 농도를 $1.0\%$로 증가시키면 균체의 건조중량은 변화가 없었으나 BC 생산량이 감소되었다. 배양액의 pH 제어는 BC의 생산에는 영향을 미치지 않았으나 균체의 성장과 WSPS의 생산을 향상시켰다.

Purification of Capsular Polysaccharide Produced by Streptococcus pneumoniae Serotype 19A

  • Jung, Seung-Jin;Seo, Eun-Seong;Yun, Sang-Il;Minh, Bui Nguyet;Jin, Sheng-De;Ryu, Hwa-Ja;Kim, Do-Man
    • Journal of Microbiology and Biotechnology
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    • 제21권7호
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    • pp.734-738
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    • 2011
  • Streptococcus pneumoniae is a major cause of invasive infection in young infants and older adults. There are currently 90 capsular serotypes identified and 23 serotypes (1, 2, 3, 4, 5, 6B, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19F, 19A, 20, 22F, 23F, and 33F) are responsible for about 90% of invasive disease. Among the more than 90 different S. pneumoniae serotypes, serotype 19A is globally very prevalent. A simplified purification procedure including adjustment of cell lysate pH to 4.5, fractionation with 50. 80% ethanol, and dialysis rendered capsular polysaccharide (CPS) in a yield of $31.32{\pm}3.11$ mg from 1 l culture (75% recovery after lyses). The product contained only 69.6 ${\mu}g$ of protein (99.78% purity) and 0.8mg (sum of the precipitants from 50~60%, 60~70%, and 70~80%) of nucleic acid (97.45% purity). The purified CPS was conjugated with bovine serum albumin; the product size ranged from 100 to 180 kDa.

Dynamics of Microalgae Along the Coastal Areas of Sooyoung Bay, Busan, South Korea

  • Prasad, Binod;Thiyam, General;Lee, Dong-Gyu;Kim, Moo-Sang;Cho, Man-Gi
    • 한국해양바이오학회지
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    • 제5권4호
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    • pp.40-45
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    • 2011
  • Microalgae are one of the major, sustaining components of ecosystem processes and are responsible for biogeochemical reactions that drive our climate changes. Despite this, many marine microalgae are poorly described and little is known of their abundance and distribution along the coastal areas of Sooyoung Bay, Busan, South Korea. The present study has been conducted from November, 2011 to August, 2009 with the objective to provide an overview of the taxonomy diversity and abundance of microalgae along the coastal areas of the Sooyoung Bay. Water samples were collected from different sites, which were located by using a GPS tracker. Chlorophyll fluorescence of the water samples were measured by using ToxY-PAM dual-channel yield analyzer. The chlorophyll fluorescence values were relatively higher during the spring and summer and even in the region near to the sea port. Similarly the abundance of microalgae was higher near the port but diversity index had lower values. The temperature and pH values were same at all the sites. However, only the temperature varied during the sampling period, with higher values during summer and lower in winter. From the preliminary results, the following class of microalgae were found; Bacillariophyceae, Dinophyceae, Silicoflagellate and Cryptophyceae. With a future ongoing work, microalgae are being isolated to establish single cell culture and for identification using light microscopic observations, photography and molecular approaches.

Characterization of Homologous Defective Interfering RNA during Persistent Infection of Vero Cells with Japanese Encephalitis Virus

  • Yoon, Sung Wook;Lee, Sang-Yong;Won, Sung-Yong;Park, Sun-Hee;Park, Soo-Young;Jeong, Yong Seok
    • Molecules and Cells
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    • 제21권1호
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    • pp.112-120
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    • 2006
  • It has been suggested that defective interfering (DI) RNA contributes to the persistence of Japanese encephalitis virus (JEV). In this study, we characterized molecular and biological aspects of the DI RNA and its relation to viral persistence. We identified a homologous DI virus intimately associated with JEV persistence in Vero cells. The production of DI RNA during undiluted serial passages of JEV coincided with the appearance of cells refractory to acute infection with JEV. We also established a Vero cell clone with a persistent JEV infection in which the DI RNA coreplicated efficiently at the expense of helper virus. The infectious virus yield of the clone fluctuated during its growth depending upon the amount of DI RNA accumulated in the previous replication cycle. Identification of the corresponding negative-sense RNA of the DI RNA indicated that the DI RNA functioned as a replication unit. Most of the DI RNA molecules retained their open reading frames despite a large deletion, encompassing most of the prM, the entire E, and the 5' half of the NS1 gene. Taken together, these observations suggest that the generation of homologous DI RNA during successive JEV acute infections in Vero cells probably participates actively in persistent JEV infection.

병원성사상균에 의한 Tall Fescue ( Festuca arundinacea Schreb. ) 의 수량 손실과 길항 미생물에 의한 그 방제 (Yield Decrease of Tall Fescue ( Festuca arundinacea Schreb. ) by Pathogenic Fungi and its Control by Antagonistic Bacteria)

  • 최기춘;송채은;이종경;김종현;이영환;윤창
    • 한국초지조사료학회지
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    • 제18권2호
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    • pp.133-142
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    • 1998
  • This study was conducted to investigate the effects of antagonistic bacteria and pathogenic fungi on the growth of tall fescue(Festuca arundinacea Schreb.) in continuous cropping soil(CCS) and non-continuous cropping soil(NCCS). Tall fescue was established by seeding into pots of 11 cm in diameter and 9 cm in depth containing 1 : 1 mixture of soil and vermiculite, and cultivated at pots with antagonistic bacteria and pathogenic fungi in a vinyl house. The bacteria used in this study were Bacillus subtilis and hsants. B. subtilis was isolated and identified kern forage rhizosphere soil and fusants were isolated through cell hsion from B. subtilis and B. thwingiensis. B. subtilis was named as B. subtilis 101 and hsants were named as F-3, F-7 and F-8. In dark culture experiment, tall fescue inoculated with the antagonistic bacteria lived longer than that of control in both CCS and NCCS. However, tall fescue of CCS lived shorter than that of NCCS. Dry weight of tall fescue inoculated with the antagonistic bacteria was higher than that of tall fescue inoculated with pathogenic hngi in both CCS and NCCS(P< 0.05), and the antagonistic bacteria showed positive effects on the growth of tall fescue. However, Dry weight of tall fescue was decreased by the inoculation of the pathogenic b g i in both CCS and NCCS(P< 0.05).

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