• Journal of Plant Biotechnology
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• 제41권4호
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• pp.236-241
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• 2014

An efficient regeneration protocol for stem disc culture of Populus euramericana, which is important species for bioenergy resource in agroforestry, was established. The number of explants that were obtained and the number of explants that regenerated varied with the genotypes. However, in all the genotypes, stem disc culture produced more regenerated shoots than did in axillary bud culture. A comparison of the effects of cytokinin type and concentration on shoot regeneration in different explants (i.e., petiole, leaf, and root segments of P. euramericana) revealed that a concentration of $0.002mg\;l^{-1}$ thidiazuron (TDZ) used on petiole segments resulted in the greatest shoot regeneration (95.83%). The hormonal requirements for the greatest shoot regeneration in the three explant types varied. Different concentrations of $AgNO_3$ and $CoCl_2$ were added separately to the medium to stop the yellowing and subsequent necrosis of the regenerated shoots. Lower concentrations (3 and $5mg\;l^{-1}$) of these compounds improved shoot regeneration and elongation, compared with the control. The in vitro-regenerated shoots were transferred to rooting medium and subsequently acclimatized. The highly efficient regeneration system of P. euramericana reported here can be used for mass propagation of this recalcitrant for regeneration, economically important tree species.

• 한국가축번식학회지
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• 제20권3호
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• pp.333-341
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• 1996

One of the biggest problems involved in transgenic animal production is lack of appropriate market genes. To overcome this problem, we tested whether the genes of SEAP (secreted alkaline phosphatase) and GFP (green fluorescence protein) on our retrovirus vectors can be applicable to the transgenic animal production. The main advantage of these marker genes over other generally mainpulation can be selected without sacrificing viability. The results obtained in this study are summarized as follows: 1. Removal of zona pellucida from the mouse zygotes did not affect embryo developments to blastocysts. 2. Co-culture of zona-free embryos with virus-producing cells for 6 hours also did not affect embryo developments to blastocysts. 3. Among 58 blastocysts developed from the zona-free zygotes co-cultured with the virus-producing cells, SEAP expression was observed from the 6 blastocysts. 4. Expression of the GFP gene was detected from the virus- producing cells but no embryo expressing the gene was counted among 50 blastocysts developed from the zona-free zygotes co-cultured with the virus-producing cells.

• 한국지하수토양환경학회지:지하수토양환경
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• 제17권5호
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• pp.49-55
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• 2012

A 1.28 L-batch reactor and continuous-flow stirred tank reactor (CFSTR) fed with formate and trichloroethene (TCE) were operated for 120 days and 56 days, respectively, to study the effect of formate as electron donor on anaerobic reductive dechlorination (ARD) of TCE to cis-1,2-dichloroethylene (c-DCE), vinyl chloride (VC), and ethylene (ETH). In batch reactor, injected 60 ${\mu}mol$ TCE was completely degraded in the presence of 20% hydrogen gas ($H_2$) in less than 8 days by anaerobic dechlorination mixed-culture (300 mg-soluble protein), Evanite Culture with ability to completely degrade tetrachloroethene (PCE) and -TCE to ETH under anaerobic conditions. Once the formate was used as electron donor instead of hydrogen gas in batch or chemostat system, the TCE-dechlorination rate decreased and acetate production rate increased. It indicates that the concentration of hydrogen produced in both systems is possibly more close to threshold for homoacetogenesis process. Soluble protein concentration of Evanite culture during the batch test increased from 300 mg to 688 mg for 120 days. Through the protein monitoring, we confirmed an increase of microbial population during the reactor operation. In CFSTR test, TCE was fed continuously at 9.9 ppm (75.38 ${\mu}mol/L$) and the influent formate feed concentration increased stepwise from 1.3 mmol/L to 14.3 mmol/L. Injected TCE was accumulated at 18 days of HRT, but TCE was completely degraded at 36 days of HRT without accumulation of the injected-TCE during the left of experiment period, getting $H_2$ from fermentative hydrogen production of injected formate. Although c-DCE was also accumulated for 23 days after beginning of CFSTR operation, it reached steady-state in the presence of excessive formate. We also evaluated microbial dynamic of the culture at different chemical state in the reactor by DGGE (denaturing gradient gel electrophoresis).

• 한국가축번식학회지
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• 제23권4호
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• pp.337-345
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• 1999

본 연구는 소 체외수정란의 체외배양 시 공동배양배지 및 첨가되는 단백질원에 따른 소 체외수정란의 체외발육능을 검토하였다. 소의 미성숙 난포란을 체외에서 성숙, 수정시킨 후, BSA 또는 FBS를 첨가한 TCM-199 또는 CR1aa 배양액으로 단순배양 또는 난구세포, 소 난관상피세포 (BOEC) 및 Buffalo rat 간세포 (BRLC)와의 공동배양 후, 체외수정란의 분할율 및 발육능을 검사하였다. 소 성숙 난포란을 체외수정 후, 분할율은 배양액의 종류에 관계없이 BSA를 첨가한 경우에 유의적으로 높았다 (P＜0.01). 분할된 수정란을 BSA 또는 FBS가 첨가된 TCM-l99 또는 CRlaa 배양액 내에서 단순배양한 결과, 배반포 발육율은 단백질원에 관계없이 CR1aa 액에서 배양한 경우가 유의적으로 높았다 (P＜0.05). 분할된 수정란을 난구세포 또는 BOEC와 공동배양 시, TCM-l99 배양액에서는 FBS에 비하여 BSA 첨가구가 높은 배반포 형성율을 보였으나 (P＜0.05), CRlaa 배양액에서는 BSA와 FBS 첨가구 모두 높은 발육율이 얻어졌다. 한편, 분할된 수정란을 BRLC의 단층세포와 공동배양 시에는 배양액의 종류와 관계없이 BSA에 비하여 FBS가 수정란의 발육율을 향상시켰다 (P＜0.05). 본 실혐의 결과는 배양액 중에 BSA 첨가가 소 체외수정란의 분할을 촉진할 수 있으며, 체외수정란을 체세포와 공동배양 시, 수정란의 발육율이 배양액 및 첨가 단백질원의 종류에 따라 영향을 받아, TCM-199 액에서 난구세포 또는 BOEC와 공동 배양하는 경우에는 BSA 첨가가 효과적일 수 있음을 수 있음을 보여준다.

• 대한수의학회지
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• 제38권2호
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• pp.359-365
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• 1998

This study was conducted to isolate the protozoan parasite Babesia gibsoni by intraerythrocytic culture method of micoraerophilous stationary phase(MASP) and evaluate the possibility of application for the detection of B gibsoni in canine babesiosis. Also, indirect fluorescent antibody test(IFAT) and thick blood smear(giemsa stain), direct light microscopy (DLM), as control diagnostic tests, were conducted to compare diagnostic effects between MASP, IFAT and DLM. The results obtained from this study were summarized as follows. The protozoan parasite B gibsoni multiplied in 24-well polystyrene plate containing 1.2ml of canine red blood cell suspension in RPMI 1640 medium(pH 7.0) which is contained 20~40% normal canine serum(NCS) under the MASP condition of 5% $CO_2$ and 95% air at $37^{\circ}C$ incubator. Under the above MASP culturing system the percentage of parasitized erythrocytes(PPE) after incubation for 9 days reached the peak. The levels of PPE in MASP culture were shown more higher by exchanging the medium at 24 hour intervals. The parasite were purely isolated from MASP culture of canine red blood cells collected from dogs(pit bullterrier) infected with B gibsoni naturally. Among the total of 83 heads of pit bullterrier blood samples the positive rate was 32 heads(38.5%) in DLM, 45 heads(54.2%) in IFAT and 42 heads(50.6) in MASP culture. In negative cases of IFAT and DLM the isolation rates of B gibsoni by MASP culture were 16 heads(42.1%) of 38 heads and 16 heads(28.6%)% of 56 heads, respectively. From this study it was suggested that MASP culture method by RPMI 1640 medium was a reliable and useful diagnostic test for the diagnosis of B gibsoni infections in canine babesiosis.

• 한국산림과학회지
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• 제97권6호
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• pp.650-655
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• 2008

유칼리나무 펠리타(Eucalyptus pellita) 선발목(5년생)의 기내 대량증식을 위해 1L 규모로 1) 고체배양(대조구), 2) 액체정치배양, 3) 개방형 액체배양 등 3가지 배양법이 시도 되었고, 그 결과를 토대로 배양규모를 10L로 확대하여 대량배양 가능성을 조사하였다. 1L 배양용기에서 4주간 배양한 결과, 공기가 공급 된 개방형 액체배양에서 가장 좋은 생장 결과를 얻었다. 이는 액체배지에 의한 원활한 염류 공급과 배양용기내 공기의 환기에 의해 얻어진 결과로 생각된다. 배양 규모를 10L로 확대하여 액체대량배양을 실시한 결과, 식물체의 생장은 대조구인 고체배양에 비교하여 마디 수에 있어서 370%, 엽 면적 3.6배, 그리고 신초 길이 3.3배 신장하는 결과를 보였다. 이는 개방형 액체대량배양 시스템이 유칼리나무 클론증식시 생산성을 향상시키는데 적합한 시스템임을 보여주는 결과라고 하겠다.

• 한국멀티미디어학회논문지
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• 제18권11호
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• pp.1319-1331
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• 2015

In the Republic of Korea, there's been a culture called 'Hyo' since Koryo Dynasty and this word represents the meaning of paying utmost respects to one's own parents and ancestors whether they are alive or have passed. However, nowadays, most of people live away from their family gravesites so that they do not and cannot take care of them except on the special holidays. For this reason, people could not respond promptly to the incidents occurred at the sites as they receive notifications much later dates most of the time. Thus, in this paper, we propose a low-cost gravesite monitoring system which the users can immediately respond to the disastrous events after being informed of current situations through PLC without delay. For the performance evaluation, the lab and test bed experiments were performed on an actual ship using 200Mbps and 500Mbps products instead of performing an on-site experiment after the system has actually been constructed. The Mountain Region PLC was installed on the power lines and the result showed successful 36.14Mbps communication. Therefore, we expect that this study will contribute in time and cost reduction while constructing the internet infrastructures in mountain regions or building the Smart-graves, tumulus, and charnel houses.

• 미생물학회지
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• 제44권2호
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• pp.130-134
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• 2008

미생물 자동배양기를 이용한 감염성 물질의 검출은 시험적 오차의 경감은 물론 분리율 향상과 시간 단축을 가능하게 하였다. BacT/ALERT 3D 자동배양기는 미생물성장 시 발생되는 이산화탄소를 비색법으로 검출하는 장비로서 임상시료를 이용한 미생물 배양과 검출에 이용되어왔다. 자동배양기의 효율성을 검증하고 의약품의 무균 시험에 적용가능한지를 평가하기 위하여 6종의 세균을 이용하여 중식 및 검출특성을 분석하였다. 3종의 호기성세균 Pseudomonas aeruginosa, Micrococcus Iuteus, Bacillus subtilis와 Staphylococcus aureus는 1 CFU가 31.44 시간 내에 검출되었고, 혐기성균인 Clostridium sporogenes는 15.96시간에 균의 중식이 감지되었다. 저성장 혐기성세균인 Propionibacterium acnes는 $10^4$ CFU의 균수에서 검출에 129.36시간이 소요되었다. 이 같은 결과는전통적 직접 배양법보다 검출감도에 있어서 $10\sim10^5$배 높고 검출시간을 $2\sim10$10시간 단축한 것으로서 자동배양법이 미생물 증식과 검출에 효율적임을 말해준다. 따라서 본 시험에서 이용한 자동배양법은 임상시료에서의 감염원 진단뿐 아니라 생물의약품의 무균시험에 이용 가능하다고 판단된다.

• 대한화장품학회지
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• 제47권4호
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• pp.297-304
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• 2021

본 연구에서는 미백 기능성 화장품 및 원료의 효능을 평가하는 동물대체시험법을 개발하기 위하여 세포수준과 색소화피부모델(KeraSkin-M^TM)에서 기존에 잘 알려진 4종의 미백기능성원료(arbutin, ascorbic acid, kojic acid, niacinamide)의 효능을 평가하였다. 특히 기존 시험법의 보완을 위해 인체피부유래 케라틴세포와 멜라닌세포를 혼합하고 공배양하여 색소화피부모델을 제작하였다. 그 결과 색소화피부모델을 이용하여 미백효능을 평가함으로써 세포수준에서는 확인이 어려웠던 각 피부세포층에 따른 멜라닌 과립과 멜라닌캡(melanin cap)의 분포 등의 지표들을 추가로 확인할 수 있었으며 이미지분석을 통한 정량화로 음성대조군 대비 통계적 유의성을 확인할 수 있었다. 이러한 결과는 KeraSkin-M^TM을 이용한 미백효능평가법이 기존에 사용하던 총멜라닌함량와 타이로시나아제 저해 평가를 보완할 수 있는 새로운 평가법으로 사용할 수 있음을 시사한다.

• Journal of Microbiology and Biotechnology
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• 제30권5호
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• pp.770-776
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• 2020

Genistein is a type of isoflavonoid found predominantly in leguminous plants. Genistein has diverse biological activities, such as anthelmintic and antioxidant effects, as well as inhibitory effects on the growth of several cancers. In addition, genistein is well known as a phytoestrogen. In this study, we attempted to biologically synthesize genistein from either p-coumaric acid or naringenin using Escherichia coli as a biotransformation host. Four genes, Os4CL, PeCHS, RcIFS, and OsCPR, were used for genistein production. To functionally express RcIFS and OsCPR, two members of the cytochrome P450 family, in E. coli, the membrane-binding anchor domain of each gene was removed, and RcIFS and OsCPR were translationally fused to generate an RcIFS-OsCPR hybrid. Os4CL and PeCHS, or the RcIFS-OsCPR hybrid, were then transformed into E. coli BL21(DE3). Using these strains, we optimized our culture system at a laboratory scale in terms of the cell density, concentrations of substrate and isopropyl-β-D-thiogalactoside, temperature, and culture medium. Under the optimized culture conditions, genistein was produced at up to 35 mg/l and 18.6 mg/l using naringenin and p-coumaric acid, respectively.