• 제목/요약/키워드: PCR Device

검색결과 29건 처리시간 0.034초

Expression and Purification of GFPuv/Cytochrome c-552 Fusion Protein in E. coli

  • 홍을재;이상온;최정우;홍억기
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2003년도 생물공학의 동향(XII)
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    • pp.550-553
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    • 2003
  • $Hisx_6-GFPuv-Cyt$ c fusion protein을 E. coli 균주 JM109과 BL21 각각에서 발현시킨 결과 발현온도는 $30^{\circ}C$보다 $37^{\circ}C$에서, BL21보다 JM109에서의 발현량이 더 많았다. 그러나 JM109, $37^{\circ}C$에서 발현시킨 fusion protein의 $Ni^{2+}-IDA-agarose$ purification결과 약 45kDa 부근의 fusion protein의 density가 감소되었음을 SDS-PAGE analysis을 통해 알 수 있었다. 또한 western blotting analysis를 통해 이 impurity가 degraded fusion임을 확인 할 수 있었다. degraded fusion은 BL21 균주에서 발현시킨 fusion protein에서도 생성됨을 확인하였다. 모든 결과를 종합해 볼때 $Hisx_6-GFPuv-Cyt$ c fusion protein의 발현은 IM109, $37^{\circ}C$에서 더 많았지만, BL21, $37^{\circ}C$에서 expression시킨 fusion protein이 보다 안정하다고 판단 되어진다. 향후 fusion protein이 bioelectronic device에 적용되려면 degraded fusion protein의 생성을 줄여 activity를 유지하도록 안정한 형태로 발현되어 순수하게 분리정제 되어야 하겠다.

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Anti Inflammatory Effect of Low Level Laser Irradiation on the LPS-stimulated Murine Immunocytes

  • Jin, Dan;Lee, Jong-Young;Cho, Hyun-Chul;Kim, Soo-Ki
    • Molecular & Cellular Toxicology
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    • 제1권2호
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    • pp.124-129
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    • 2005
  • Pro-inflammatory cytokines, such as tumor necrosis factor $(TNF)-{\alpha}$, interleukin-12 (IL-12) and interleukin $(IL-1)-{\beta}$, play a key role in causing inflammatory diseases, which are rheumatoid arthritis, Crohn's disease and sepsis. Accumulating evidences suggest that low level laser irradiation (LLLI) may have an anti-inflammatory action. However, there are few data regarding down regulation of Th1 immune response by using the diod typed laser emitting device for human patients. As a fundamental step in order to address this issue, we investigated immunological impact of the low level laser irradiation (10 mw laser diode with a wavelength of 630 nm) on expression of pro-inflammatory cytokines in murine immunocytes (splenocytes and peritoneal macrophages) in vitro. The LLLI on lipopolysaccharide (LPS 100 ng/ml)-stimulated murine splenocytes and macrophages, clearly down regulated mRNA expression of $TNF-{\alpha}$ and IL-12 in dose-dependent manner. In addition, LLLI significantly inhibits the NO production in the LPS-stimulated murine macrophages. This data suggests that LLLI (wavelength of 630 nm) may exert an anti-inflammatory action via modulation of pro-inflammatory cytokine and NO production pathway.

Multidrug resistance of coagulase-negative staphylococci isolated from rescued wild animals

  • Rhim, Haerin;Kim, Hong-Cheul;Na, Ki-Jeong;Han, Jae-Ik
    • 한국동물위생학회지
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    • 제42권4호
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    • pp.251-255
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    • 2019
  • Wildlife is a bio-indicator of environmental pollution by antimicrobial resistant bacteria or genes, however, there is no information on antimicrobial resistance in wildlife-origin bacteria. This study aimed to investigate the normal microbiota of staphylococci and their antimicrobial resistance in wildlife that did not take any antimicrobials. After sampling and bacterial isolation/identification, antimicrobial resistance profiles were examined by broth microdilution test, Kirby-Bauer disc diffusion test and mecA genetargeted PCR. Of 90 isolates from wildlife, 83 were coagulase-negative staphylococci while only 7 were coagulase-positive staphylococci. Methicillin-resistance was found in 63 (70%) isolates and 35 of 90 (38.9%) isolates were multidrug-resistant staphylococci. When considering that all of the animals did not take any medication or contacted any medical device before the sampling, the results indicate significantly high prevalence of antimicrobial resistance in wild environments. Further study would be necessary to investigate the transmission route of antimicrobial resistance.

Recent Advancements in Technologies to Detect Enterohaemorrhagic Escherichia coli Shiga Toxins

  • Jeongtae Kim;Jun Bong Lee;Jaewon Park;Chiwan Koo;Moo-Seung Lee
    • Journal of Microbiology and Biotechnology
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    • 제33권5호
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    • pp.559-573
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    • 2023
  • Shiga toxin (Stxs)-producing enterohaemorrhagic Escherichia coli (EHEC) and Shigella dysenteriae serotype 1 are major causative agents of severe bloody diarrhea (known as hemorrhagic colitis) and hemolytic uremic syndrome (HUS) associated with extraintestinal complications such as acute renal failure and neurologic impairment in infected patients under 9 years of age. Extreme nephrotoxicity of Stxs in HUS patients is associated with severe outcomes, highlighting the need to develop technologies to detect low levels of the toxin in environmental or food samples. Currently, the conventional polymerase chain reaction (PCR) or immunoassay is the most broadly used assay to detect the toxin. However, these assays are laborious, time-consuming, and costly. More recently, numerous studies have described novel, highly sensitive, and portable methods for detecting Stxs from EHEC. To contextualize newly emerging Stxs detection methods, we briefly explain the basic principles of these methods, including lateral flow assays, optical detection, and electrical detection. We subsequently describe existing and newly emerging rapid detection technologies to identify and measure Stxs.

Radiofrequency에 노출된 Chlamydomonas sp.의 mycosporine-like amino acids 생합성 유전자 발현 (Expression of Mycosporine-like Amino Acids Biosynthetic Genes in the Chlamydomonas sp. Exposed to Radiofrequency)

  • 황진익;모상현;장만;이건섭;이주연;김동균;이택견
    • 한국산학기술학회논문지
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    • 제14권8호
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    • pp.4086-4092
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    • 2013
  • Mycosporine-like 아미노산(MAAs)은 UV 흡수물질이며, 다양한 해양생물들은 MAAs의 합성과 축적을 통하여 환경 자외선의 직 간접적인 영향을 감소시키는 기능을 진화시켜 왔다. 이 연구에서 우리는 radiofrequency(RF) 발생장치를 제작하였고, 이를 미세조류 배양에 적용하였다. $0.35{\pm}0.05$ mHz의 RF를 Chlamydomonas sp. 배양기에 공급하였고, 정해진 시간(0, 0.5, 1 및 2 시간)에 시료를 채취하였다. MAAs 생합성 관련 유전자인 dehydroquinate synthase homolog (DHQS-like)와 nonribosomal peptide synthetase homolog (NRPS-like) 유전자를 Chlamydomonas sp.로부터 클로닝하였고, RF 노출에 대한 유전자 발현을 qRT-PCR을 이용하여 분석하였다. 연구결과 RF에 노출된 Chlamydomonas sp.의 DHQS-like와 NRPS-like 유전자의 발현은 노출 1시간에 각각 1.46 배 및 1.19 배 증가하였다. 이러한 결과는 DHQS-like와 NRPS-like 유전자가 Chlaydomonas sp.의 MAAs 생합성을 진단할 수 있는 좋은 바이오마커 후보가 될 수 있음을 의미한다.

가토 하악골에서 신연 골형성술시 신연속도에 따른 TGF-$\beta$1, IGF-I, bFGF의 발현 (THE EXPRESSION OF TGF-$\beta$1, IGF-I, BFGF IN DISTRACTION OSTEOGENESIS ACCORDING TO DIFFERENT DISTRACTION RATES IN RABBIT'S MANDIBLE)

  • 신선아;지유진;송현철
    • Maxillofacial Plastic and Reconstructive Surgery
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    • 제27권3호
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    • pp.205-217
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    • 2005
  • Distraction osteogenesis is a technique of lengthening bone including soft tissue by gradual separation of surgically divided bone surfaces. Although the biomechanical, histological, and ultrastructural changes associated with distraction osteogenesis have been widely described, the molecular mechanisms governing the formation of new bone in distracted bone segments remain largely unclear. However, such information has significant clinical implications because it may enable targeted therapeutic manipulations designed to accelerate osseous regeneration. The purpose of this study was to evaluate the expression of TGF-$\beta$1, IGF-I and bFGF in distraction osteogenesis according to different distraction rates in a rabbit's mandible. When twenty-four adult rabbits underwent open osteotomy between the premolar and mental foramen, an external bilateral distraction device was applied. Latency was allowed for five days before distraction. Three different distraction rates were 0.7 mm/day (A, n=8), 1.4 mm/day (B, n=8) and 2.4 mm/day (C, n=8). The distraction device was activated with the same distraction rhythms of twice a day until 4.9 mm (A & B group) and 8.4 mm (C group) length gains was achieved. The animals were sacrificed at postoperative 3, 7, 14 and 28 days. The bony specimens were stained with H&E for histologic examination, and RT-PCR analysis was done for the identification of the expression of TGF-$\beta$1, IGF-I and bFGF. The results obtained from this study were as follows : The 0.7 mm/day and 1.4 mm/day distraction rate groups were shown to improve regenerative bone formation on radiographic and histologic examination. Also, TGF-$\beta$1, IGF-I and bFGF expression increased in the 0.7 mm/day and 1.4 mm/day distraction rate groups. But the 2.4 mm/day distraction rate group specimen was different with adjacent normal bone and hardly expressed of growth factors. These findings suggest that improved new bone formation in the 0.7 mm/day and 1.4 mm/day distraction rates is associated with enhanced expression of TGF-$\beta$1, IGF-I and bFGF by mechanical tension stress. Additionally, the 0.7 mm/day and 1.4 mm/day distraction rate groups were significantly different from the 2.4 mm/day distraction rate group in the expression of growth factors. According to the above results, it seems possible to apply a distraction rate of up to 1.4 mm/day a day in rabbit's mandible. And further studies are needed to evaluate growth factors of TGF-$\beta$1 and IGF-I, which are excellent in expression.

가토 하악골에서 신연 골형성술시 Osteonectin의 발현 (EXPRESSION OF OSTEONECTIN ON DISTRACTION OSTEOGENESIS IN THE RABBIT MANDIBLE)

  • 김동준;지유진;송현철
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • 제30권5호
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    • pp.391-399
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    • 2004
  • Distraction osteogenesis has been thought to be promising technique for replacing bone graft in maxilla and mandible. The purpose of this study was to investigate the expression of osteonectin on distraction osteogenesis. Sixteen rabbits were used for this experiment. Osteotomy was performed between premolar and mental foramen. On the experimental group, distraction device was connected to the respective bone segments. On the control group, bone segments were fixed using plate and screws after osteotomy. Distraction was carried out at the rate of 0.7mm per day to obtain a 4.9mm elongation on the experimental group. After 3 days, 7 days, 14 days, and 28 days two rabbits of each group were sacrificed. The results obtained from this study were as follow : Experimental group was observed that the gaps between the distracted bone edges were occupied by new bone. Expression of Osteonectin were detected throughout the experiment in both groups and Expression of Osteonectin were markedly increased during distraction and consolidation period in experimental group than control group. From these results, it could be stated that distraction was shown to improve and accelerate bone formation and mechanical stress like distraction has considerable effects on osteonectin.

오리사 바닥 깔짚자동살포장치 개발 및 실증 (Development and Field-evaluation of Automatic Spreader for Bedding Materials in Duck Houses)

  • 권경석;우제석;노재희;오상익;김종복;김중곤;양가영;장동화;최성민
    • 한국농공학회논문집
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    • 제63권1호
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    • pp.37-48
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    • 2021
  • The automatic-spreader of bedding materials was developed to reduce labor cost, and to achieve successful biosecurity in duck houses. Algorithm of the device was designed to realize a concept of the automatic unmanned operation including entire processes such as loading and spreading of the bedding materials. From the field measurement, the relationship between the expected water content reduction and weight of bedding materials per unit area according to the operation condition of the devices was induced. In the case of the measurement of particulate matters during the process of spreading works, the results of using both conventional manual-spreader and automatic-spreader were exceeded the allowable limit of inhalable and respirable dust, respectively. But, workers using automatic-spreader could be free from harmful aero-condition because they did not stay inside the facility during the spreading works. In addition, from the Duck hepatitis virus PCR test, influence on pulmonary tissue of the duck was not found. It could be expected that the development of the automatic-spreader of bedding materials for duck house can contribute to the advancement of duck breeding facilities.

바이오 시료의 적정온도 사이클 유지를 위한 채널형 히트싱크에 대한 실험적 연구 (Experimental Study on the Channel Type Heat Sink to Maintain Proper Temperature Cycle of Bio-Sample)

  • 황정규;박상희
    • 한국산업융합학회 논문집
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    • 제26권1호
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    • pp.183-191
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    • 2023
  • This study was conducted experimentally to investigate the surface temperature of the heat sink, the air temperature in the flow channel and the sample temperature by changing the channel number of channel type heat sink and the air flow rate when heating and cooling the bio sample. The target temperature of the sample was 15℃ or less as the minimum value and 82℃ or more as the maximum value. In this study, the channel number of the heat sink(N = 1, 2, 4, 5, 10) and the air flow rate(Q=25, 42, 54m3/min) were varied. The bio sample was replaced with water, and the volume of water is 4mL. The size of the heat sink is 80x73x150mm and the material is aluminum. When cooling the sample, the surface temperature, the air temperature and the sample temperature were highly dependent on the number of channels and the flow rate. However, when the sample is heated, the surface temperature, air temperature and sample temperature do not depend on the number of channels and the flow rate. It was found that the conditions for satisfying the minimum temperature of 15℃ or less when cooling the sample were the number of channels N≥5 and the flow rate Q≥42m3/min. When heating the sample, the conditions to satisfy the maximum temperature of 82℃ or more are the number of channels N≤5 and the air flow rate Q≤42m3/min.