• 제목/요약/키워드: two-probe method

검색결과 269건 처리시간 0.034초

Cloning and Expression of a Yeast Cell Wall Hydrolase Gene (ycl) from Alkalophilic Bacillus alcalophilus subsp. YB380

  • Ohk, Seung-Ho;Yeo, Ik-Hyun;Yu, Yun-Jung;Kim, Byong-Ki;Bai, Dong-Hoon
    • Journal of Microbiology and Biotechnology
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    • 제11권3호
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    • pp.508-514
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    • 2001
  • A stuructural gene (ycl) encoding novel yeast cell wall hydrolase, YCL, was cloned from alkalophilic Bacillus alcalophilus subsp. YB380 by PCR, and transformed into E. coli JM83. Based on the N-terminal and internal amino acid sequences of the enzyme, primers were designed for PCr. The positive clone that harbors 1.8 kb of the yeast cell wall hydrolase gene was selected by the colony hybridization method with a PCR fragment as a probe. According to the computer analysis, this gene contained a 400-base-paired N-terminal domain of the enzyme. Based on nucletide homology of the cloned gene, a 850 bp fragment was amplified and the C-terminal domain of the enzyme was sequenced. With a combination of the two sequences, a full nucleotide sequence for YCL was obtained. This gene, ycl, consisted of 1,297 nucleotides with 27 nucleotides with 27 amino acids of signal sequence, 83 redundant amino acids of prosequence, and 265 amino acids of the mature protein. This gene was then cloned into the pJH27 shuttle vector and transformed into the Bacillus subtilis DB104 to express the enzyme. It was confirmed that the expressed cell wall hydrolase that was produced by Bacillus subtilis DB104 was the same as that of the donor strain, by Western blot using polyclonal antibody (IgY) prepared from White Leghorn hen. Purified yeast cell wall hydrolase and expressed recombinant protein showed a single band at the same position in the Western blot analysis.

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유전자형에 따른 Streptococcus mutans의 subtyping: Southern blot RFLP와 AP-PCR을 이용한 비교 (EVALUATING TWO METHODS FOR FINGERPRINTING GENOMES FOR STREPTOCOCCUS MUTANS IN CHILDREN : A COMPARISON WITH AP-PCR AND SOUTHERN BLOT RFLP)

  • 정태성;김신
    • 대한소아치과학회지
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    • 제25권2호
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    • pp.292-303
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    • 1998
  • The arbitrary primer polymerase chain reaction(AP-PCR) and Southern blot restriction fragment length polymorphism(RFLP) were used to genotype the cariogenic pathogen S. mutans in children. Following the morphologic chracteristics of colony on selective medium for S. mutans, total genomic DNA from 155 strains was extracted by conventional methods. Among 155 strains, 143 strains (92.3%) were confirmed S. mutans by PCR with dexA gene and 114 strains were used in this study. Three random sequence 10-base oligonucleotide primers were chosen for AP-PCR. The amplified DNA products were separated electrophoretically in a 2% agarose gel containing ethidium bromide and the banding patterns were compared among different strains. For RFLP analysis, DNA was digested with EcoRI and BamHI, separated on a 0.7 % agarose gel and transferred to a nylon membrane. The membrane was probed with a previously characterised 1.6 kilobases (kb) DNA fragment cloned from gtf B gene of S. mutans. The probe was labeled with isotope[$^{32}P-{\alpha}CTP$], and hybridized fragments were detected with intensifying screen. AP-PCR produced 4-8 DNA bands in the 0.25-10 kb regions and distinguished 9, 10 or 12 genotypes, depending on the specific primer used. Southern blot RFLP analysis revealed 2 hybridization patterns consisting of 1 DNA fragments 450, 500 bp. These results indicate that AP-PCR is more discriminative method for genotyping of S. mutans.

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광섬유 Sagnac 간섭계를 이용한 초음파의 비접촉식 감지 (Non-contact Detection of Ultrasonic Waves Using Fiber Optic Sagnac Interferometer)

  • 이정주;장태성;이승석;김영길;권일범;이왕주
    • 대한기계학회논문집A
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    • 제25권9호
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    • pp.1400-1409
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    • 2001
  • This paper describes a fiber optic sensor suitable for non-contact detection of ultrasonic waves. This sensor is based on a fiber optic Sagnac interferometer. Quadrature phase bias between two interfering laser beams in Sagnac loop is introduced by a polarization controller. A stable quadrature phase bias can be confirmed by observing the interferometer output versus phase bias. This method eliminates a digital signal processing for detection of ultrasonic waves using Sagnac interferometer. Interference intensity is affected by the frequency of ultrasonic waves and the time delay of Sagnac loop. Collimator is attached to the end of the probing fiber to focus the light beam onto the specimen surface and to collect the reflected light back into the fiber probe. Ultrasonic waves produced by conventional ultrasonic transducers are detected. This fiber optic sensor based on Sagnac interferometer is very effective for detection of small displacement with high frequency such as ultrasonic waves used in conventional non-destructive testing.

Quenching Effect in an Optical Fiber Type Small Size Dosimeter Irradiated with 290 MeV·u-1 Carbon Ions

  • Hirata, Yuho;Watanabe, Kenichi;Uritani, Akira;Yamazaki, Atsushi;Koba, Yusuke;Matsufuji, Naruhiro
    • Journal of Radiation Protection and Research
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    • 제41권3호
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    • pp.222-228
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    • 2016
  • Background: We are developing a small size dosimeter for dose estimation in particle therapies. The developed dosimeter is an optical fiber based dosimeter mounting an radiation induced luminescence material, such as an OSL or a scintillator, at a tip. These materials generally suffer from the quenching effect under high LET particle irradiation. Materials and Methods: We fabricated two types of the small size dosimeters. They used an OSL material Eu:BaFBr and a BGO scintillator. Carbon ions were irradiated into the fabricated dosimeters at Heavy Ion Medical Accelerator in Chiba (HIMAC). The small size dosimeters were set behind the water equivalent acrylic phantom. Bragg peak was observed by changing the phantom thickness. An ion chamber was also placed near the small size dosimeters as a reference. Results and Discussion: Eu:BaFBr and BGO dosimeters showed a Bragg peak at the same thickness as the ion chamber. Under high LET particle irradiation, the response of the luminescence-based small size dosimeters deteriorated compared with that of the ion chamber due to the quenching effect. We confirmed the luminescence efficiency of Eu:BaFBr and BGO decrease with the LET. The reduction coefficient of luminescence efficiency was different between the BGO and the Eu:BaFBr. The LET can be determined from the luminescence ratio between Eu:BaFBr and BGO, and the dosimeter response can be corrected. Conclusion: We evaluated the LET dependence of the luminescence efficiency of the BGO and Eu:BaFBr as the quenching effect. We propose and discuss the correction of the quenching effect using the signal intensity ratio of the both materials. Although the correction precision is not sufficient, feasibility of the proposed correction method is proved through basic experiments.

감광성 CNT paste에 대한 저에너지 Ball Milling 처리 효과 (Effect of Ball Milling on Photosensitive Carbon Nanotube Pastes and Their Field Emission Properties)

  • 장은수;이한성;이내성
    • 한국전기전자재료학회:학술대회논문집
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    • 한국전기전자재료학회 2008년도 하계학술대회 논문집 Vol.9
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    • pp.154-154
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    • 2008
  • Although the screen printing technology using photosensitive carbon nanotube (CNT) paste has many advantages such as low cost, simple process, uniform emission, and capability of mass production, the CNT paste needs to be improved further in CNT dispersion, printability, adhesion, electrical conductivity, population of CNT emitters, etc. Ball milling has been frequently employed to prepare the CNT paste as ball milling can mix its ingredients very well and easily cut the long, entangled CNTs. This study carried out a parametric approach to fabricating the CNT paste in terms of low-energy ball milling and a paste composition. Field emission properties of the CNT paste was characterized with CNT dispersion and electrical conductivity which were measured by a UV-Vis spectrophotometer and a 4-point probe method, respectively. Main variables in formulating the CNT paste include a length of milling time, and amounts of CNTs and conductive inorganic fillers. In particular, we varied not only the contents of conductive fillers but also used two different sizes of filler particles of ${\mu}m$ and nm ranges. Among many variations of conductive fillers, the best field emission characteristics occurred at the 5 wt% fillers with the mixing ratio of 3:1 for ${\mu}m$-and nm-sizes. The amount and size of fillers has a great effect on the morphology, processing stability, and field emission characteristics of CNT emitter dots. The addition a small amount of nm-size fillers considerably improved the field emission characteristics of the photosensitive CNT paste.

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전립선암의 온열치료를 위한 초음파변환기 개발에 관한 연구 (A study for implementation of ultrasonic transducer in the prostate cancer hyperthermia)

  • 박문규;노시철;박재현;최흥호
    • 센서학회지
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    • 제18권5호
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    • pp.377-384
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    • 2009
  • The ultrasonic hyperthermia for oncology has been developed and studied. The HIFU(high intensity focused ultrasound) is the most recent method to treat the tumor by using ultrasound. In this study, an insertion-type transducer for treating a prostate cancer, which can focus the ultrasonic beam mechanically and electrically, was designed and developed. The developed transducer was composed of three arrays, and each array has 32 elements. For the purpose of the mechanical focusing, both side arrays are slanted to the center array by $15^{\circ}$. With this structure, NFL(near field length) was set up as 30 mm. The PZT-4 and two matching layers were used, and the backing layer was excepted to prevent energy losses. The acoustic field analysis and the heating test were performed to evaluate the performance of developed transducer. The shape of an acoustic field, peak pressure, and acoustic pressure distribution were compared with numerical simulation. The NFL was 32 mm, the beam width was 5 mm, focal area was $40\;mm^2$, and peak pressure was 5.5 MPa. With heating by using developed transducer, the temperature increased up to $33^{\circ}C$ at focal zone. As a result of this study, the usefulness of suggested transducer for prostate cancer hyperthermia was confirmed by the acoustic field analysis and the heating test with TMM(tissue mimicking) phantom.

Application of black phosphorus nanodots to live cell imaging

  • Shin, Yong Cheol;Song, Su-Jin;Lee, Yu Bin;Kang, Moon Sung;Lee, Hyun Uk;Oh, Jin-Woo;Han, Dong-Wook
    • 생체재료학회지
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    • 제22권4호
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    • pp.352-359
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    • 2018
  • Background: Black phosphorus (BP) has emerged as a novel class of nanomaterials owing to its unique optical and electronic properties. BP, a two-dimensional (2D) nanomaterial, is a structure where phosphorenes are stacked together in layers by van der Waals interactions. However, although BP nanodots have many advantages, their biosafety and biological effect have not yet been elucidated as compared to the other nanomaterials. Therefore, it is particularly important to assess the cytotoxicity of BP nanodots for exploring their potentials as novel biomaterials. Methods: BP nanodots were prepared by exfoliation with a modified ultrasonication-assisted solution method. The physicochemical properties of BP nanodots were characterized by transmission electron microscopy, dynamic light scattering, Raman spectroscopy, and X-ray diffractometry. In addition, the cytotoxicity of BP nanodots against C2C12 myoblasts was evaluated. Moreover, their cell imaging potential was investigated. Results: Herein, we concentrated on evaluating the cytotoxicity of BP nanodots and investigating their cell imaging potential. It was revealed that the BP nanodots were cytocompatible at a low concentration, although the cell viability was decreased with increasing BP nanodot concentration. Furthermore, our results demonstrated that the cells took up the BP nanodots, and the BP nanodots exhibited green fluorescence. Conclusions: In conclusion, our findings suggest that the BP nanodots have suitable biocompatibility, and are promising candidates as fluorescence probes for biomedical imaging applications.

습식 식각을 이용한 MWCNT-PMDS 변형율 센서 전극 생성에 관한 연구 (Electrode Fabrication of MWCNT-PDMS Strain Sensors by Wet-etching)

  • 정라희;황희윤
    • Composites Research
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    • 제34권6호
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    • pp.387-393
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    • 2021
  • 본 논문에서는 습식 식각으로 제작된 구리 전극을 가진 다중벽 탄소나노튜브 PDMS 복합재료 변형율 센서의 전기적 특성을 고찰하였다. MWCNT의 질량분율에 따라 MWCNT-PDMS 변형율 센서를 제작한 후, 전극 부착 표면을 습식 식각한 후 은-에폭시 전도성 접착제를 이용하여 구리 박판을 부착하였다. 2-프로브 방법으로 변형율 센서의 전기 전도성을 측정한 결과, 초기 저항은 MWCNT 함량과 식각 시간에 반비례하였지만 30% 변형율에 대한 저항 변화율은 MWCNT 함량과식각 시간에 비례하였다. 100회 반복 하중 시험 후 저항 변형율 감소는 MWCNT 함량이 증가할수록 식각 시간이 짧아질수록 상대적으로 작게 나타났다. 이는 식각에 의해 MWCNT-PDMS 변형율 센서의 초기 저항 감소에 기인한 것으로 판단된다.

Ag층을 이용한 Sn과 In의 무 플럭스 접합 (Fluxless Bonding Method between Sn and In Bumps Using Ag Capping Layer)

  • 이승현;김영호
    • 마이크로전자및패키징학회지
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    • 제11권2호
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    • pp.23-28
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    • 2004
  • 본 실험에서는 Ag 층을 이용한 무 플럭스 접합 공정을 개발하였으며 Ag의 유무에 따른 효과를 관찰하기 위해 In ($10{\mu}m$)과 Sn ($10{\mu}m$)솔더 및 Ag (100 nm)/In과 Ag/Sn 솔더를 thermal evaporation 방법으로 하부 금속층 위에 형성하였다. 접합부의 접촉저항과 전단 하중을 측정하기 위해 쿠폰시편을 제조하였으며 이리한 쿠폰시편은 $130^{\circ}C$에서 0.8, 1.6, 3.2 MPa의 접합압력을 가하여 30초간 접합을 실시하였다. 전단하중과 4단자 저항측정법을 이용하여 접합부의 특성을 분석하였으며 주사전자현미경(Scanning Electron Microscope), EDS (Energy Dispersive Spectrometry)과 X-ray mapping을 통해 접합부를 관찰하였다. 전단하중 측정 결과 0.8 MPa에서는 In-Sn 솔더의 접합이 이루어지지 않았으며 접합압력이 증가해도 Ag/In-Ag/Sn 시편의 전단하중 측정값이 In-Sn 시편에 비해 높게 나타났다. 접합부의 저항감은 $2-4\;m{\Omega}$을 나타내었으며 접합압력이 증가할수록 In-Sn 혼합층이 더 많이 관찰되었다.

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Line probe assay를 이용한 신속한 rifampicin내성결핵 진단법의 임상적 유용성 (Clinical Usefulness of the Line Probe Assay for Rapid Detection of Rifampicin-resistant Tuberculosis)

  • 홍상범;임채만;이상도;고윤석;김우성;김동순;김원동;심태선
    • Tuberculosis and Respiratory Diseases
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    • 제50권3호
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    • pp.334-342
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    • 2001
  • 배 경 : rpoB 유전자 돌연변이는 rifampicin 내성결핵의 96-98%에서 발견된다. 실험실 검사에서는 rpoB 유전자가 다제내성결핵의 지표로서 빠른 진단에 사용될 수 있음이 보고되었으나 임상적 적용에 대해서는 아직 국내 및 외국에도 보고가 없는 실정이다. 연구 방법 : 서울중앙병원에서 1998 년 6월부터 2000년 7월 까지 LiPA법을 이용하여 rpoB유전자 돌연변이분석이 시행된 33명 환자에서 후향적으로 의무기록을 조사하였다. 환자의 임상상, 약제 감수성, 그리고 LiPA 검사 결과를 비교 분석하였다. 전통적인 약제감수성 결과를 표준으로 하여 LiPA 검사 결과와 비교하였으며 양 검사결과가 불일치하는 경우에 rpoB 유전자 염기서열분석을 시행하였다. 연구결과 : 평균 나이는 $42{\pm}19$세이고, 남녀비는 24 : 9 이었다. rpoB 유전자 검사 요청 시간으로부터 결과 보고까지 평균 시간은 $5.2{\pm}2.6$일 이었다. rpoB 유전자 검사 결과는 약제 감수성 결과보다 평균시간은 $56{\pm}35$일 빠르게 보고되었다. 감수성 결과가 얄려진 33명 중 28(85%)명에서는 유전자 검사와 동일한 결과를 보였고, 5명은(15%) 반대 결과를 보였다. 반대 결과를 보인 5명에서 염기서열 분석을 하였을 때, 3예에서 약물 감수성 결과가 오류였을 가능성이 있고, 나머지 2예는 LiPA 검사결과가 오류였을 가능성이 있다. 치료 약 선택에 도움을 준 경우가 28예(85%) 있었다. 결 론 : LiPA 방법을 이용한 rpoB 유전자 검사는 임상에서도 다수에서 다제내성을 신속하고 정확하게 진단할 수 있었고, 치료약 선택에 도움을 주었다. 하지만 현재의 시점에서 LiPA 방법이 기존의 도말 및 배양검사를 완전히 대치할 수는 없고 임상상, 도말 및 배양검사결과와 종합하여 보조적으로 사용하면 도움이 될 것으로 사료되었다.

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