• Journal of Microbiology
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• v.45 no.2
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• pp.158-167
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• 2007

In this study, we have cloned a novel cDNA encoding for a papain-family cysteine protease from the Uni-ZAP XR cDNA library of the polychaete, Periserrula leucophryna. This gene was expressed in Escherichia coli using the T7 promoter system, and the protease was characterized after partial purification. First, the partial DNA fragment (498 bp) was amplified from the total RNA via RT-PCR using degenerated primers derived from the conserved region of cysteine protease. The full-length cDNA of cysteine protease (PLCP) was prepared via the screening of the Uni-ZAP XR cDNA library using the $^{32}P-labeled$ partial DNA fragment. As a result, the PLCP gene was determined to consist of a 2591 bp nucleotide sequence (CDS: 173-1024 bp) which encodes for a 283-amino acid polypeptide, which is itself composed of an 59-residue signal sequence, a 6-residue propeptide, a 218-residue mature protein, and a long 3'-noncoding region encompassing 1564 bp. The predicted molecular weights of the preproprotein and the mature protein were calculated as 31.8 kDa and 25 kDa, respectively. The results of sequence analysis and alignment revealed a significant degree of sequence similarity with other eukaryotic cysteine proteases, including the conserved catalytic triad of the $Cys^{90},\;His^{226},\;and\;Asn^{250}$ residues which characterize the C1 family of papain-like cysteine protease. The nucleotide and amino acid sequences of the novel gene were deposited into the GenBank database under the accession numbers, AY390282 and AAR27011, respectively. The results of Northern blot analysis revealed the 2.5 kb size of the transcript and ubiquitous expression throughout the entirety of the body, head, gut, and skin, which suggested that the PLCP may be grouped within the cathepsin F-like proteases. The region encoding for the mature form of the protease was then subcloned into the pT7-7 expression vector following PCR amplification using the designed primers, including the initiation and termination codons. The recombinant cysteine proteases were generated in a range of 6.3 % to 12.5 % of the total cell proteins in the E. coli BL21(DE3) strain for 8 transformants. The results of SDS-PAGE and Western blot analysis indicated that a cysteine protease of approximately 25 kDa (mature form) was generated. The optimal pH and temperature of the enzyme were determined to be approximately 9.5 and $35^{\circ}C$, respectively, thereby indicating that the cysteine protease is a member of the alkaline protease group. The evaluation of substrate specificity indicated that the purified protease was more active towards Arg-X or Lys-X and did not efficiently cleave the substrates with non-polar amino acids at the P1 site. The PLCP evidenced fibrinolytic activity on the plasminogen-free fibrin plate test.

• Korean Journal of Optics and Photonics
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• v.26 no.3
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• pp.155-161
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• 2015

We report the design and performance analysis of an off-axis three-mirror telescope as the fore optics for a new hyperspectral sensor aboard a small unmanned aerial vehicle (UAV), for low-altitude coastal remote sensing. The sensor needs to have at least 4 cm of spatial resolution at an operating altitude of 500 m, $4^{\circ}$ field of view (FOV), and a signal to noise ratio (SNR) of 100 at 660 nm. For these performance requirements, the sensor's optical design has an entrance pupil diameter of 70 mm and an F-ratio of 5.0. The fore optics is a three-mirror system, including aspheric primary and secondary mirrors. The optical performance is expected to reach $1/15{\lambda}$ in RMS wavefront error and 0.75 in MTF value at 660 nm. Considering the manufacturing and assembling phase, we determined the alignment compensation due to the tertiary mirror from the sensitivity, and derived the tilt-tolerance range to be 0.17 mrad. The off-axis three-mirror telescope, which has better performance than the fore optics of other hyperspectral sensors and is fitted for a small UAV, will contribute to ocean remote-sensing research.

• Journal of the Korean Vacuum Society
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• v.15 no.4
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• pp.331-337
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• 2006

Cavity-type beam position monitors were developed in collaboration with KEK to use for the future accelerators such as international linear collider (ILC) or x-ray free electron laser (XFEL) in PAL. BPM components such as BPM cavity, beam tubes, waveguides and feedthroughs were assembled by brazing at the same time to reduce mechanical errors during the fabrication. There are four screwed pins around outer rim of the cavity for the tuning of cavity frequency and x-y isolation. The resonance frequency of BPM is 6.422 GHz, the inner diameter of cavity is 53.822 mm, and the range of mechanical adjusting is $+ / - 250{\mu}m$. The x-y isolation was measured better than -40 dB after tuned. Test results of signal forms, x-y isolations, sensitivities are satisfied within requirements for the KEK ATF2 beam line.

• Journal of the korean academy of Pediatric Dentistry
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• v.33 no.3
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• pp.401-410
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• 2006

Dental caries results from localized demineralization of tooth enamel by acids of bacterial origin produced from the fermentation of dietary sugars. A group of related oral bacteria, collectively known as mutans streptococci, are implicated as the primary etiological agents of human caries. Within this group, Streptococcus mutans has been known as a causative agent for dental caries. As well as acid production yielding the demineralization of tooth enamel, adherence and colonization of S. mutans to the teeth are also important for their virulence Cell-surface fibrillar proteins, which mediate adherence to the salivary pellicle are virulence components of mutans streptococci, and primary candidates for a human caries vaccine. Here we report that the AgI/II gene from S. mutans GS-5 were cloned by PCR amplification of the bacterial chromosomal DNA and the integrity of cloned genes were confirmed by nucleotide sequencing. Sequence analyses showed the sequence alignment of 280 nucleotides between the cloned AgI/II and the reported sequence of S. mutans GS-5 showed the perfect match The cloned genes which signal nucleotide was truncated, were transferred into bacterial expression vector and the recombinant proteins were purified as His-tag fusion proteins In order to generate polyclonal antibodies against the recombinant proteins, AgI/II mr, some $100{\mu}g$ of the proteins was injected into mice three times. It can be used for an effective vaccine production to prevent dental caries caused by pathogenic S. mutans.

• Korean Journal of Clinical Laboratory Science
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• v.52 no.1
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• pp.1-17
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• 2020

Three-dimensional microscopic approaches in histopathology display multiplex properties that present puzzling questions for specimens as related to their comprehensive volumetric information. This information includes spatial distribution of molecules, three-dimensional co-localization, structural formation and whole data set that cannot be determined by two-dimensional section slides due to the inevitable loss of spatial information. Advancement of optical instruments such as two-photon microscopy and high performance objectives with motorized correction collars have narrowed the gap between optical theories and the actual reality of deep tissue imaging. However, the benefits gained by a prolonged working distance, two-photon laser and optimized beam alignment are inevitably diminished because of the light scattering phenomenon that is deeply related to the refractive index mismatch between each cellular component and the surrounding medium. From the first approaches with simple crude refractive index matching techniques to the recent cutting-edge integrated tissue clearing methods, an achievement of transparency without morphological denaturation and eradication of natural and fixation-induced nonspecific autofluorescence out of real signal are key factors to determine the perfection of tissue clearing and the immunofluorescent staining for high contrast images. When performing integrated laboratory workflow of tissue for processing frozen and formalin-fixed tissues, clear lipid-exchanged acrylamide-hybridized rigid imaging/immunostaining/in situ hybridization-compatible tissue hydrogel (CLARITY), an equipment-based tissue clearing method, is compatible with routine procedures in a histopathology laboratory.

• Journal of Life Science
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• v.26 no.1
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• pp.42-49
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• 2016

The retinoic acid (RA) plays an important role in the growth and development of many cells, and bioactive RA concentration is regulated by several enzymes, including CYP26A1. The expression of the CYP26A1 gene is regulated by RA, and the CYP26A1 gene is one of the candidates for RA-responsive genes. Although CYP26A1 genes are cloned from several animals, cloning of the CYP26A1 gene from cows has not been reported yet. The promoter region of CYP26A1 from cows was cloned by PCR and analyzed by sequence alignment with human and mouse CYP26A1. The RA-responsive element (RARE), DR-5 (ttggg), was located in this region and was perfectly conserved. The promoter region of bovine CYP26A1, which contains DR-5, was ligated to the luciferase reporter gene on transient transfection assays. The expression of CYP26A1-Luc promoter was activated by ATRA treatment in lung-derived mtCC cells. Co-transfection with RAR-α or -β with ATRA significantly activates the expression of CYP26A1-Luc promoter; however, it was less effective with either RAR-γ or RXR-γ. In addition, the endogenous gene expressions measured by Q-RT-PCR in mtCC cells were not significantly affected by ATRA treatment for 2 days; however, the expression of the endogenous CYP26A1 gene was diminished sharply at day 3 with ATRA treatment. In conclusion, the promoter region of bovine CYP26A1 contains conserved DR-5 RARE, which functions as a binding site for RAR-α or -β, and it is involved in the regulation of CYP26A1 gene expression and the control of RA signaling in mtCC cells.

• Korean Journal of Optics and Photonics
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• v.32 no.2
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• pp.49-54
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• 2021

In this paper, we design and construct the equipment to manufacture large-diameter optical fiber end caps, which are the core parts of high-power fiber lasers, and we fabricate large-diameter optical fiber end caps using the home-made equipment. This equipment consists of a CO2 laser as a fusion-splice heat source, a precision stage assembly for transferring the position of a large-diameter optical fiber and an end cap, and a vision system used for alignment when the fusion splice is interlocked with the stage assembly. The output of the laser source is interlocked with the stage assembly to control the output, and the equipment is manufactured to align the polarization axis of the large-diameter polarization-maintaining optical fiber with the vision system. Optical fiber end caps were manufactured by laser fusion splicing of a large-diameter polarization-maintaining optical fiber with a clad diameter of 400 ㎛ and an end cap of 10×5×2 ㎣ (W×D×H) using home-made equipment. Signal-light insertion loss, polarization extinction ratio, and beam quality M2 of the fabricated large-diameter optical fiber end caps were measured to be 0.6%, 16.7 dB, and 1.21, respectively.

• The Journal of the Acoustical Society of Korea
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• v.28 no.2
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• pp.112-118
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• 2009

This paper proposes a rapid detection algorithm of a salient region for music video browsing system, which can be applied to mobile device and digital video recorder (DVR). The input music video is decomposed into the music and video tracks. For the music track, the music highlight including musical chorus is detected based on structure analysis using energy-based peak position detection. Using the emotional models generated by SVM-AdaBoost learning algorithm, the music signal of the music videos is classified into one of the predefined emotional classes of the music automatically. For the video track, the face scene including the singer or actor/actress is detected based on a boosted cascade of simple features. Finally, the salient region is generated based on the alignment of boundaries of the music highlight and the visual face scene. First, the users select their favorite music videos from various music videos in the mobile devices or DVR with the information of a music video's emotion and thereafter they can browse the salient region with a length of 30-seconds using the proposed algorithm quickly. A mean opinion score (MOS) test with a database of 200 music videos is conducted to compare the detected salient region with the predefined manual part. The MOS test results show that the detected salient region using the proposed method performed much better than the predefined manual part without audiovisual processing.