• Title/Summary/Keyword: signal sequence

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Molecular Cloning and Characterization of Bovine CYP26A1 Promoter (소 CYP26A1 유전자 프로모터의 molecular cloning 및 특성)

  • Kwak, Inseok
    • 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.

A Study on the Inhibition of Skin Pigmentation by Lobaric Acid as Protease Activated Receptor-2 Antagonist (Protease Activated Receptor-2의 길항제로서 Lobaric Acid의 피부 색소침착 억제 효능 연구)

  • Goo, Jung Hyun;Lee, Ji Eun;Myung, Cheol Hwan;Park, Jong Il;Hwang, Jae Sung
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.41 no.3
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    • pp.243-252
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    • 2015
  • Melanosome, the pigment granule in melanocyte, determines the color of skin when it moves into the keratinocyte. Inhibition of melanosome transfer from melanocyte to keratinocyte results in skin depigmentation. Protease activated receptor-2 (PAR-2) is involved in signal transduction systems via cell membrane and increases the melasome transfer when it is activated by cleavage of their extracellular amino acid sequence by trypsin or by a peptide such as SLIGKV. Here, we showed that lobaric acid inhibited PAR-2 activation and affected the mobilization of $Ca2^+$. The uptake of fluorescent microspheres and isolated melanosomes from melan-a melanocytes to keratinocytes induced by SLIGKV were inhibited by lobaric acid. Also, confocal microscopy studies illustrated a decreased melanosome transfer to keratinocytes in melanocyte-keratinocyte co-culture system by lobaric acid. In addition, lobaric acid induced visible skin lightening effect in human skin tissue culture model, melanoderm$^{(R)}$. Our data suggest that lobaric acid could be an effective skin lightening agent that works via regulation of phagocytic activity of keratinocytes.

Cloning of a Novel vpr Gene Encoding a Minor Fibrinolytic Enzyme from Bacillus subtilis SJ4 and the Properties of Vpr

  • Yao, Zhuang;Meng, Yu;Le, Huong Giang;Lee, Se Jin;Jeon, Hye Sung;Yoo, Ji Yeon;Kim, Hyun-Jin;Kim, Jeong Hwan
    • Journal of Microbiology and Biotechnology
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    • v.30 no.11
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    • pp.1720-1728
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    • 2020
  • We have previously characterized AprESJ4, the major fibrinolytic enzyme from Bacillus subtilis SJ4 (Yao et al., 2019). During that study, we observed a 68 kDa protein with fibrinolytic activity. In this study, we cloned the gene (vprSJ4) encoding the 68 kDa protein, a mature Vpr and minor protease secreted by Bacillus species. vprSJ4 encodes a preproenzyme consisting of 810 amino acids (aa) including signal sequence (28 aa) and prosequence (132 aa). The mature enzyme (650 aa) has a predicted molecular weight of 68,467.35. Unlike Vprs from other B. subtilis strains, VprSJ4 has 4 additional amino acids (DEFA) at the C-terminus. vprSJ4 was overexpressed in Escherichia coli. PreproVprSJ4 was localized in inclusion bodies, and subjected to in vitro renaturation and purification by an affinity column. SDS-PAGE and western blot showed that autoprocessing of preproVprSJ4 occurred and 68 kDa and smaller proteins were produced. The optimum pH and temperature of the recombinant VprSJ4 were pH 7.0 and 40℃, respectively. Kinetic parameters of recombinant VprSJ4 were measured by using an artificial substrate, N-succinyl-ala-ala-pro-phe-p-nitroanilide. Coexpression of vprSJ4 and aprESJ4 using pHY300PLK increased the fibrinolytic activity a further 117% when compared with aprESJ4 single expression using the same vector in B. subtilis WB600.

Susceptibility Vessel Sign for the Detection of Hyperacute MCA Occlusion: Evaluation with Susceptibility-weighted MR Imaging

  • Lee, Sangmin;Cho, Soo Bueum;Choi, Dae Seob;Park, Sung Eun;Shin, Hwa Seon;Baek, Hye Jin;Choi, Ho Cheol;Kim, Ji-Eun;Choi, Hye Young;Park, Mi Jung
    • Investigative Magnetic Resonance Imaging
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    • v.20 no.2
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    • pp.105-113
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    • 2016
  • Purpose: Susceptibility vessel sign (SVS) on gradient echo image, which is caused by MR signal loss due to arterial thrombosis, has been reported in acute middle cerebral artery (MCA) infarction. However, the reported sensitivity and diagnostic accuracy of SVS have been variable. Susceptibility-weighted imaging (SWI) is a newly developed MR sequence. Recent studies have found that SWI may be useful in the field of cerebrovascular diseases, especially for detecting the presence of prominent veins, microbleeds and the SVS. The purpose of this study was to evaluate the diagnostic values of SWI for the detection of hyperacute MCA occlusion. Materials and Methods: Sixty-nine patients (37 males, 32 females; 46-89 years old [mean, 69.1]) with acute stroke involving the MCA territory underwent MR imaging within 6 hours after the symptom onset. MR examination included T2, FLAIR (fluid-attenuated inversion recovery), DWI, SWI, PWI (perfusion-weighted imaging), contrast-enhanced MR angiography (MRA) and contrast-enhanced T1. Of these patients, 28 patients also underwent digital subtraction angiography (DSA) within 2 hours after MR examination. Presence or absence of SVS on SWI was assessed without knowledge of clinical, DSA and other MR imaging findings. Results: On MRA or DSA, 34 patients (49.3%) showed MCA occlusion. Of these patients, SVS was detected in 30 (88.2%) on SWI. The sensitivity, specificity, positive predictive value, negative predictive value and diagnostic accuracy of SWI were 88.2%, 97.1%, 96.8%, 89.5% and 92.8%, respectively. Conclusion: SWI was sensitive, specific and accurate for the detection of hyperacute MCA occlusion.

Characterization of a Revertant that Restroes the Export of Ribose-Bnding Potein to the Priplasm in Echerichia coli (리보스 결합 단백질을 페리플라슴으로 수송하는 복귀변이주의 분석)

  • ;;Randall, Linda L.
    • Korean Journal of Microbiology
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    • v.26 no.4
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    • pp.283-290
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    • 1988
  • A spontaneous revertant of mutation rbsB103 that is ribose taxis-positive was characterized. This revertant was found to be export-competent in the export of ribose-binding protein shown by the disappearance of accumulated mutant precursor protein and the export of mature ribose-binding protein to the periplasm. The reversional change was shown to be in the region of risB gene that codes for the amino terminal portion of ribose-binding protein. Analysis by high-performance liquid chromatography of peptide patterns of ribose-binding proteins confirmed the relationship between the wild-type and the revertant proteins as shown for the mutant previously (Iida et al., 1985). When the processing rate of presursor proteins from the wild type and the revertant strain in vivo was compared by pulse-chase experiment, it was found that processing is less efficient than normal in the revertant. Purified mature proteins from both wild-type and revertant were subjected to amino acid sequencing. The results confirmed the amino acid changes deduced from the DNA sequencing and showed that processing of the revertant precursor occured in the correct position even though there are two different amino acids present in the signal sequence.

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A Prediction Search Algorithm by using Temporal and Spatial Motion Information from the Previous Frame (이전 프레임의 시공간 모션 정보에 의한 예측 탐색 알고리즘)

  • Kwak, Sung-Keun;Wee, Young-Cheul;Kimn, Ha-Jine
    • Journal of the Korea Computer Graphics Society
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    • v.9 no.3
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    • pp.23-29
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    • 2003
  • There is the temporal correlation of the video sequence between the motion vector of current block and the motion vector of the previous block. If we can obtain useful and enough information from the motion vector of the same coordinate block of the previous frame, the total number of search points used to find the motion vector of the current block may be reduced significantly. In this paper, we propose the block-matching motion estimation using an adaptive initial search point by the predicted motion information from the same block of the previous frame. And the first search point of the proposed algorithm is moved an initial point on the location of being possibility and the searching process after moving the first search point is processed according to the fast search pattern. Simulation results show that PSNR(Peak-to-Signal Noise Ratio) values are improved UP to the 1.05dB as depend on the image sequences and improved about 0.33~0.37dB on an average. Search times are reduced about 29~97% than the other fast search algorithms. Simulation results also show that the performance of the proposed scheme gives better subjective picture quality than the other fast search algorithms and is closer to that of the FS(Full Search) algorithm.

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Frequency Synchronization Algorithm for Improving Performance of OFDMA System in 3GPP LTE Downlink (3GPP LTE 하향링크 OFDMA 시스템의 수신 성능 향상을 위한 주파수 동기 알고리즘)

  • Lee, Dae-Hong;Im, Se-Bin;Roh, Hee-Jin;Choi, Hyung-Jin
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.34 no.1C
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    • pp.120-130
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    • 2009
  • In this paper, we propose a receiver structure for frequency synchronization in OFDMA (Orthogonal Frequency Division Multiple Access) system which is considered as 3GPP LTE(Long Term Evolution) downlink. In general, OFDMA frequency synchronization consists of two parts: coarse synchronization and fine synchronization. We consider P-SCH (Primary-Synchronization Channel) and CP (Cyclic Prefix) of OFDMA symbol for coarse synchronization and fine synchronization, respectively. The P-SCH signal has two remarkable disadvantages that it does not have sufficiently many sub-carriers and its differential correlation characteristic is not good due to ZC (Zadoff Chu) sequence-specific property. Hence, conventional frequency synchronization algorithms cannot obtain satisfactory performance gain. In this paper, we propose a modified differential correlation algorithm to improve performance of the coarse frequency synchronization. Also, we introduce an effective PLL (Phase Locked Loop) structure to guarantee stable performance of the fine frequency synchronization. Simulation results verify that the proposed algorithm has superior performance to the conventional algorithms and the 2nd-order PLL is effective to track the fine frequency offset even in high mobility.

Novel Radix-26 DF IFFT Processor with Low Computational Complexity (연산복잡도가 적은 radix-26 FFT 프로세서)

  • Cho, Kyung-Ju
    • The Journal of Korea Institute of Information, Electronics, and Communication Technology
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    • v.13 no.1
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    • pp.35-41
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    • 2020
  • Fast Fourier transform (FFT) processors have been widely used in various application such as communications, image, and biomedical signal processing. Especially, high-performance and low-power FFT processing is indispensable in OFDM-based communication systems. This paper presents a novel radix-26 FFT algorithm with low computational complexity and high hardware efficiency. Applying a 7-dimensional index mapping, the twiddle factor is decomposed and then radix-26 FFT algorithm is derived. The proposed algorithm has a simple twiddle factor sequence and a small number of complex multiplications, which can reduce the memory size for storing the twiddle factor. When the coefficient of twiddle factor is small, complex constant multipliers can be used efficiently instead of complex multipliers. Complex constant multipliers can be designed more efficiently using canonic signed digit (CSD) and common subexpression elimination (CSE) algorithm. An efficient complex constant multiplier design method for the twiddle factor multiplication used in the proposed radix-26 algorithm is proposed applying CSD and CSE algorithm. To evaluate performance of the previous and the proposed methods, 256-point single-path delay feedback (SDF) FFT is designed and synthesized into FPGA. The proposed algorithm uses about 10% less hardware than the previous algorithm.

Up-regulation of dynamin-2 gene expression in Ras-transformed cells (Ras에 의해 암화된 세포에서 dynamin-2의 발현 촉진)

  • Yoo, Ji-Yun
    • Journal of Life Science
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    • v.17 no.3 s.83
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    • pp.375-380
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    • 2007
  • Dynamin plays a key role in the scission event common to various types of endocytosis. It has been previously reported that the SH3 domain-mediated association of Grb2 with dynamin-2 was dominantly found in Ras transformed cells. However, whether this association results from the increased expression of dynamin-2 and Grb2 in Ras transformed cells or not is still unknown. So in this study we first analyzed the expression levels of dynamin-2 and Grb2 and found that the expression of dynamin-2 protein was dramatically increased in Ras-transformed NIH3T3 (NIH3T3(Ras)) cells. Furthermore competitive PCR data revealed that the mRNA transcripts for dynamin-2 were increased about 100-fold in NIH3T3(Ras) compared to those of NIH3T3 cells. However, the protein level and mRNA transcript of Grb2 were not changed in these two cells. We also examined promoter activity of dynamin-2 in NIH3T3(Ras) cells and suggest the existence of Ras-responsive sequence in promoter region -300 to -200.

Increased Production of an Alkaline Protease from Bacillus clausii I-52 by Chromosomal Integration (Bacillus clausii I-52의 Chromosomal Integration에 의한 Alkaline Protease의 생산성 향상)

  • Joo, Han-Seung;Park, Dong-Chul;Choi, Jang-Won
    • Journal of agriculture & life science
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    • v.46 no.1
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    • pp.163-176
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    • 2012
  • TTo increase productivity of a strong extracellular alkaline protease (BCAP), stable strains of Bacillus clausii I-52 carrying another copy of BCAP gene in the chromosome were developed. Integrative vector, pHPS9-fuBCAP carrying BCAP promoter, ribosome binding site, signal sequence and active protease gene was constructed and transferred into B. clausii I-52, and integration of the constructed plasmid into chromosome was identified by PCR. An investigation was carried out on BCAP production by B. clausii I-52 and transformant C5 showing the highest relative activity of alkaline protease using submerged fermentation. Maximum enzyme activity was produced when cells were grown under the submerged fermentation conditions at $37^{\circ}C$ for 48 h with an aeration rate of 1 vvm and agitation rate of 650 rpm in a optimized medium (soybean meal 2%, wheat flour 1%, sodium citrate 0.5%, $K_2HPO_4$ 0.4%, $Na_2HPO_4$ 0.1%, NaCl 0.4%, $MgSO_47H_2O$ 0.01%, $FeSO_47H_2O$ 0.05%, liquid maltose 2.5%, $Na_2CO_3$ 0.6%). A protease yield of approximately 134,670U/ml was achieved using an optimized media, which show an increase of approximately 1.6-fold compared to that of non-transformant (83,960 U/ml). When the stability of transformant C5 was examined, the integrated plasmid pHPS9-fuBCAP was detected in the transformant after cultivation for 8 days, suggesting that it maintained stably in the chromosomal DNA of transformant C5.