• Journal of Microbiology and Biotechnology
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• 제12권3호
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• pp.437-443
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• 2002

A number of soil and wastewater samples were collected from the vicinity of an effluent treatment plant for the chemical industry. Several microorganisms were screened fur their ability to decolorize the triphenylmethane group of dyes. As a result, a novel crystal violet dye-degrading strain LK-24 was isolated. Taxonomic identification including 16S rDNA sequencing and phylogenetic analysis indicated that the isolate had a $99.5\%$ homology in its 16S rDNA base sequence with Stenotrophomonas maltophilia. The triphenylmethane dye, crystal violet, was degraded extensively by growing cells of Stenotrophomonas maltophilia LK-24 in agitated liquid cultures, although their growth was strongly inhibited in the initial stage of incubation. This group of dyes is toxic, depending on the concentration used. The dye was significantly degraded at a relatively lower concentration, below $100{\mu}g\;ml^-1$, yet the growth of the cells was totally suppressed at a dye concentration of $250{\mu}g\;ml^-1$. The degradation products of crystal violet were identified as 4,4'-bis(dimethylamino)-benzophenone and ${\rho}$-dimethylaminophenol by Gas chromatography-Mass spectrometry. The 4,4'-bis(dimethylamino)-benzophenone was easily obtained in a reasonable yield, as it was not metabolized further by S. maltophilia LK-24; however, the ${\rho}$-dimethylaminophenol was not easily identifiable, as it was further metabolized.

• Journal of Microbiology and Biotechnology
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• 제25권9호
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• pp.1449-1459
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• 2015

A novel proteolytic enzyme with fibrinolytic activity, FSP3, was purified from the recently isolated Streptomyces sp. P3, which is a novel bacterial strain isolated from soil. FSP3 was purified to electrophoretic homogeneity by ammonium sulfate precipitation, anion exchange, and gel filtration. FSP3 is considered to be a single peptide chain with a molecular mass of 44 kDa. The maximum activity of the enzyme was observed at 50℃ and pH 6.5, and the enzyme was stable between pH 6 and 8 and below 40℃. In a fibrin plate assay, FSP3 showed more potent fibrinolytic activity than urokinase, which is a clinical thrombolytic agent acting as a plasminogen activitor. The activity was strongly inhibited by the serine protease inhibitor PMSF, indicating that it is a serine protease. Additionally, metal ions showed different effects on the activity. It was significantly suppressed by Mg²⁺ and Ca²⁺ and completely inhibited by Cu²⁺, but slightly enhanced by Fe²⁺. According to LC-MS/MS results, its partial amino acid sequences are significantly dissimilar from those of previously reported fibrinolytic enzymes. The sequence of a DNA fragment encoding FSP3 contained an open reading frame of 1287 base pairs encoding 428 amino acids. FSP3 is a bifunctional enzyme in nature. It hydrolyzes the fibrin directly and activates plasminogen, which may reduce the occurrence of side effects. These results suggest that FSP3 is a novel serine protease with potential applications in thrombolytic therapy.

• Reproductive and Developmental Biology
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• 제33권1호
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• pp.63-69
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• 2009

In this study we tried to construct a more efficient tetracycline-inducible gene expression system by replacing previous key elements with more advance ones. At the beginning, we substituted PGK (phophoglycerate kinase) promoter for CMV (cytomegalovirus) promoter to control "$rtTA2^sM2$" which has been known for high induction efficiency in response to tetracycline. With this modification, expression of the EGFP marker gene under the induction condition was significantly increased. Next, we replaced "TRE" fragment with a modified version named "TRE-tighf" which has been reported to have higher affinity and specificity to the transactivator by minor base change of the "TRE" DNA fragment sequence. Use of "TRE-tighf" instead of "TRE" resulted in more than 10 fold increment in terms of induction efficiency and significant decrement of background expression in non-inducible condition. By combining PGK promoter and "TRE-tight" fragment, we could upgrade previous tetracycline-inducible system to show more stringent turn on/off gene switch ability and stronger expression of the gene of our interest. Use of this newly developed system must be very helpful to the studies of gene expression, especially to the transgenic animal study in which non-controllable constitutive expression of the transgene has been one of the urgent problems to be solved.

• Computers and Concrete
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• 제12권2호
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• pp.211-228
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• 2013

The structure analyzed in this paper has particular building style and special structural system. It is a rigid-connected twin-tower skyscraper with asymmetrical distribution of stiffness and masses in two towers. Because of the different stiffness between the north and the south towers, the torsion seismic vibration is significant. In this paper, in order to study the seismic response of the structure under both frequent low-intensity earthquakes as well as rare earthquakes at the levels of intensity 7, the analysis model is built and analyzed with NosaCAD. NosaCAD is an nonlinear structure analysis software based on second-development of AutoCAD with ObjectARX. It has convenient modeling function, high computational efficiency and diversity post-processing functions. The deformations, forces and damages of the structure are investigated based on the analysis. According to the analysis, there is no damage on the structure under frequent earthquakes, and the structure has sufficient capacity and ductility to resist rare earthquakes. Therefore the structure can reach the goal of no damage under frequent earthquakes and no collapse under rare earthquakes. The deformation of the structure is below the limit in Chinese code. The time sequence and distribution of damages on tubes are reasonable, which can dissipate some dynamic energy. At last, according to forces, load-carrying capacity and damage of elements, there are some suggestions on increasing the reinforcement in the core tube at base and in stiffened stories.

• Journal of Microbiology and Biotechnology
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• 제25권12호
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• pp.1971-1976
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• 2015

Oceanobacillus kimchii is a member of the genus Oceanobacillus within the family Bacillaceae. Species of the Oceanobacillus possess moderate haloalkaliphilic features and originate from various alkali or salty environments. The haloalkaliphilic characteristics of Oceanobacillus advocate they may have possible uses in biotechnological and industrial applications, such as alkaline enzyme production and biodegradation. This study presents the draft genome sequence of O. kimchii X50^T and its annotation. Furthermore, comparative genomic analysis of O. kimchii X50^T was performed with two previously reported Oceanobacillus genome sequences. The 3,822,411 base-pair genome contains 3,792 protein-coding genes and 80 RNA genes with an average G+C content of 35.18 mol%. The strain carried 67 and 13 predicted genes annotated with transport system and osmoregulation, respectively, which support the tolerance phenotype of the strain in high-alkali and high-salt environments.

• 한국미생물·생명공학회지
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• 제46권2호
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• pp.145-153
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• 2018

Among the carotenoid biosynthesis genes, crtI gene encodes the phytoene desaturase (CrtI) enzyme, and phytoene desaturase convert phytoene to lycopene. Phytoene desaturase is involved in the dehydrogenation reaction, in which four single bonds in the phytoene are introduced into a double bond, eliminating eight hydrogen atoms in the process. Phytoene desaturase is one of the key regulating enzyme in carotenoid biosynthetic pathway of various carotenoid biosynthetic organisms. The crtI gene in genomic DNA of Paracoccus haeundaensis was amplified and cloned into a T-vector to analyze the nucleotide sequence. As a result, the crtI gene coding for phytoene desaturase from P. haeundaensis consists of 1,503 base pairs encoding 501 amino acids residues. An expression plasmid containing the crtI gene was constructed, and Escherichia coli cells containing this plasmid produced the recombinant protein of approximately 55 kDa, equivalent to the molecular weight of phytoene desaturase. The expressed protein in cell lysate showed enzymatic activity similar to phytoene desaturase. Phytoene and lycopene were analyzed by HPLC and measured at wavelength of 280 nm and 470 nm, respectively. The $K_m$ values for phytoene and NADPH were $11.1{\mu}M$ and $129.3{\mu}M$, respectively.

• 한국정보과학회논문지:정보통신
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• 제34권3호
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• pp.156-167
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• 2007

본 논문에서는 VBR(Variable-Bit-Rate) 트래픽의 비선형적이고 버스티한 특성을 모델화 한 GOP ARIMA(ARIMA for Group Of Pictures) 모델을 칼만 필터 알고리즘을 이용하여 실시간으로 예측하는 기법을 제안한다. 칼만 필터를 이용한 예측 기법은 GOP ARIMA의 상태공간 모델링 과정과 향후 N초 간의 트래픽을 예측하는 과정으로 구성된다. 실험을 위해 GOP의 크기가 각각 15인 세 가지 종류의 MPEG VBR 트래픽(뉴스, 드라마, 스포츠)을 제작하였고, 칼만 필터를 이용한 세 가지 종류의 트래픽의 예측 결과를 선형 예측법과 이중 지수 평활법을 이용해 예측한 결과와 비교해 예측 성능이 상대적으로 우수함을 확인할 수 있었다. 또한 예측값에 신뢰 구간을 설정하는 신뢰 구간 분석법을 통해 트래픽 관점에서 장면 변화를 예측하는 방법을 제시하였다. 본 논문의 칼만 필터 기반의 예측 알고리즘은 MPEG 기반 VBR 트래픽을 비롯한 기타 인터넷 트래픽을 실시간으로 예측하는 방법과 이를 이용해 인터넷 서버의 설계 및 자원 할당 정책 등을 위한 트래픽 엔지니어링 연구에 기여할 수 있을 것이다.

• 한국멀티미디어학회논문지
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• 제15권11호
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• pp.1284-1291
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• 2012

선박 객체 검출 기술은 입력된 비디오 및 영상 데이터에서 선박 객체가 존재하는 경우 선박의 위치를 검출하는 기술로서 입력 영상의 환경 변화와 잡음의 영향에 따라서 검출 정확도의 편차가 높다. 이런 문제점을 해결하기 위하여 본 논문에서는 배경 구축 기법과 형태학적 연산 기반의 다중 선박 객체 검출 기술을 제안한다. 제안하는 방법은 배경 제거 단계, 잡음 제거 단계, 객체 기준 위치 설정 단계, 객체 재구성 단계, 다중 객체 검출 단계 등 5단계를 거쳐서 선박을 검출한다. 다양한 변수를 고려한 15가지 실험 비디오를 대상으로 한 실험을 통해서 98.7%의 검출율을 나타내었으며, 환경 변화에 강인한 검출을 수행하는 것을 확인할 수 있었다. 제안하는 방법은 해상 관제와 선박 자동 운항 기술의 기반 기술로서 유용하게 사용될 수 있다.

• 한국자원식물학회지
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• 제23권6호
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• pp.526-534
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• 2010

Genomes of clusters of related eukaryotes are now being sequenced at an increasing rate. In this paper, we developed an accurate, low-cost method for annotation of gene prediction and exon-intron structure. The gene prediction was adapted for delta 1-pyrroline-5-carboxylate-synthetase (p5cs) gene from China wild-type of the halophytic Leymus chinensis (Trin.), naturally adapted to highly-alkali soils. Due to complex adaptive mechanisms in halophytes, more attentions are being paid on the regulatory elements of stress adaptation in halophytes. P5CS encodes delta 1-pyrroline-5-carboxylate-synthetase, a key regulatory enzyme involved in the biosynthesis of proline, that has direct correlation with proline accumulation in vivo and positive relationship with stress tolerance. Using analysis of reverse transcription-polymerase chain reaction (RT-PCR) and PCR, and direct sequencing, 1076 base pairs (bp) of cDNA in length and 2396 bp of genomic DNA in length were obtained from direct sequencing results. Through gene prediction and exon-intron structure verification, the full-length of cDNA sequence was divided into eight parts, with seven parts of intron insertion. The average lengths of determinated coding regions and non-coding regions were 154.17 bp and 188.57 bp, respectively. Nearly all splice sites displayed GT as the donor sites at the 5' end of intron region, and 71.43% displayed AG as the acceptor sites at the 3' end of intron region. We conclude that this method is a cost-effective way for obtaining an experimentally verified genome annotation.

• Asian Pacific Journal of Cancer Prevention
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• 제16권11호
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• pp.4589-4592
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• 2015

Background: Missense and frame-shift mutations within the dimer forming domain of the caspase 8 gene have been identified in several cancers. However, the genetic status of this region in precancerous lesions, like oral submucous fibrosis (OSMF), and well differentiated oral squamous cell carcinomas (OSCCs) in patients from southern region of India is not known, and hence the present study was designed to address this issue. Materials and Methods: Genomic DNA isolated from biopsy tissues of thirty one oral submucous fibrosis and twenty five OSCC samples were subjected to PCR amplification with intronic primers flanking exon 7 of the caspase 8 gene. The PCR amplicons were subsequently subjected to direct sequencing to elucidate the status of mutation. Results: Sequence analysis identified a frame-shift and a novel missense mutation in two out of twenty five OSCC samples. The frame-shift mutation was due to a two base pair deletion (c.1225_1226delTG), while the missense mutation was due to substitution of wild type cysteine residue with phenylalanine at codon 426 (C426F). The missense mutation, however, was found to be heterozygous as the wild type C426C codon was also present. None of the OSMF samples carried mutations. Conclusions: The identification of mutations in OSCC lesions but not OSMF suggests that dimer forming domain mutations in caspase 8 may be limited to malignant lesions. The absence of mutations in OSMF also suggests that the samples analyzed in the present study may not have acquired transforming potential. To the best of our knowledge this is the first study to have explored and identified frame-shift and novel missense mutations in OSCC tissue samples.