• Title/Summary/Keyword: X-vector

Search Result 595, Processing Time 0.037 seconds

Molecular Cloning and Expression of the Novel Attacin-Like Antibacterial Protein Gene Isolated from the Bombyx mori (집누에로부터 새로운 attactin 유산 항세균성 펩타이드 유전자의 분리 및 발현)

  • 윤은영;김상현;강석우;진병래;김근영;김호락;한명세;강석권
    • Korean journal of applied entomology
    • /
    • v.36 no.4
    • /
    • pp.331-340
    • /
    • 1997
  • Hyalophora cecropia attacin-like antibacterial gene was isolated from Bombyx mori induced with nonpathogenic bacteria. It was expressed in Spodopfera frugiperda 9 (Sf9 cells using baculovirus expression vector system (BEVS), and examined its antibacterial activity. With a cDNA library constructed from fifthinstar B. mori injected with Escherichia coli(4 X IOhcellsllarva), differential screening was performed using naive and induced mRNA probes. BmInc6 clone was screened by partial nucleotide sequence and GenBank database analysis. A complete nucleotide sequence of Bmlnc6 cDNA was determined (GenBank, AF005384). Its insert size was 852 bp and had open reading frame that started translation at position 35 and stopped at 679. And its putative polyadenylational signal existed at 812 bp. The number of amino acid deduced from Bmlnc6 cDNA was 214 and hydropathy analysis showed that this peptide was hydrophilic. This peptide deduced by BmInc6 was named nuecin. When the nuecin gene was expressed in Sf9 cells using BEVS, about 950 bp of the transcripts was detected. In addition, SDS-PAGE analysis showed that the molecular weights of intracellular expressed protein and the mature protein secreted to culture media were approximately 23 and 20 kDa, respectively. The antibacterial activity of nuecin against E. coli and Bacillus subtilis was significantly high, demonstrating that nuecin had a wider antibacterial spectrum with gram negative and positive bacteria than attacin.

  • PDF

Spectrofluorometric Characteristics of the N-Terminal Domain of Riboflavin Synthase (아미노-말단 리보플라빈 생성효소 단백질의 형광 특성)

  • Kim, Ryu-Ryun;Yi, Jeong-Hwan;Nam, Ki-Seok;Ko, Kyung-Won;Lee, Chan-Yong
    • Korean Journal of Microbiology
    • /
    • v.47 no.1
    • /
    • pp.14-21
    • /
    • 2011
  • Riboflavin synthase catalyzes the formation of one molecule of each riboflavin and 5-amino-6-ribitylamino-2,4-pyrimidinedione by the transfer of a 4-carbon moiety between two molecules of the substrates, 6,7-dimetyl-8-ribityllumazine. The most remarkable feature is the sequence similarity between the N-terminal half (1-97) and the C-terminal half domain (99-213). To investigate the structure and fluorescent characteristics of the N-terminal half of riboflavin synthase (N-RS) in Escherichia coli, more than 10 mutant genes coding for the mutated N-terminal domain of riboflavin synthase were generated by polymerase chain reaction. The genes coding for the proteins were inserted into pQE vector designed for easy purification of protein by 6X-His tagging system, expressed, and the proteins were purified. Almost all mutated N-terminal domain of riboflavin synthases bind to 6,7-dimethyl-8-ribityllumazine and riboflavin as fluorescent ligands. However, N-RS C47D and N-RS ET66,67DQ mutant proteins show colorless, indicating that fluorescent ligands were dissociated during purification. In addition, most mutated proteins show low fluorescent intensity comparing to N-RS wild type, whereas N-RS C48S posses stronger fluorescent intensity than that of wild type protein. Based on this result, N-RS C48S can be used as the tool for high throughput screening system for searching for the compound with inhibitory effect for the riboflavin synthase.

A Study of the Liposome-Mediated Transgenic Chicken Production (리포좀을 이용한 형질전환 닭 생산에 대한 연구)

  • Byun S. J.;Park C.;Yang B. S.;Kim T. Y.;Sohn S. H.;Kim S. H.;Jeon I. S.
    • Korean Journal of Poultry Science
    • /
    • v.31 no.4
    • /
    • pp.293-298
    • /
    • 2004
  • Microinjection of DNA is a general method for generating transgenic animals, but the rate of transgenesis in chickens is very low. So it was carried out to investigate the efficiency of liposome-mediated gene transfer in stage one cell of chicken embryo with GFP expression vector. In order to determine efficiency and duration of the introduced foreign gene, it was microinjected DNA with liposome or naked DNA into the germinal disc of stage one cell or stage-X chicken embryo. Analysis of reporter gene expression in day-4 embryos showed that GFP expression was observed only in the liposome-mediate embryo groups and detectable up to day-8 embryos. The results suggest that stable integration of the introduced gene using liposome is a rare event. Nevertheless the liposome-mediated gene transfer may be a useful method to transfer a foreign gene into the stage one cell of chicken embryos.

Analysis of Surface Plasmon Resonance on Periodic Metal Hole Array by Diffraction Orders

  • Hwang, Jeong-U;Yun, Su-Jin;Gang, Sang-U;No, Sam-Gyu;Lee, Sang-Jun;Urbas, Augustine;Ku, Zahyun
    • Proceedings of the Korean Vacuum Society Conference
    • /
    • 2013.02a
    • /
    • pp.176-177
    • /
    • 2013
  • Surface plasmon polaritons (SPPs) have attracted the attention of scientists and engineers involved in a wide area of research, microscopy, diagnostics and sensing. SPPs are waves that propagate along the surface of a conductor, usually metals. These are essentially light waves that are trapped on the surface because of their interaction with the free electrons of conductor. In this interaction, the free electrons respond collectively by oscillating in resonance with the light wave. The resonant interaction between the surface charge oscillation and the electromagnetic field of the light constitutes the SPPs and gives rise to its unique properties. In this papers, we studied theoretical and experimental extraordinary transmittance (T) and reflectance (R) of 2 dimensional metal hole array (2D-MHA) on GaAs in consideration of the diffraction orders. The 2d-MHAs was fabricated using ultra-violet photolithography, electron-beam evaporation and standard lift-off process with pitches ranging from 1.8 to $3.2{\mu}m$ and diameter of half of pitch, and was deposited 5-nm thick layer of titanium (Ti) as an adhesion layer and 50-nm thick layer of gold (Au) on the semiinsulating GaAs substrate. We employed both the commercial software (CST Microwave Studio: Computer Simulation Technology GmbH, Darmstadt, Germany) based on a finite integration technique (FIT) and a rigorous coupled wave analysis (RCWA) to calculate transmittance and reflectance. The transmittance was measured at a normal incident, and the reflectance was measured at variable incident angle of range between $30^{\circ}{\sim}80^{\circ}$ with a Nicolet Fourier transmission infrared (FTIR) spectrometer with a KBr beam splitter and a MCT detector. For MHAs of pitch (P), the peaks ${\lambda}$ max in the normal incidence transmittance spectra can be indentified approximately from SP dispersion relation, that is frequency-dependent SP wave vector (ksp). Shown in Fig. 1 is the transmission of P=2.2 um sample at normal incidence. We attribute the observation to be a result of FTIR system may be able to collect the transmitted light with higher diffraction order than 0th order. This is confirmed by calculations: for the MHAs, diffraction efficiency in (0, 0) diffracted orders is lower than in the (${\pm}x$, ${\pm}y$) diffracted orders. To further investigate the result, we calculated the angular dependent transmission of P=2.2 um sample (Fig. 2). The incident angle varies from 30o to 70o with a 10o increment. We also found the splitting character on reflectance measurement. The splitting effect is considered a results of SPPs assisted diffraction process by oblique incidence.

  • PDF

Biochemical Characterization of Cysteine(-) Mutant Alanine Racemase from Bacillus pseudomycoides (Bacillus pseudomycoides로 부터 분리된 alanine racemase 유전자의 cysteine 치환 및 생화학적 특성)

  • Kang, Han-Chul;Yoon, Sang-Hong;Lee, Chang-Muk;Koo, Bon-Sung
    • Journal of Applied Biological Chemistry
    • /
    • v.53 no.4
    • /
    • pp.195-201
    • /
    • 2010
  • A gene encoding an alanine racemase in B. pseudomycoides was cloned and one (Cys316) or both of two cysteines (Cys316 and Cys365) was (were) substituted with alanine. The cysteine (-) alanine racemases were expressed in E. coli BL21 (DE3) using a pET-21 vector. The expressed enzymes were purified through affinity chromatography using 6xHis ligand. The purified enzymes all showed major one bands by SDS-PAGE analysis, corresponding to 46 kDa. The cysteine (-) alanine racemases as well as the wild type enzyme showed alanine racemase activities, indicating that the enzyme is an alanine racemase and the cysteines in the enzyme may not be involved in the catalysis and/or substrate binding. Thermal stabilities of Cys (-) alanine racemases decreased considerably and half-lives were 26 (wild type), 21 (C316A) and 18 min (C316-365A), respectively at $60^{\circ}C$ pH 8.0, suggesting that cysteine is considerably contributive to the thermal stability of the alanine racemase.

Tn5 insertions in the agrocin 84 plasmid the conjugal nature of pAgK84 and the locations of determinants for transfer and agrocin 84 production

  • 심재섭
    • The Microorganisms and Industry
    • /
    • v.12 no.2
    • /
    • pp.2-13
    • /
    • 1986
  • Agrobacterium tumefaciens의 연구는 유전공학 시대를 맞이하여 많은 연구자들의 커다란 주목을 끌고있다. 식물세포내에 외부 유전자를 도입시키는, 확실히 믿을 수 있는 vector로 등장된 때문이다. 원래 이세균은 식물 줄기나 뿌리에 암종을 유발시키므로서 암발성 원인 구명 연구로 흥미를 끌게 되었다. 연구결과는 암발생 예방및 치료에 목적을 둠은 덩연할 것이다. 많은 약제가 시험되었으나 별로 진전을 보지 못하던중 비 원인성인 Agrobacterium radiobacter, strain 84에 의한 생물학적 방제의 성공으로 유일한 방제법을 갖게되었다. 뒤이어 암종발생 기작도 밝혀졌다. Agrobacterium의 세계는 온통 유전공학 기술로 채워져 있다. 암종발생에서 방제원리에 이르기까지 수없이 먼 옛날부터 이미 익혀오던 DNA 조작기술이었던가\ulcorner 암종을 유발시키는 agrocin84 plasmid를 갖는 비병원성 Agrobacterium을 찾아 생물학적 방제법을 확립하였다. 그후 병원성 Agrobacterium은 이에 대하여 어떻게 살아남을 것인가\ulcorner 실로 놀라운 일이라 아니할 수 있을까\ulcorner 이 병원성 Agrobacterium은 비 병원성 Agrobacterium 속에 있는 agrocin 84 plasmid을 탈취하여 자신이 agrocin84를 생성분비하며 암종 유발을 계소하여 간다. 아니면 비병원성 Agrobacterium이 병원성 Agrobacterium에게 agrocin 84 plasmid를 넘겨주었을까\ulcorner 왜 넘겨주었을까\ulcorner 공존을 위하여서일까\ulcorner 우리의 유전공학 기술은 이것을 막아줄수 있을까\ulcorner 생물학적 방제의 재성공을 위하여 논제의 연구는 왜 필요했던가\ulcorner 그 전후를 여기에 서술해 본다.닭이며 또한 제한된 지면에서 충분히 고찰하기는 어렵다. 우리나라에서 자주 거론되는 백신 및 종류에 국한하여 그 문제점과 앞으로의 전망을 고찰해 보기로 한다.ocking electrode를 제작하여 복합고분자 전해질과의 계면저항을 측정하였다.nm (1.2921eV)는 acceptor-bound exciton 인 I1(AO,X) 이고, 964.6nm(1.2853eV)는 donor-acceptor pair(DAP) 발광, 1341.9nm (0.9239eV)는 self activated(SA)에 기인하는 광발광 봉우리로 고찰되었다.가 높을수록 방출전류가 시간에 따라 급격히 감소하였다. 각 duty비에서 방출전류의 양이 1/2로 감소하는 시점을 에미터의 수명으로 볼 때 duty비 대 에미터 수명관계를 구해 높은 duty비에서 전계방출을 시킴으로써 실제의 구동조건인 낮은 duty비에서의 수명을 단시간에 예측할 수 있었다. 단속적으로 일어난 것으로 생각된다.리 폐 관류는 정맥주입 방법에 비해 고농도의 cisplatin 투여로 인한 다른 장기에서의 농도 증가 없이 폐 조직에 약 50배 정도의 고농도 cisplatin을 투여할 수 있었으며, 또한 분리 폐 관류 시 cisplatin에 의한 직접적 폐 독성은 발견되지 않았다이 낮았으나 통계학적 의의는 없었다[10.0%(4/40) : 8.2%(20/244), p>0.05]. 결론: 비디오흉강경술에서 재발을 낮추기 위해 수술시 폐야 전체를 관찰하여 존재하는 폐기포를 놓치지 않는 것이 중요하며, 폐기포를 확인하지 못한 경우와 이차성 자연기흉에 대해서는 흉막유착술에 더 세심한 주의가 필요하다는 것을 확인하였다. 비디오흉강경수술은 통증이 적고, 입원기간이 짧고, 사회로의 복귀가 빠르며, 고위험군에 적용할 수 있고, 무엇보다도 미용상의 이점이 크다는 면에서 자연기흉에 대해 유용한 치료방법임에는

  • PDF

A Study on the Construction of Indoor Spatial Information using a Terrestrial LiDAR (지상라이다를 이용한 지하철 역사의 3D 실내공간정보 구축방안 연구)

  • Go, Jong Sik;Jeong, In Hun;Shin, Han Sup;Choi, Yun Soo;Cho, Seong Kil
    • Spatial Information Research
    • /
    • v.21 no.3
    • /
    • pp.89-101
    • /
    • 2013
  • Recently, importance of indoor space is on the rise, as larger and more complex buildings are taking place due to development of building technology. Accordingly, range of the target area of spatial information service is rapidly expanding from outdoor space to indoor space. Various demands for indoor spatial information are expected to be created in the future through development of high technologies such as IT Mobile and convergence with various area. Thus this research takes a look at available methods for building indoor spatial information and then builds high accuracy three-dimensional indoor spatial information using indoor high accuracy laser survey and 3D vector process technique. The accuracy of built 3D indoor model is evaluated by overlap analysis method refer to a digital map, and the result showed that it could guarantee its positional accuracy within 0.04m on the x-axis, 0.06m on the y-axis. This result could be used as a fundamental data for building indoor spatial data and for integrated use of indoor and outdoor spatial information.

Inhibitory Mechanism on NF-${\kappa}B$ Transactivation by Dexamethasone in Pulmonary Epithelial Cells (폐상피세포에서 Dexamethasone에 의한 NF-${\kappa}B$ Transactivation 억제기전에 관한 연구)

  • Lee, Kye-Young;Kim, Yoon-Seop;Ko, Mi-Hye;Park, Jae-Seok;Jee, Young-Koo;Kim, Keun-Youl;Kwak, Sahng-June
    • Tuberculosis and Respiratory Diseases
    • /
    • v.48 no.5
    • /
    • pp.682-698
    • /
    • 2000
  • Glucocorticoid receptor (GR) functions as a suppressor of inflammation by inhibiting the expression of many cytokine genes activated by NF-${\kappa}B$. The goal of this study is to investigate the mechanism by which GR repress NF-${\kappa}B$ activation in lung epithelial cells. We used A549 and BEAS-2B lung epithelia! cell lines. Using Ig$G{\kappa}$-NF-${\kappa}B$ luciferase reporter gene construct, we found that dexamethasone significantly suppressed TNF-$\alpha$-induced NF-${\kappa}B$ activation and the overexpression of GR showed dose-dependent reduction of TNF-$\alpha$-induced NF-${\kappa}B$ activity in both cell lines. However, DNA binding of NF-${\kappa}B$ induced by TNF-$\alpha$ in electromobility shift assay was not inhibited by dexamethasone. Super shift assay with anti-p65 antibody demonstrated the existence of p65 in NF-${\kappa}B$ complex induced by $\alpha$ Western blot showed that $I{\kappa}B{\alpha}$ degradation induced by TNF-$\alpha$ was not affected by dexamethasone and $I{\kappa}B{\kappa}$ was not induced by dexamethasone, neither. To evaluate p65 specific transactivation, we adopted co-transfection study of Gal4-p65TA1 or TA2 fusion protein expression system together with 5xGal4-luciferase vector. Co-transfection of GR with Gal4-p65TA1 or TA2 repressed luciferase activity profoundly to the level of 10-20% of p65TA1- or TA2-induced transcriptional activity. And this transrepressional effect was abolished by co-transfection of CBP of SRC-1 expression vectors. These results suggest that GR-mediated transrepression of NF-${\kappa}B$ in lung epithelial cells is through competing for binding to limiting amounts of transcriptional coactivators, CBP or SRC-1.

  • PDF

Organ Specific Expression of the nos-NPT II Gene in Transgenic Hybrid Poplar (형질 전환된 포플러에 대한 nos-NPT II 유전자의 기관별 발현 특성)

  • Chun, Young Woo;Klopfenstein, Ned B.
    • Journal of Korean Society of Forest Science
    • /
    • v.84 no.1
    • /
    • pp.77-86
    • /
    • 1995
  • To effectively modify tree function with genetic engineering, transgenes must be expressed at the proper level in the appropriate tissues at suitable developmental stages. Toward understanding the spatial and temporal expression of transgenes in woody plants, transgene expression was evaluated in three greenhouse-grown, transgenic lines of Populus alba ${\times}$ P. grandidentata hybrid clone 'Hansen'. All transgenic poplar lines possess constructs containing the bacterial nopaline synthase(nos) promoter linked to a neomycin phosphotransferase II(NPT II) selectable marker gene. In addition, each transgenic poplar line contains one of the following gene constructs : 1) a wound-inducible potato proteinase inhibitor II (pin2) promoter linked to a chloramphenicol acetyltransferase(CAT) reporter gene. 2) a nos promoter linked to a PIN2 structural gene : or 3) a Cauliflower Mosaic Virus 35s promoter linked to a PIN2 structural gene. Polymerase chain reaction(PCR) was used to verify the presence of foreign genes in the poplar genome. Enzyme-linked immunosorbent assays(ELISAs) were used to evaluate organ specific expression of the nos-NPT II construct. NPT II expression was detected in leaves, petioles, stems, and roots of transgenic poplar, thereby indicating that the nos promoter is potentially effective for general constitutive expression of transgenes. NPT expression varied among transgenic poplar lines and among organs for one transgenic line, Tr15. With Tr15, NPT II levels were highest in older leaves and petioles. These results indicate that screening of several transgenic lines may be required to identify lines with optimal transgene expression.

  • PDF

The Determinants of New Supply in the Seoul Office Market and their Dynamic Relationship (서울 오피스 신규 공급 결정요인과 동태적 관계분석)

  • Yang, Hye-Seon;Kang, Chang-Deok
    • Journal of Cadastre & Land InformatiX
    • /
    • v.47 no.2
    • /
    • pp.159-174
    • /
    • 2017
  • The long-term imbalances between supply and demand in office market can weaken urban growth since excessive supply of offices led to office market instability and excessive demand of offices weakens growth of urban industry. Recently, there have been a lot of new large-scale supplies, which increased volatility in Seoul office market. Nevertheless, new supply of Seoul office has not been fully examined. Given this, the focus of this article was on confirming the influences of profitability, replacement cost, and demand on new office supplies in Seoul. In examining those influences, another focus was on their relative influences over time. For these purposes, we analyzed quarterly data of Seoul office market between 2003 and 2015 using a vector error correction model (VECM). As a result, in terms of the influences on the current new supply, the impact of supply before the first quarter was negative, while that of office employment before the first quarter was positive. Also, that of interest rate before the second quarter was positive, while those of cap rate before the first quarter and cap rate before the second quarter were negative. Based on the findings, it is suggested that prediction models on Seoul offices need to be developed considering the influences of profitability, replacement cost, and demand on new office supplies in Seoul.