• Title/Summary/Keyword: transfer vector

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Rabbit Hemorrhagic Disease Virus Variant Recombinant VP60 Protein Induces Protective Immunogenicity

  • Yang, Dong-Kun;Kim, Ha-Hyun;Nah, Jin-Ju;Song, Jae-Young
    • Journal of Microbiology and Biotechnology
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    • v.25 no.11
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    • pp.1960-1965
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    • 2015
  • Rabbit hemorrhagic disease virus (RHDV) is highly contagious and often causes fatal disease that affects both wild and domestic rabbits of the species Oryctolagus cuniculus. A highly pathogenic RHDV variant (RHDVa) has been circulation in the Korean rabbit population since 2007 and has a devastating effect on the rabbit industry in Korea. A highly pathogenic RHDVa was isolated from naturally infected rabbits, and the gene encoding the VP60 protein was cloned into a baculovirus transfer vector and expressed in insect cells. The hemagglutination titer of the Sf-9 cell lysate infected with recombinant VP60 baculovirus was 131,072 units/50 μl and of the supernatant 4,096 units/50 μl. Guinea pigs immunized twice intramuscularly with a trial inactivated RHDVa vaccine containing recombinant VP60 contained 2,152 hemagglutination inhibition (HI) geometric mean titers. The 8-week-old white rabbits inoculated with one vaccine dose were challenged with a lethal RHDVa 21 days later and showed 100% survival rates. The recombinant VP60 protein expressed in a baculovirus system induced high HI titers in guinea pigs and rendered complete protection, which led to the development of a novel inactivated RHDVa vaccine.

Cloning and Expression of $\beta$-Xylosidase Gene from Alkali-tolerant Bacillus sp. YA-14 in Escherichia coli (알카리 내성 Bacillus sp. YA-14의 $\beta$-Xylosidase 유전자의 Cloning 및 대장균에의 발현)

  • 박덕철;김진만;정용준;공인수;배동훈;유주현
    • Microbiology and Biotechnology Letters
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    • v.17 no.6
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    • pp.574-579
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    • 1989
  • Chromosomal DNA fragments of Bacillus sp. YA-14, isolated from soil as a potent $\beta$-xylosidase producing bacterium, were ligated to a vector plasmid pBR322 and used to transfer Escherichia coli HB101 cells. The recombinant plasmid pYXL22 was found to enable the transformants to produce $\beta$-xylosidase. pYXL22 was found to contain the 7.0 kb HindIII DNA fragment originated from the Bacillus sp. YA-14 chromosomal DNA by Southern hybridization. The optimum temperature for the reaction of $\beta$-xylosidase produced by E. coli HB101 (pYXL22) was appeared at 3$0^{\circ}C$. The enzyme was maintained stably up to 4$0^{\circ}C$ when stored 1hr at 4$0^{\circ}C$. The $\beta$-xylosidase was repressed completely by 0.4% (w/v) glucose concentration in E. coli HB101 (pYXL22). The optimum concentration of xylose for the $\beta$-xylosidase production in Bacillus sp. YA-14 was 0.2% (w/v).

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Construction and Transformation of an Endogenous Plasmid pBL1-free Brevibacterium lactofermentum (내재형 Plasmid pBL1이 제거된 Brevibacterium lactofermentum 개발과 형질전환)

  • 이규남;민본홍;윤기홍
    • Microbiology and Biotechnology Letters
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    • v.23 no.2
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    • pp.164-169
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    • 1995
  • An endogenous cryptic plasmid, pBL1, which has been used to construct plasmid vectors for coryneform bacteria producing amino acids, was eliminated from Brevibacterium lactofermentum. The pBL1 was partially digested with Sau3AI and the resulting DNA fragments were subcloned into a suicide vector pEM1 which contains a kanamycin-resistant (km$^{r}$) gene. KM$^{r}$ B. lactofermentum transconjugants were obtained by conjugal transfer of the pEM1 derivatives containing pBL1 DNA fragments from Escherichia coli into B. lactofermentum. A km$^{r}$ transconjugant was analyzed to contain a plasmid pEB14, which occurred in vivo by homologous recombination between pBL1 and the conjugal-transferred plasmid. The pEB14 including the pEM1-derived km$^{r}$ gene was found to be lost concomitantly with km$^{r}$ phenotype, resulting in the construction of a pBL1-free strain of B lactofermentum. Based on transformation efficiencies and plasmid stability, the resultant pBL1- free strain is more useful than wild strain as a host cell for genetic manipulation. It could be concluded that foreign plasmid DNAs are efficiently isolated and analyzed from the pBL1-free strain because of the absence of endogenous pBL1 plasmid.

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A Gammaherpesvirus Establishes Persistent Infection in Neuroblastoma Cells

  • Cho, Hye-Jeong;Song, Moon Jung
    • Molecules and Cells
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    • v.37 no.7
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    • pp.518-525
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    • 2014
  • Gammaherpesvirus (${\gamma}HV$) infection of the central nervous system (CNS) has been implicated in diverse neurological diseases, and murine ${\gamma}HV$-68 (MHV-68) is known to persist in the brain after cerebral infection. The underlying molecular mechanisms of persistency of virus in the brain are poorly understood. Here, we characterized a unique pattern of MHV-68 persistent infection in neuroblastoma cells. On infection with MHV-68, both murine and human neuroblastoma cells expressed viral lytic proteins and produced virions. However, the infected cells survived productive infection and could be cultured for multiple passages without affecting their cellular growth. Latent infection as well as productive replication was established in these prolonged cultures, and lytic replication was further increased by treatment with lytic inducers. Our results provide a novel system to study persistent infection of ${\gamma}HVs$ in vitro following de novo infection and suggest application of MHV-68 as a potential gene transfer vector to the brain.

Object Retrieval Using the Corners Area Variability Based on Correlogram (코너영역 분산치 기반 코렐로그램을 이용한 형태검출)

  • An, Young-Eun;Lee, Ji-Min;Yang, Won-Ii;Choi, Young-Il;Chang, Min-Hyuk
    • The Journal of the Institute of Internet, Broadcasting and Communication
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    • v.11 no.6
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    • pp.283-288
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    • 2011
  • This paper have proposed an object retrieval using the corners area variability based on correlogram. The proposed algorithm is processed as follows. First, the corner points of the object in an image are extracted and then the feature vectors are obtained. It are rearranged according to the number dimension and consist of sequence vectors. And the similarity based on the maximum of sequence vectors is measured. The proposed technique is invariant to the rotation or the transfer of the objects and more efficient in case that the objects present simple structure. In simulation that use Wang's database, the method presents that the recall property is improved by 0.03% and more than the standard corner patch histogram.

Numerical Simulation of Air Contaminant Dispersion around Urban City (도심지 빌딩주변 자동차 배가스 분산에 관한 연구)

  • Yu, Kyung-Seok;Ryu, Il-Kwang
    • Journal of environmental and Sanitary engineering
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    • v.21 no.2 s.60
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    • pp.11-19
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    • 2006
  • Ran numerical analysis about simplicity warm current model (simple turbulence model) to examine closely distributed laying stress on NOx of contaminant that happen in downtown that building is massed and result is as following. Downtown building is massed and transfer of vehicles can know that frequent area receives greatly effect of building. Pollution displays class kind phenomenon striking in the building, class kind done pollution that show high density being mixed with pollution that happen in vehicles being flowed in again side know can. Divide velocity vector of pollution and density change to ancient city and did numerical analysis. Because NOx is flowed in diversion of water neighborhood by continuous this kind in Godo 20m point, density showed density of $0.0907mg/m^3s$, and can know that Banryuhyeon that Pollution strikes in the building at ancient city 30m point is decreased and approaches with freedom flowing without cooperation (cavity) phenomenon remarkably. Because pollution strikes in the building at distance shaft 383m point, class kind do phenomenon that know appeared notedly Nopeungeonmulgwanat between building pollution density $0.067mg/m^3$ head of a families high appear.

Identification of impact forces on composite structures using an inverse approach

  • Hu, Ning;Matsumoto, Satoshi;Nishi, Ryu;Fukunaga, Hisao
    • Structural Engineering and Mechanics
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    • v.27 no.4
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    • pp.409-424
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    • 2007
  • In this paper, an identification method of impact force is proposed for composite structures. In this method, the relation between force histories and strain responses is first formulated. The transfer matrix, which relates the strain responses of sensors and impact force information, is constructed from the finite element method (FEM). Based on this relation, an optimization model to minimize the difference between the measured strain responses and numerically evaluated strain responses is built up to obtain the impact force history. The identification of force history is performed by a modified least-squares method that imposes the penalty on the first-order derivative of the force history. Moreover, from the relation of strain responses and force history, an error vector indicating the force location is defined and used for the force location identification. The above theory has also been extended into the cases when using acceleration information instead of strain information. The validity of the present method has been verified through two experimental examples. The obtained results demonstrate that the present approach works very well, even when the internal damages in composites happen due to impact events. Moreover, this method can be used for the real-time health monitoring of composite structures.

First-order Generalized Integrator Based Frequency Locked Loop and Synchronization for Three-Phase Grid-connected Converters under Adverse Grid Conditions

  • Luo, Zhaoxu;Su, Mei;Sun, Yao;Liu, Zhangjie;Dong, Mi
    • Journal of Power Electronics
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    • v.16 no.5
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    • pp.1939-1949
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    • 2016
  • This paper presents an alternative frequency adaptive grid synchronization technique named HDN-FLL, which can accurately extract the fundamental positive- and negative-sequence components and interested harmonics in adverse three-phase grid voltage. The HDN-FLL is based on the harmonic decoupling network (HDN) consisting of multiple first order complex vector filters (FOCVF) with a frequency-locked loop (FLL), which makes the system frequency adaptive. The stability of the proposed FLL is strictly verified to be global asymptotically stable. In addition, the linearization and parameters tuning of the FLL is also discussed. The structure of the HDN has been widely used as a prefilter in grid synchronization techniques. However, the stability of the general HDN is seldom discussed. In this paper, the transfer function expression of the general HDN is deduced and its stability is verified by the root locus method. To show the advantages of the HDN-FLL, a simulation comparison with other gird synchronization methods is carried out. Experimental results verify the excellent performance of the proposed synchronization method.

Cloning and Expression of Glucose-1-Phosphate Thymidylyltransferase Gene (schS6) from Streptomyces sp. SCC-2136

  • Han, Ji-Man;Kim, Su-Min;Lee, Hyo-Jung;Yoo, Jin-Cheol
    • Journal of Microbiology and Biotechnology
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    • v.17 no.4
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    • pp.685-690
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    • 2007
  • The deoxysugar biosynthetic gene cluster of Sch 47554/Sch 47555 was cloned from Streptomyces sp. SCC-2136. One of the ORFs, schS6, appeared to encode glucose-1-phosphate thymidylyltransferase, which converts dTTP and glucose-1-phosphate to TDP-D-glucose and pyrophosphate. The dTDP-D-glucose is a key metabolite in prokaryotics as a precursor for a large number of modified deoxysugars, and these deoxysugars are a maj or part of various antibiotics, ranging from glycosides to macrolides. SchS6 was expressed in E. coli vector pSCHS6 and the expressed protein was purified to apparent homogeneity by ammonium sulfate precipitation and Ni-NTA affinity column chromatography. The specific activity of the purified enzyme increased 4.7-fold with 17.5% recovery. It migrated as a single band on SDS-PAGE with an apparent molecular mass of 56kDa. The purified protein showed glucose-1-phosphate thymidylyltransferase activity, catalyzing a reversible bimolecular group transfer reaction. In the forward reaction, the highest activity was obtained with combination of dTTP and ${\alpha}-D-glucose-1-phosphate$, and only 12% of that activity was obtained with the substrates $UTP/{\alpha}-D-glucose-1-phosphate$. In the opposite direction, the purified protein was highly specific for dTDP-D-glucose and pyrophosphate.

Efficiency of transformation mediated by Agrobacterium tumefaciens using vacuum infiltration in rice (Oryza sativa L.)

  • Safitri, Fika Ayu;Ubaidillah, Mohammad;Kim, Kyung-Min
    • Journal of Plant Biotechnology
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    • v.43 no.1
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    • pp.66-75
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    • 2016
  • Agrobacterium-mediated gene transfer has recently been developed to improve rice transformation. In this study, 3 different transformation methods were tested including soaking, co-cultivation, and vacuum infiltration. Agrobacterium tumefaciens GV3101 harboring the binary vector pGreen:: LeGSNOR was used in this experiment. This study aimed to identify the most appropriate method for transferring LeGSNOR into rice. Vacuum infiltration of the embryonic calli for 5 min in Ilpum resulted in high transformation efficiency based on confirmation by PCR, RT-PCR, and qRT-PCR analyses. In conclusion, we described the development of an efficient transformation protocol for the stable integration of foreign genes into rice; furthermore, the study results confirmed that PCR is suitable for efficient detection of the integrated gene. The vacuum infiltration system is a potentially useful tool for future studies focusing on transferring important genes into rice seed calli, and may help reduce time and effort.