• Journal of Microbiology and Biotechnology
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• v.17 no.5
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• pp.728-732
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• 2007

In the present article, a novel fusion expression vector for Escherichia coli was developed based on the pTORG plasmid, a derivative of pET32a. This vector, named pT7MT(GenBank Accession No DQ504436), carries a T7 promoter and it drives the downstream gene encoding Metallothionein 2A(MT2A). There are in-framed multiple cloning sites(MCS) downstream of the MT2A gene. A target gene can be cloned into the MCS and fused to the C-terminal of the MT2A gene in a compatible open reading frame(ORF) to achieve fusion expression. The metal-binding capability of MT2A allows the purification of fusion proteins by metal chelating affinity chromatography, known as $Ni^{2+}$-affinity chromatography. Using this expression vector, we successfully got the stable and high-yield expression of MT2A-GST and MT2A-Troponin I fusion proteins. These two proteins were easily purified from the supernatant of cell lysates by one-step $Ni^{2+}$-affinity chromatography. The final yields of MT2A-GST and MT2A-Troponin I were 30mg/l and 28mg/l in LB culture, respectively. Taken together, our data suggest that pT7MT can be applied as a useful expression vector for stable and high-yield production of fusion proteins.

• Proceedings of the Korean Magnestics Society Conference
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• 1996.10a
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• pp.116-117
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• 1996

• The Pure and Applied Mathematics
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• v.12 no.4 s.30
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• pp.275-287
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• 2005

Geometric invariants are basic tools for geometric processing and computer vision. In this paper, we give a linear approximation for the differentiation of the binormal vector field of a space curve by using the forward and backward differences of discrete binormal vectors. Two kind of discrete torsion, say, back-ward torsion $T_b$ and forward torsion $T_f$ can be defined by the dot product of the (backward and forward) discrete differentiation of binormal vectors that are linear approximations of torsion. Using Frenet formula and Taylor series expansion, we give error estimations for the discrete torsions. We also give numerical tests for a curve. Notably the average of $T_b$ and $T_f$ looks more stable in errors.

• Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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• 2014.05a
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• pp.813-815
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• 2014

Novel baculovirus vector systems including genes of polyhedron promoter, vesicular stomatitis virus G (VSVG), polyA, cytomegalovirus (CMV) promoter, enhanced green fluorescent protein (EGFP), and protein transduction domain (PTD) were constructed. These recombinant baculovirus vector systems were transfected into diverse cells of 293T, HepG2, HFF, and Hur7 cells and compared the effects of gene transfer and expression of these vector systems with control vector. From the result, we confirmed that these recombinant baculovirus vector systems were more excellent than control vector in efficacy of gene transfer and expression.

• 한국생물공학회:학술대회논문집
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• 2000.04a
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• pp.588-591
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• 2000

Serine palmitoyl transferase(SPT) and ceramidase are the key enzymes in sphingolipid biosynthesis. To study sphingolipid metabolism, we have got the 5'-upstream regions of human serine palmitoyl transferase subunit II and acid ceramidase gene by using GenomeWalker kits(Clontech Co.). Human genomic DNA was purified from HT29, human colon canser cell line by using DNAzol. We got several bands after secondary PCR and subcloned them to T7bule vector. Human SPTII promoter which we got was 2690bp but we cut it with Bgl II and vector with Bgl II and BamH I, and subcloned 1782bp to pGL2-enhancer vector and pGL2-basic vector with luciferase reporter gene. Human acid ceramidase promoter which we got were 2028bp and 1034bp and subcloned to pGL2-enhancer vector and pGL2-basic vector. We transfected these promoters to HT29 cell and assayed luciferase activity. For measuring transfection efficiency, pRL-TK vector with seapancy luciferase reproter gene was cotransfected with these promoters.

• Proceedings of the KIPE Conference
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• 2000.07a
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• pp.413-416
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• 2000

본논문에서는 유도전동기 속도 및 토크 제어시 고조파의 왜형률을 감소시키고 디지털 구현이 용이하며 선형제어 영역을 증가시킬 수 있는 스위칭 방식으로 Space voltage Vector PWM(Pulse Width Modulation) 설명하였으며 시뮬레이션하기 위한 user_tool로써 Matlab/Simulink를 이용하여 SVPWM을 구현하는 방법을 설명하였다 결과적으로 개루프 운전 시 유도기에 인가되는 기준전압을 각각 비과변조와 과변조 시 전압비율 변경방식을 사용하여 나타나는 응답으로 가상스위칭 시간 T1, T2, T0 와 속도, 상전류, 토크응답을 알아보았다.

• IMMUNE NETWORK
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• v.3 no.2
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• pp.110-117
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• 2003

Background: The usefulness of DNA vaccine at priming step of heterologous prime-boost vaccination led to DNA vaccine closer to practical reality. DNA vaccine priming followed by recombinant viral vector boosting via systemic route induces optimal systemic immunity but no mucosal immunity. Mucosal vaccination of the reversed protocol (recombinant viral vector priming-DNA vaccine boosting), however, can induce both maximal mucosal and systemic immunity. Here, we tried to address the reason why the mucosal protocol of prime-boost vaccination differs from that of systemic vaccination. Methods: To address the importance of primary immunity induced at priming step, mice were primed with different doses of DNA vaccine or coadministration of DNA vaccine plus mucosal adjuvant, and immunity including serum IgG and mucosal IgA was then determined following boosting with recombinant viral vector. Next, to assess influence of humoral pre-existing immunity on boosting $CD8^+$ T cell-mediated immunity, $CD8^+$ T cell-mediated immunity in B cell-deficient (${\mu}K/O$) mice immunized with prime-boost regimens was evaluated by CTL assay and $IFN-{\gamma}$-producing cells. Results: Immunity primed with recombinant viral vector was effectively boosted with DNA vaccine even 60 days later. In particular, animals primed by increasing doses of DNA vaccine or incorporating an adjuvant at priming step and boosted by recombinant viral vector elicited comparable responses to recombinant viral vector primed-DNA vaccine boosted group. Humoral pre-existing immunity was also unlikely to interfere the boosting effect of $CD8^+$ T cell-mediated immunity by recombinant viral vector. Conclusion: This report provides the important point that optimally primed responses should be considered in mucosal immunization of heterologous prime-boost regimens for inducing the effective boosting at both mucosal and systemic sites.

• Journal of Korea Society of Industrial Information Systems
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• v.12 no.5
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• pp.54-59
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• 2007

We abtain some stability results for a very general differential system using the method of cone valued vector Lyapunov functions and conversely some sufficient conditions for existence of such vector Lyapunov functions.

• Korean Journal of Veterinary Research
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• v.39 no.4
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• pp.786-796
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• 1999

As an attempt to develop an effective control method against theileriosis, recombinant antigen protein was produced. Thirty-two kDa membrane protein(MP) gene of T sergenti was amplified through RT-PCR from extracted total RNA of T sergenti isolated in Chonbuk, Korea. The amplified 869 bp of Korean T sergenti membrane gene was cloned and the base sequences were analyzed. The amplified gene was cloned into E coli expression vector, pQE32 plasmid vector, and the vector was introduced into E coli strain M15 to produce the recombinant membrane protein. For the induction of T sergenti membrane protein(KTs-MP), the plasmid harboring E coli strain M15 were cultured in the presence of IPTG, and the recombinant protein were purified by $Ni^+$-NTA agarose. Then, to confirm the authenticity of the produced membrane protein, molecular weight of expressed recombinant KTs-MP was analyzed by SDS-PAGE and Western blotting. The molecular weight of expressed recombinant protein was 32 kDa as expected. The recombinant KTs-MP was successfully recognized by anti-His Tag antibody, antisera of T sergenti infected cattle and monoclonal antibody of T sergenti membrane protein. Therefore, we concluded that the authentic 32 kDa membrane protein of T sergenti was produced as immunologically recognizable form.

• Journal of the Korean Statistical Society
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• v.24 no.1
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• pp.149-160
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• 1995

We consider the problem of optimal adaptive estiamtion of the euclidean parameter vector $\theta$ of the univariate non-linerar autogressive time series model ${X_t}$ which is defined by the following system of stochastic difference equations ; $X_t = \sum^p_{i=1} \theta_i \cdot T_i(X_{t-1})+e_t, t=1, \cdots, n$, where $\theta$ is the unknown parameter vector which descrives the deterministic dynamics of the stochastic process ${X_t}$ and ${e_t}$ is the sequence of white noises with unknown density $f(\cdot)$. Under some general growth conditions on $T_i(\cdot)$ which guarantee ergodicity of the process, we construct a sequence of adaptive estimatros which is locally asymptotic minimax (LAM) efficient and also attains the least possible covariance matrix among all regular estimators for arbitrary symmetric density.