• Biotechnology and Bioprocess Engineering:BBE
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• 제3권2호
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• pp.82-86
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• 1998

The maltose binding protein (MBP) fusion protein system is a versatile tool to express and isolate recombinant proteins in E. coli. In this system, MBP fusion proteins are efficiently isolated from whole cell lysate using amylose conjugated agarose beads and then eluted by competition with free maltose. Since MBP is a rather large molecule (∼42 kDa), for further experiments, the MBP part is usually proteolytically cleaved from the fusion protein and subsequently removed by ion-exchange chromatography or rebinding to amylose columns after washing out excess and MBP-bound maltose. In the present study, we have developed an improved method for the removal of cleaved MBP, which is advantageous over conventional methods. In this method, factor Xa cleaved MBP fusion proteins were incubated with Sepharose beads conjugated with MBP specific monoclonal antibodies and then precipitated buy centrifugation, resulting in highly purified proteins in the supernatant.

• Parasites, Hosts and Diseases
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• 제54권4호
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• pp.461-469
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• 2016

Giardia lamblia is a protozoan that causes diarrheal diseases in humans. Cytoskeletal structures of Giardia trophozoites must be finely reorganized during cell division. To identify Giardia proteins which interact with microtubules (MTs), Giardia lysates were incubated with in vitro-polymerized MTs and then precipitated by ultracentifugation. A hypothetical protein (GL50803_8405) was identified in the precipitated fraction with polymerized MTs and was named GlMBP1 (G. lamblia microtubule-binding protein 1). Interaction of GlMBP1 with MTs was confirmed by MT binding assays using recombinant GlMBP1 (rGlMBP1). In vivo expression of GlMBP1 was shown by a real-time PCR and western blot analysis using anti-rGlMBP1 antibodies. Transgenic G. lamblia trophozoites were constructed by integrating a chimeric gene encoding hemagglutinin (HA)-tagged GlMBP1 into a Giardia chromosome. Immunofluorescence assays of this transgenic G. lamblia, using anti-HA antibodies, revealed that GlMBP1 mainly localized at the basal bodies, axonemes, and median bodies of G. lamblia trophozoites. This result indicates that GlMBP1 is a component of the G. lamblia cytoskeleton.

• Toxicological Research
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• 제16권3호
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• pp.221-227
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• 2000

A 6-sulfooxymethylbenzo[a]pyrene (SMBP), the ultimate metabolite of methyl-substituted benzo[a]pyrene (BP), has been found to be carcinogenic in mice. These properties may be attributable to its strong reactivity with cellular macromolecules such as DNA. However, serum and its major constituent albumin attenuated significantly the cytotoxicity and mutagenicity of 5MBP in bacterial and mammalian cell systems. This inhibitory activity of serum against 5MBP-induced cytotoxicity and mutagenicity in Chinese hamster V79 cells appears to be caused by the reduced macromolecular adducts such as DNA and proteins, but serum failed to reduce 5MBP binding to naked calf thymus DNA. A number of proteins in the serum could act as nucleophiles that are able to intercept reactive chemicals through covalent binding. Albumin present in the plasma seems to be one of major components responsible for direct binding with 5MBp, thereby reducing its reactivity to genetic materials. We here determined which fraction is preferential for 5MBP binding through fractionation of 5MBP-treated serum with ammonium sulfate. The albumin-containing fraction had slightly more affinity for 5MBP than the immunoglobulin-containing fraction. Our results indicate that the covalent modification of plasma proteins may reduce 5MBP-induced damage.

• The Korean Journal of Ecology
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• 제21권3호
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• pp.225-231
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• 1998

The studies on the potentiality of biomonitoring heavy metal pollution in coastal region of industrial complex were performed to investigate the heavy metal accumulation and induction of metal-binding protein (MBP) as detoxification process using Rumex maritimus. Bioconcentration in organs and MBP in root of R. maritimus was investigated for the research of the tolerance of heavy metals. The bioconcentration of cadmium and zinc in organs showed 3.6-8.0 times in root higher than in shoot, so it was found that heavy metal accumulated selectively in root. MBP increased absorbance in 254 nm and decreased in 280 nm, because it was composed of high cystein content and low aromatic acids, so absorbance had large difference between 254 nm and 280 nm. The existence of MBP in the 10-20 fraction was ascertained with anion exchange chromatography and it was identified that concentration of heavy metal increased according as an exposure concentration of medium increased in QAE Sephadex A-25 elution profile. These results suggested that MBP could play a role in biomarker determining the bioconcentration of plant. This study demonstrated a possibility that removal ability of heavy metal of R. maritimus resulted from detoxification process and MBP could be utilized as a biomarker of heavy metal pollution.

• 산업경영시스템학회지
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• 제10권16호
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• pp.53-61
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• 1987

This study is on the Management by Policy-Deployment(MBP) a Japanese-devised management system which recently call a widespread awareness and applications among major Korean Industries. The principal nature of MBP was compared with Management by Objectives(MBO) as these two systems share similarities in basic nature. Then the major application problems of MBP encountered by the practicing managers were encountered and examined In relation to the Japanese firms organizational culture.

• 생명과학회지
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• 제25권4호
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• pp.385-389
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• 2015

선행연구에서 카스파제-3 효소가 DEVD 기질을 완전히 절단하는 반면 IETD 기질은 DEVD의 약 50%정도만을 부분적으로 분해한다는 사실을 보고하였다. 본 연구에서는 정제된 폴리-단백질이 카스파제-3 단백질 분해효소의 기질에 따른 차별적인 분해활성에 의해 프로세싱 되는 양상을 분석하였다. 모델 단백질로서 GST, MBP, RFP 세 종류의 단백질을 DEVD 및 IETD 펩타이드로 연결시킨 폴리-단백질을 제작하였으며, 폴리-단백질은C-말단에 6개의 히스티딘 태그가 결합되도록 클로닝 되었다. IMAC 크로마토그래피를 이용하여 분리 및 정제된 재조합 단백질은 SDS-PAGE 분석을 통해 분자량이 약 93 kDa으로 나타났으며, 카스파제-3 효소의 처리에 의해 각각 MBP:RFP, MBP 그리고 GST 3종류의 단백질 절편으로 절단 및 분리되었다. 이 연구를 통해 단백질 분해효소와 기질 간의 반응성의 차이를 이용하여 폴리-단백질을 정량적으로 프로세싱 할 수 있다는 예를 보여주었다.

• 분석과학
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• 제15권4호
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• pp.329-334
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• 2002

TBP 내의 분해산물인 $DBP^-$ 및 $MBP^{2-}$의 정량을 위하여 이온크로마토그래피를 이용하였으며, AS4A-SC(Dionex) 분리관과 $Na_2CO_3$/NaOH 용리액을 사용하여 $DBP^-$, $MBP^{2-}$, $F^-$, $Cl^-$, ${NO_2}^-$, ${NO_3}^-$, ${SO_4}^{2-}$ 및 ${PO_4}^{3-}$의 머무름거동을 살펴보았다. 2 mM $Na_2CO_3$/1 mM NaOH 용리액에서 이들의 분리가 가장 효율적이었으며, 15분 이내에 분리되었다. $5{\mu}g/mL$ - $100{\mu}g/mL$의 농도범위가 측정에 적합하였으며, 이 범위에서 검량곡선은 $r^2$=0.999로 좋은 상관관계를 보여주었다. 시료량 $25{\mu}L$를 취할경우 $DBP^-$의 검출한계는 $1{\mu}g/mL$이고, $MBP^{2-}$의 검출한계는 $0.5{\mu}g/mL$ 이었다.

• Journal of Microbiology and Biotechnology
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• 제8권1호
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• pp.34-41
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• 1998

A method was developed for the controlled expression of an antimicrobial peptide magainin in Escherichia coli. A series of concatemeric magainin genes was constructed with a gene amplification vector, and fused to the 3'end of malE gene encoding the affinity ligand, E. coli maltose-binding protein (MBP). The construct directed the synthesis of the fusion protein with the magainin polypeptide fused to the C-terminus of MBP. The fusion protein was expressed in a tightly regulatable expression system which was under the control of an invertible promoter. The MBP-fused magainin monomer was expressed efficiently. However, the expression level of the MBP-fused magainin in E. coli decreased with the increasing size of multimers possibly because of the transcription and translation inhibition by the multimeric peptides. After purification using an amylose affinity column, the fusion protein was digested by factor Xa at a specific cleavage site between the monomers. The recombinant magainin had an antimicrobial activity identical to that of synthetic magainin. This experiment shows that a biologically active, antimicrobial peptide magainin can be produced by fusing to MBP, along with a promoter inversion vector system.

• 한국정보통신학회논문지
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• 제9권1호
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• pp.45-57
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• 2005

본 논문에서는 종단간 네트워크 시스템에서 전송 율 기반하의 혼잡 제어를 제안하여 양방향 TCP 연결의 성능을 개선한다. TCP의 패킷과 승인들은 종단 시스템의 공통 버퍼를 공유하기 때문에 소스노드에 패킷과 승인이 집단적으로 도착되는 승인 압축 결과를 초래함으로서 불공성과 처리 율이 감소한다. 양방향 트래픽에 의한 처리율 감소는 매우 중요하다. TCP 비율제어를 이용한 백그라운드 트래픽이 2.5Mbps에서 5.0Mbps로 대역폭이 높아짐으로서 중간 노드의 혼잡이 어느 정도 해소되어 2.5Mbps에서 나타낸 것보다 왕복 지연 시간에 대한 처리 율이 적어짐으로서 연결의 효율성이 증가한다. 또한 백그라운드 트래픽을 7.5Mbps로 설정한 지터 처리 율에 대한 시뮬레이션 결과는 혼잡을 제어함으로서 2.5Mbps나 5.0Mbps에 발생한 지터 처리 율보다 개선됨으로서 제안된 비율 제어 방식의 연결 효율성의 성능 개선이 이루어짐을 알 수 있다.

• 한국정보통신학회논문지
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• 제15권3호
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• pp.639-644
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• 2011

초고속 정보 통신망의 발달로, 현재 데이터 통신시스템은 가입자 상호간에 데이터를 빠르고, 정확하게 교환 할 수 있도록 하였다. 본 논문에서는 통신 시스템에 사용되는 여러 가지 Logic중 가장 기초가 되는 LVTTL(Low Voltage Transistor Transistor Logic)을 이용하여 데이터 전송특성 분석을 위한 시스템을 설계하고 데이터 전송속도의 변화따른 LVTTL의 특성을 측정 분석하였다. 현재 시스템에 필요한 전송 라인의 길이가 30cm이기 때문에, 우리는 현재 시스템에 필요한 전송 라인의 길이에 따라 LVTTL 데이터 전송 특성을 분석했다. LVTTL의 신호 레벨은 10Mbps일 경우 3V, 50Mbps일 경우 2.2V, 100Mbps일 경우 2V, 125Mbps일 경우 1.5V, 150Mbps일 경우 1.4V이다. 전송선로의 길이가 30cm, 데이터 전송속도 100Mbps까지 안정하게 보냄을 알 수 있었다.