• 한국미생물·생명공학회지
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• 제36권1호
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• pp.28-33
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• 2008

Streptomyces griseus trypsin (SGT)을 코드하는 sprT 유전자와 그 하류에 존재하는 두 개의 조절 유전자 rsgtR1 및 sgtR2를 동시에 갖고 있는 재조합 벡터 pWHM3-TR1R2를 S. lividans TK24 및 S. griseus IFO 13350에 도입하여, 트립신의 생산성을 더욱 증대시킬 수 있는 배지를 조사하였다. S. lividans TK24/pWHM3-TR1R2의 경우 배양 5일을 기준으로 R2YE에서 가장 높은 생산성(0.74 unit/mL)을 나타냈고, C5/L. (0.66 unit/mL), Livid (0.08 unit/mL), NDSK(0.06 unit/mL) 순으로 나타났다 S. griseus IFO 13350/pWHM3-TR1R2의 경우에는 전반적으로 배양 7일에 트립신 활성이 가장 높았으며, C5/L (1.518 unit/mL), R2YE(1.284 unit/mL), NDSK (0.932 unit/mL), Livid (0.295 unit/mL) 순으로 나타났다. S. griseus IFO 13350/pWHM3-TR1R2를 C5/L 배지에서 7일간 배양한 배양액으로부터 $25%{\sim}60%$ ammonium sulfate 침전, CM-sepharose 및 Sp-sepharose column chromatography를 통하여 트립신을 고순도로 정제할 수 있었다. 최종 purification fold는 6.5배, 순수 정제된 트립신의 specific activity는 69,252 unit/mg, 회수율은 1.4%이었다.

• Bulletin of the Korean Chemical Society
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• 제32권11호
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• pp.3861-3864
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• 2011

Rhodamine B dye (RB) has been introduced into the mesoporous silica (SBA15) and Ag anchored mesoporous silica by applying solution impregnation method. Surface treatment of SBA15 with 3-aminopropyltrimethoxysilane (APTMS) facilitates selective anchoring of the RB molecules on SBA15. The photoluminescence spectra of RB confined within SBA15 indicates higher emission intensity, than that of the RB solid, particularly in the presence of Ag nanoparticles. The significant enhancement in photoluminescence intensity is attributed to the local enhancement of the optical fields near the molecules by interactions with silver plasmons.

• KSBB Journal
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• 제17권1호
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• pp.20-25
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• 2002

We performed a basic experiment for rapid, on-line, real-time measurement of HBsAg by using a surface plasmon resonance biosensor to quantify the recognition and interaction of biomolecules. We immobilized the anti-HBsAg polyclonal antibody to the dextran layer on a CM5 chip surface which was pre-activated by N-hydroxysuccinimide for amine coupling. The binding of the HBsAg to the immobilized antibody was measured by the mass increase detected by the change in the SPR signal. The binding characteristics between HBsAg and its antibody followed typical monolayer adsorption isotherm. When the entire immobilized antibody was interacted, there was no additional, non-specific binding observed, which suggested the biointeraction was very specific as expected and independent of the ligand density. No significant steric hindrance was observed at 17.6 nm/$mm^2$ immobilization density. The relationship between the HBsAg concentration in the sample solution and the antigen bound to the chip surface was linear up to ca. $40\mu\textrm{g}$/mL, which is much wider than that of the ELISA method. It appeared the antigen-antibody binding was increased as the immobilized ligand density increased, but verification is warranted. This study showed the potential of this biosensor-based method as a rapid, simple, multi-sample, on-line assay. Once properly validated, it can serve as a more powerful method for HBsAg quantification replacing the current ELISA method.

• 식물병연구
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• 제22권4호
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• pp.269-275
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• 2016

E. pyrifoliae에 의한 과수 가지검은마름병은 국내에서 1995년 최초 발생이래 2016년까지 꾸준히 발생하여 과수농가에 피해를 주고 있는 세균병해이다. 가지검은마름병 발생 농가의 폐원조치 및 공적방제로 인한 경제적 피해 감소를 위하여, 병징이 관찰되는 이병조직 및 건전조직 내의 병원 세균을 검출하여 효율적 관리방안을 마련하고자 본 연구를 수행하였다. 가지검은마름병원세균의 검출은 genomic DNA 추출과정을 생략한 순수 균총만을 이용하는 colony-PCR을 이용하였으며, 이를 위해 ERIC 지역에서 제작된 가지검은마름병원세균 특이 프라이머 EpSPF/EpSPR 프라이머쌍을 선발하였다. 특이 프라이머를 활용한 colony-PCR 방법으로 2014-2015년 4-10월까지 사과나무 생육기간 동안 가지검은마름병 발생상황을 모니터링한 결과, $25^{\circ}C$ 일 평균 온도 기간인 5월 중순부터 7월 초순까지 발병이 가장 빈번하였다. 발병가지 내 병원세균의 존재유무 검정 결과 병징 부위와 이로부터 20 cm 내 건전조직에서만 병원세균이 지속적으로 검출되었다. 따라서 이미 발생한 가지검은마름병을 효율적으로 관리하기 위해 이병조직과 건전조직 경계 부위로부터 20 cm 이상에서 가지치기를 하는 것이 매우 적절할 것으로 판단된다.

• 대한화장품학회지
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• 제49권1호
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• pp.75-85
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• 2023

본 연구에서는 7 개의 아미노산으로 이루어진 헵타펩타이드의 Lgr5 binding에 따른 인체 모낭 구성 세포의 활성에 대한 영향을 확인하였다. 표면 플라즈몬 공명(surface plasmon resonance, SPR) 시스템을 이용하여 헵타펩타이드가 Lgr5에 결합하는 것을 확인하였다. 인체 모유두세포(human hair follicle dermal papilla cell, HHFDPC)에 헵타펩타이드를 처리한 결과, 농도 의존적인 세포 증식이 나타났으며 β-catenin의 세포 내핵 이동 및 하위 유전자인 LEF1, Cyclin-D1, c-Myc의 발현 증가가 관찰되었다. 그리고 세포 증식 기전 관련 인자인 Akt와 ERK의 인산화 수준이 증가되었으며, 성장인자인 hepatocyte growth factor (HGF), keratinocyte growth factor (KGF), vascular endothelial growth factor (VEGF) 발현이 유도되었다. 또한 인체 모모세포(human hair germinal matrix cell, HHGMC)의 분화 관련 전사 인자와 인체 외모근초세포(human hair outer root sheath cell, HHORSC)의 분화 표지 인자들도 헵타펩타이드 처리 시 높은 발현율을 보였다. 추가적으로 우리는 헵타펩타이드의 인체 모낭줄기세포(human hair follicle stem cell, HHFSC) 분화에 대한 영향을 조사하였다. 그 결과, HHFSC 표지인자들의 mRNA와 단백질 수준이 감소하였고 반면에 분화 표지인자들은 증가하였다. 상기의 결과들은 헵타펩타이드가 인체 모낭 구성 세포에서 Wnt/β-catenin 경로를 촉진시켜 증식 또는 분화를 유도할 수 있음을 보여준다. 이를 토대로 종합해 볼 때, 본 연구의 헵타펩타이드는 모발 성장을 유도하고 탈모 개선에 도움을 줄 수 있는 기능성 원료로 사용될 수 있을 것으로 보인다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제9권2호
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• pp.137-142
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• 2004

In this study, a simple procedure is described for patterning biotin on a glass substrate and then selectively immobilizing proteins of interest onto the biotin-patterned surface. Microcontact printing (CP) was used to generate the micropattern of biotin and to demonstrate the selective immobilization of proteins by using enhanced green fluorescent protein (EGFP) as a model protein, of which the C-terminus was fused to a core streptavidin (cSA) gene of Streptomyces avidinii. Confocal fluorescence microscopy was used to visualize the pattern of the immobilized protein (EGFP-cSA), and surface plasmon resonance was used to characterize biological activity of the immobilized EGFP-cSA. The results suggest that this strategy, which consists of a combination of $\mu$CP and cSA-fused proteins. is an effective way for fabricating biologically active substrates that are suitable for a wide variety of applications. one such being the use in protein-protein assays.

• 한국도서관정보학회지
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• 제31권2호
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• pp.63-91
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• 2000

본 연구에서는 지식경영에 기반한 기업체에서 지식관리를 담당하는 전문도서관의 분석정보기능의 강화 필요성을 제시하였다. 기업체 정보관리 업무단계에 있어서 이용자 요구에 수동적으로 서비스하는 단계에서 수집된 원자료의 내용을 가공 분석하여 이용자 그릅별 목적에 맞는 맞춤정보(customizing information)를 사내 인트라넷을 통해 다양한 형태로 서비스할 수 있는 전문가적 수준의 '분석정보'의 제공에 대해서 연구하였다. 이는 대규모 도서관에서 이루어지는 정보서비스와는 달리 기업체의 이익창출에 기여할 수 있는 조직내 상부 의사결정권자를 핵심 이용자로 설정하여 기업목표를 달성할 수 있는 기업내 지식관리자자로서의 새로운 입지를 포지셔닝 할 수 있을 것이다. 또한 지식관리시스템의 선도적 역할을 수행할 수 있는 중요한 계기가 될 수 있는 구체적인 방안에 대해서 제시하였다.

• 한국진공학회:학술대회논문집
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• 한국진공학회 2013년도 제44회 동계 정기학술대회 초록집
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• pp.110-111
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• 2013

FcBP consisting of 13 amino acids specifically binds to Immunoglobulin G Fc domain. Initially, we utilized this peptide for preparation of antibody chip as a PEG composite for enhanced solubility. After then, the peptide conjugate was immobilized on agarose resin, resulting in highly efficient affinity column for antibody purification. The efficiency was comparable to commercial Protein A column. Recently, this peptide was conjugated with cell penetrating peptide (CPP) on a backbone of GFP, affording antibody transducer, which carries antibody into live cells by simple mixing of antibody and the transducer in cell culture media. Antibody transduction into cells was monitored by live cell imaging. More recently, the FcBP was fused to ferritin cage, which consists of 24 ferritin protein molecules. The FcBP-ferritin cage showed greatly increased binding affinity to human IgG. Its binding was analyzed by QCM and SPR analysis. Finally, it was selectively delivered by Herceptin to SKBR3, a breast cancer cell, over MCF10A, non-tumorigenic cells (Fig. 1). Fig. 1. Fluorescent microscopic images of SKBR3 breast cancer cells (A~C) and MCF10A breast cells (D~F) treated with Cy3-trastuzumab/fFcBP-Pf_Fn complexes. Trastuzumab and FcBP-Pf_Fn, which were labeled with Cy3 (Cy3-trastuzumab) and fluorescein (fFcBP-Pf_Fn), respectively, selectively targeted SKBR3 over MCF10A.

• Journal of Microbiology and Biotechnology
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• 제19권10호
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• pp.1191-1196
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• 2009

The production of Streptomyces griseus trypsin (SGT) by S. griseus IFO13350 transformed with the expression vector pWHM3-TR1R2, containing sprT encoding SGT and the two positive regulatory genes sgtR1 and sgtR2, was investigated in various media. Cultivation in Ferm-0 gave 1.4 times more trypsin activity than in C5/L medium. In addition, replacement of 2% glucose and 1% skim milk in Ferm-0 with 2% dextrin and 1% tryptone (designated Ferm-II) enhanced trypsin activity 4.1-fold. To simplify the purification process, the supernatant from the S. griseus transformant cultured in Ferm-II medium was fractionated with ammonium sulfate (25-55%), then subjected to Hitrap Benzamidine FF affinity column chromatography. The specific activity of SGT purified by one-step chromatography was 69,550 unit/mg protein and the overall purification yield was above 8%, indicating that this method is more effective than those previously reported. Purified SGT was most active at pH 8.0 and $50^{\circ}C$, and it maintained activity between pH 7.0 and 9.0 and at temperatures up to $70^{\circ}C$. These enzymatic properties are very similar to those of authentic eukaryotic trypsin purified from bovine pancreas.

• Journal of Microbiology
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• 제43권spc1호
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• pp.110-117
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• 2005

Salmonella enterica serovar Typhimurium causes human gastroenteritis and a systemic typhoid-like infection in mice. A critical virulence determinant of Salmonella is the ability to invade mammalian cells. The expression of genes required for invasion is tightly regulated by environmental conditions and a variety of regulatory genes. The hilA regulator encodes an OmpR/ToxR family transcriptional regulator that activates the expression of invasion genes in response to both environmental and genetic regulatory factors. Work from several laboratories has highlighted that regulation of hilA expression is a key point for controlling expression of the invasive phenotype. A number of positive regulators of hilA expression have been identified including csrAB, sirA/barA, pstS, hilC/sirC/sprA, fis, and hilD. HilD, an AraC/XylS type transcriptional regulator, is of particular importance as a mutation in hilD results in a 14-fold decrease in chromosomal hilA::Tn5lacZY-080 expression and a 53-fold decrease in invasion of HEp-2 cells. It is believed that HilD directly regulates hilA expression as it has been shown to bind to hilA promoter sequences. In addition, our research group, and others, have identified genes (hilE, hha, pag, and lon) that negatively affect hilA transcription. HilE appears to be an important Salmonella-specific regulator that plays a critical role in inactivating hilA expression. Recent work in our lab has been directed at understanding how environmental signals that affect hilA expression may be processed through a hilE pathway to modulate expression of hilA and the invasive phenotype. The current understanding of this complex regulatory system is reviewed.