• 한국정보통신학회:학술대회논문집
• /
• 한국정보통신학회 2017년도 춘계학술대회
• /
• pp.700-703
• /
• 2017

베큘로바이러스 벡터가 uroplakin II promoter, polyhedron promoter, vesicular stomatitis virus G (VSVG), enhanced green fluorescent protein (EGFP), protein transduction domain (PTD) 등의 유전자로 재조합 되었다. 이렇게 재조합된 베큘로바이러스 벡터들은 여러가지 세포주에 감염을 시켰다. 우리는 이 재조합 벡터와 다른 대조 벡터를 비교하여 유전자 전달과 유전자 발현을 비교하였다. 그 결과 이 재조합 베큘로바이러스 벡터는 대조 벡터에 비하여 유전자 전달과 유전자 발현의 효율이 우수한 것으로 나타났다.

• 한국정보통신학회:학술대회논문집
• /
• 한국정보통신학회 2016년도 춘계학술대회
• /
• pp.711-714
• /
• 2016

재조합 베큘로바이러스는 cytomegalovirus (CMV) promoter, polyhedron promoter, vesicular stomatitis virus G (VSVG), enhanced green fluorescent protein (EGFP), protein transduction domain (PTD) 유전자로 구성된다. 본 재조합 베큘로바이러스 벡터는 세포주와 조직에 감염시켰고 그 결과 다른 벡터 시스템에 비교하여 재조합된 유전자의 전이와 유전자 발현에 있어서 새로운 가능성이 발견되었다. 본 재조합 베큘로바이러스 시스템의 유전자의 전이와 발현의 효율은 타 벡터시스템 보다 우수하였다.

• International Journal of Industrial Entomology and Biomaterials
• /
• 제8권2호
• /
• pp.145-149
• /
• 2004

The expression of a fusion protein comprised of the B. thuringiensis crystal protein, Cry1Ac, and enhanced green fluorescent protein (EGFP) in Escherichia coli XLl-blue was examined. Three recombinant plasmids were transformed into E. coli XL1-blue and named as ProAc/Ec, MuEGFP/Ec and ProMu-EGFP/Ec, respectively. All transformants were observed by light and fluorescence microscopy at mid-log phase. The expression in E. coli transformants, ProMu-EGFP/Ec and MuEGFP/Ec, exhibited bright enough fluorescence to be observed. Furthermore, ProMu-EGFP/Ec produced fluorescent inclusions, which may have been recombinant crystals between EGFP and Cry1Ac while MuEGFP/Ec expressed soluble EGFP in cell. In SDS-PAGE, ProAc/Ec had 130 kDa crystal protein band and MuEGFP/Ec had thick 27 kDa EGFP band. However, ProMu-EGFP/Ec had about 150 kDa fusion protein band. Accordingly, these results indicated that a fusion protein between the B. thuringiensis crystal protein and a foreign protein under the lacZ promoter was successfully expressed as granular structure in E. coli. It is suggested that the E. coli expression system by N-terminal fusion of B. thuringiensis crystal protein may be useful as excellent means for fusion expression and characterization of B. thuringiensis fusion crystal protein.

• 한국정보통신학회논문지
• /
• 제18권8호
• /
• pp.2017-2022
• /
• 2014

폴리히드론 프로모터, 수포성구내염 바이러스G, 폴리A, 사이토메가바이러스 프로모터, 강화녹색형광단백질, 단백질전달부위 유전자 등을 포함한 새로운 베큘로바이러스 시스템이 제조되었다. 본 재조합벡터 시스템은 인간 섬유아세포에 적용하여 시험하였고 재조합된 유전자의 전달과 유전자 발현을 대조 벡터시스템과 비교하였다. 본 연구로부터 새롭게 제작된 본 베큘로바이러스 시스템이 유전자의 전이와 유전자 발현 면에서 대조 벡터시스템 보다 고효율을 나타내었다.

• 대한의생명과학회지
• /
• 제7권4호
• /
• pp.167-172
• /
• 2001

Nebulin is an approximately 700 kDa filamentous protein in vertebrate skeletal muscle. It binds to the Z line and also binds side-by-side to the entire thin actin filament in a sarcomere. The correlation of nebulin size with thin filament length have led to the suggestion that nebulin acts as a molecular ruler for the length of thin filaments. The C-terminal part of human nebulin is anchored in the sarcomeric Z-disk and contains an SH3 domain. SH3 domains have been identified in an ever-increasing number of proteins important for a wide range of cellular processes, from signal transduction to cytoskeleton assembly and membrane localization. However, the exact physiological role of SH3 domains remains, in many cases, unclear. To explore the role of nebulin SH3 in the cytoskeletal rearrangement that accompanies myoblast differentiation, we transfected sense and antisense nebulin SH3 domain fused to enhanced green fluorescent protein in myoblast. Cells expressing nebulin SH3 fragment showed decrease of cell-cell adhesion, and cells transfected with antisense nebulin SH3 gene showed a rounded cell morphology and loss of cell-matrix adhesion. No alteration in cell shape and differentiation were observed in control cells expressing enhanced green fluorescent protein. Perturbation of nebulin altered the cell shape and disrupted cell adhesion in myoblast, demonstrating that nebulin can affect cytoskeleton rearrangement.

• Journal of Microbiology and Biotechnology
• /
• 제22권9호
• /
• pp.1271-1278
• /
• 2012

Genetic fusion of two proteins frequently induces beneficial effects to the proteins, such as increased solubility, besides the combination of two protein functions. Here, we study the effects of the bacterial surface layer protein SgsE from Geobacillus stearothermophilus NRS 2004/3a on the folding of a C-terminally fused enhanced green fluorescent protein (EGFP) moiety. Although GFPs are generally unable to adopt a functional confirmation in the bacterial periplasm of Escherichia coli cells, we observed periplasmic fluorescence from a chimera of a 150-amino-acid N-terminal truncation of SgsE and EGFP. Based on this finding, unfolding and refolding kinetics of different S-layer-EGFP chimeras, a maltose binding protein-EGFP chimera, and sole EGFP were monitored using green fluorescence as indicator for the folded protein state. Calculated apparent rate constants for unfolding and refolding indicated different folding pathways for EGFP depending on the fusion partner used, and a clearly stabilizing effect was observed for the SgsE_C fusion moiety. Thermal stability, as determined by differential scanning calorimetry, and unfolding equilibria were found to be independent of the fused partner. We conclude that the stabilizing effect SgsE_C exerts on EGFP is due to a reduction of degrees of freedom for folding of EGFP in the fused state.

• 한국광학회지
• /
• 제35권1호
• /
• pp.30-34
• /
• 2024

실크 단백질과 녹색 형광 단백질을 결합한 순수 단백질 기반 녹색 형광체의 구현과, 산화티타늄 나노 입자에 의한 형광 증폭 현상을 보고한다. 실크 단백질은 인간에게 다양한 이점을 제공하는 소재로서, 다양한 마이크로/나노 구조 형성 가능성과 투명한 성질을 가지고 있어 광학소재로서의 활용 가능성 또한 높다. 본 연구에서 제작한 소재는 강한 녹색 형광을 띠고 있으며, 산화티타늄 나노 입자에 의해 녹색 형광이 7.5배 증폭되는 것을 확인하였다.

• Journal of Microbiology and Biotechnology
• /
• 제33권12호
• /
• pp.1681-1691
• /
• 2023

Flavin mononucleotide-binding proteins or domains emit cyan-green fluorescence under aerobic and anaerobic conditions, but relatively low fluorescence and less thermostability limit their application as reporters. In this work, we incorporated the codon-optimized fluorescent protein from Chlamydomonas reinhardtii with two different linkers independently into the redox-responsive split intein construct, overexpressed the precursors in hyperoxic Escherichia coli SHuffle T7 strain, and cyclized the target proteins in vitro in the presence of the reducing agent. Compared with the purified linear protein, the cyclic protein with the short linker displayed enhanced fluorescence. In contrast, cyclized protein with incorporation of the long linker including the myc-tag and human rhinovirus 3C protease cleavable sequence emitted slightly increased fluorescence compared with the protein linearized with the protease cleavage. The cyclic protein with the short linker also exhibited increased thermal stability and exopeptidase resistance. Moreover, induction of the target proteins in an oxygen-deficient culture rendered fluorescent E. coli BL21 (DE3) cells brighter than those overexpressing the linear construct. Thus, the cyclic reporter can hopefully be used in certain thermophilic anaerobes.

• Reproductive and Developmental Biology
• /
• 제32권1호
• /
• pp.15-20
• /
• 2008

Pronuclear DNA microinjection has been the most universal method in transgenic animal production but its success rate of transgenesis in mammals are extremely low. To address this long-standing problem, we used retrovirus- and lentivirus-based vectors carrying the enhanced green fluorescent protein (EGFP) gene under the control of ubiquitously active cytomegalovirus (CMV) promoter to deliver transgenes to bovine embryos. The rate of transgenesis was evaluated by counting EGFP positive blastocysts after injection of concentrated virus stock into the perivitelline space of the bovine oocytes in metaphase II. Among two different types of lentivirus vectors derived from FIV (feline immunodeficiency virus) and HIV (human immunodeficiency virus), the former scored the higher gene transfer efficiency; almost 100% of the blastocysts developed from the oocytes infected with FIV-based vector were EGFP positive. As for the vectors derived Com HIV lentivirus, the transgenesis rate of the blastocysts was reduced to 39%.

• 한국응용곤충학회:학술대회논문집
• /
• 한국응용곤충학회 2002년도 추계 합동 특강 및 학술발표대회
• /
• pp.154-154
• /
• 2002