• Journal of Microbiology and Biotechnology
• /
• 제12권1호
• /
• pp.65-70
• /
• 2002

A TGF-${\beta}1$/GFP monomeric fusion protein was cloned from pPK9A and pGFP-Cl plasmid by PCR amplification. The fusion protein was expressed in a $Bac-To-Bac^{TM}$ baculovirus expression system. A 45 kDa fusion protein was purified using an Ni-NTA column with 300 mM imidazol from a cell lysate infected with recombinant viruses for 72 h post-infection. The fusion protein cross-reacted with the commercial $TGF-{\beta}1$ polyclonal Ab as well as Ab raised against a precursor, monomeric $TGF-{\beta}1$, and GFP. The binding activity of the fusion protein with a $TGF-{\beta}1$ receptor was examined. Fluorescence was observed in Mv1Lu cells, yet not in insect cells treated with the fusion protein. No fluorescence was detected in Mv1Lu cells incubated with the fusion protein treated with Ab prior to the binding reaction, or with GFP alone, thereby indicating that the binding of the fusion protein was specific to $TGF-{\beta}1$ with a receptor.

• Journal of Microbiology and Biotechnology
• /
• 제8권6호
• /
• pp.669-675
• /
• 1998

Taq DNA polymerase exhibits a sizable drawback compared to the other thermophilic DNA polymerases in that it demonstrates lower proof-reading activity due to the deficiency of 3'-5'exonuclease activity. A study was undertaken to improve the 3'-5' exonuclease activity in the PCR of Taq DNA polymerase. The three-dimensional structural alignment of the polymerase and 3'-5' exonuclease domains from the pol I family DNA polymerases explains why Taq DNA polymerase has just a background level of 3'-5'exonuclease activity. A comparison indicated that the two polymerase domains are very similar in primary and tertiary conformations, even though Taq DNA polymerase carries a much shorter 3'-5'exonuclease domain than that of E. coli DNA polymerase I. Those two polymerase domains were interchanged between Taq DNA polymerase and E. coli DNA polymerase I. The 3'-5' exonuclease domain from E. coli DNA polymerase I was separated and pasted into the polymerase domain of Taq DNA polymerase I, which resulted in a functional fusion-Stoffel fragment. The 3'-5'exonuclease activity of the fusion-Stoffel fragment increased up to 48% of the value of the Klenow fragment, while that of Taq DNA polymerase remained at 6.0% of the Klenow fragment.

• 생약학회지
• /
• 제34권1호통권132호
• /
• pp.28-32
• /
• 2003

In order to search for the anti-HIV agents from natural products, eighty MeOH extracts of medicinal plants were applied to a syncytia formation inhibition assay which is based on the interaction between the HIV-1 envelope glycoprotein gp120/gp41 and the cellular membrane protein CD4 of T lymphocytes. Among them, Ailanthus altissima showed a potent virus-cell fusion inhibitory activity. Repeated column chromatoghaphy of the methylene chloride fraction of A. altissima afforded compounds 1$({\beta}-sitosterol-3-O-{\beta}-D-glucoside)$, 2(tetramethoxycoumarin), and 3(ocotillone). Virus-cell fusion inhibitory activity of compound 3(ocotillone) was $70.76{\pm}4.09%$ at the concentration of $100\;{\mu}g/ml$.

• Asian-Australasian Journal of Animal Sciences
• /
• 제30권4호
• /
• pp.576-584
• /
• 2017

Objective: To generate recombinant Bacillus subtilis (B. subtilis) engineered for expression of porcine ${\beta}-defensin-2$ (pBD-2) and cecropin P1 (CP1) fusion antimicrobial peptide and investigate their anti-bacterial activity in vitro and their growth-promoting and disease resisting activity in vivo. Methods: The pBD-2 and CP1 fused gene was synthesized using the main codons of B. subtilis and inserted into plasmid pMK4 vector to construct their expression vector. The fusion peptide-expressing B. subtilis was constructed by transformation with the vector. The expressed fusion peptide was detected with Western blot. The antimicrobial activity of the expressed fusion peptide and the recovered pBD-2 and CP1 by enterokinase digestion in vitro was analyzed by the bacterial growth-inhibitory activity assay. To analyze the engineered B. subtilis on growth promotion and disease resistance, the weaned piglets were fed with basic diet supplemented with the recombinant B. subtilis. Then the piglets were challenged by enteropathogenic Escherichia coli (E. coli). The weight gain and diarrhea incidence of piglets were measured after challenge. Results: The recombinant B. subtilis engineered for expression of pBD-2/CP1 fusion peptide was successfully constructed using the main codons of the B. subtilis. Both expressed pBD-2/CP1 fusion peptide and their individual peptides recovered from parental fusion peptide by enterokinase digestion possessed the antimicrobial activities to a variety of the bacteria, including gram-negative bacteria (E. coli, Salmonella typhimurium, and Haemophilus parasuis) and grampositive bacteria (Staphylococcus aureus). Supplementing the engineered B. subtilis to the pig feed could significantly promote the piglet growth and reduced diarrhea incidence of the piglets. Conclusion: The generated B. subtilis strain can efficiently express pBD-2/CP1 fusion antimicrobial peptide, the recovered pBD-2 and CP1 peptides possess potent antimicrobial activities to a variety of bacterial species in vitro. Supplementation of the engineered B. subtilis in pig feed obviously promote piglet growth and resistance to the colibacillosis.

• 한국컴퓨터정보학회논문지
• /
• 제24권5호
• /
• pp.59-66
• /
• 2019

According to the KT telecommunication statistics, people stayed inside their houses on an average of 11.9 hours a day. As well as, according to NSC statistics in the united states, people regardless of age are injured for a variety of reasons in their houses. For purposes of this research, we have investigated an abnormal event detection algorithm to classify infrequently occurring behaviors as accidents, health emergencies, etc. in their daily lives. We propose a fusion method that combines three classification algorithms with vision pattern, audio pattern, and activity pattern to detect unusual user events. The vision pattern algorithm identifies people and objects based on video data collected through home CCTV. The audio and activity pattern algorithms classify user audio and activity behaviors using the data collected from built-in sensors on their smartphones in their houses. We evaluated the proposed individual pattern algorithm and fusion method based on multiple scenarios.

• 대한지역사회영양학회지
• /
• 제17권4호
• /
• pp.376-389
• /
• 2012

The purpose of this study was to develop a fusion-food cooking program and apply that to sixth grade elementary school students, and to investigate its' effects on their knowledge, preferences, and perceptions of Korean traditional foods. The program focused on ten components. Students learned the general features of Korean traditional foods and Western foods in the 1st lesson. They learnt about kinds, nutrition value, and histories of kimchi, Tteok (rice cake), and Jeon (pan fried food), and cooked two fusion-foods of kimchi, Tteok, and Jeon each. As a result of learning about those food items, the students advanced their knowledge (p < 0.001) of kimchi, Tteok, and Jeon. Their preferences for Tteok (p < 0.05) and Jeon (p < 0.01) were increased, but those for kimchi remained unchanged. The interest (p < 0.05) and preferences (p < 0.01) for the general Korean traditional foods were increased. In conclusion, our results suggested that the fusion-food cooking program had the ability to improve elementary school students' perceptions of Korean traditional foods by increasing their knowledge, preferences, and interest in them. Furthermore it was considered that the program could help students understand the value of Korean traditional foods and in turn may encourage them to consume such food items more frequently. Since the fusion-food cooking activity program can be a good learning program as shown by the results of this study, more fusion-foods cooking activity programs, which are not discussed in this study, should be evaluated and developed in the future.

• 한국환경과학회:학술대회논문집
• /
• 한국환경과학회 2020년도 정기학술대회 발표논문집
• /
• pp.215-215
• /
• 2020

• Bulletin of the Korean Chemical Society
• /
• 제27권2호
• /
• pp.325-328
• /
• 2006

• 한국미생물학회:학술대회논문집
• /
• 한국미생물학회 2001년도 추계학술대회
• /
• pp.97-101
• /
• 2001

The fusion between viral envelope and target cell membrane is a central step of viral infection, and the fusion proteins located at viral envelope mediate such process. Gp41 of HIV is one of the fusion proteins whose structure and mechanism of membrane fusion had been extensively studied. Functionally important motives of gp41 are the N-terminus fusion peptide, the coiled-coil and the membrane proximal C-peptide regions. The role of these regions during the fusion process had been thoroughly examined. Specially, insertion of the fusion peptide into membrane and conformational change of the coiled-coil and C-peptide regions are assumed to be critical for the fusion mechanism. In addition, the coiled-coil region has been shown to interact with membrane, and the C-peptide region regulates the interaction in a dose dependent manner. Furthermore, fusion defective mutations of the coiled-coil region dramatically changed its binding affinity to membrane. These results suggested that the membrane binding property of the coiled-coil region is important for the fusion activity of gp41, and such property could be modulated by the interaction with the C-peptide region.

• 한국식품과학회지
• /
• 제25권4호
• /
• pp.366-372
• /
• 1993

본 연구는 미림 제조시 미림의 품질을 높이기 위한 방법의 하나로 세포 원형질체의 융합체 의해 ACPase 활성이 높은 균주를 얻으려고 시도하였다. 돌연변이 방법으로 acid carboxypeptidase(ACPase) 활성이 높은 A. oryzae 9-12와 A. shirousamii IFO 6082-60을 선별하여 사용하였다. A. oryzae 9-12와 A. shirousamii IFO 608260의 원형질체의 형성조건은 chitinase (10mg/ml), cellulase(10mg/ml) 및 zymolase(20T(5mg/ml)등의 효소 혼합용액에서 $30^{\circ}C$에서 4시간 반응시켰을 때 가장 높았으며, pH6.0일 때, 안정제로서는 0.6M KCl 및 무기 염류로서는 0.01M $CaCl_2\;2H_2O$가 가장 높았고, 30% PEG 6,000이 가장 좋았다. 이때의 융합빈도는 0.71%였다. 융합주의 ACPase의 활성은 F-50이 20,800unit/g으로 변이주보다 1.4배 높았다.