• Title/Summary/Keyword: foreign protein

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Toll-like receptor and silk sericin for tissue engineering

  • Kim, Seong-Gon;Kweon, HaeYong;Jo, You-Young
    • International Journal of Industrial Entomology and Biomaterials
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    • v.42 no.1
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    • pp.1-6
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    • 2021
  • Toll-like receptor (TLR) is responsible for the recognition of foreign protein. Accordingly, TLR is mainly expressed in the immune associated cells. When foreign protein such as silk sericin is considered for the graft, the response of TLR should be considered. TLR is not all or none responsive receptor. TLR can be activated differently by the intensity of the input. Silk sericin is easily fragmented. The protein conformation of silk sericin is different to the degumming method. TLR response to silk sericin may be different to the degumming method. Consequently, objective tailored extraction method should be investigated and developed.

Osmoprotective Effect of Glycine Betaine on Foreign Protein Production in Hyperosmotic Recombinant Chinese Hamster Ovary Cell Cultures Differs among Cell Lines

  • Ryu, Jun-Su;Kim, Tae-Gyeong;Jeong, Ju-Yeong;Lee, Gyun-Min
    • 한국생물공학회:학술대회논문집
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    • 2000.04a
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    • pp.313-316
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    • 2000
  • When 3 recombinant Chinese hamster ovary (rCHO) cell lines, CHO/dhfr-B-22-4, $CS13-1.00^{\ast}$ and $CSl3-0.02^{\ast}$, were cultivated in hyperosmolar media resulting from NaCl addition, their specific foreign protein productivity increased with medium osmolality. Glycine betaine was found to have a strong osmoprotective effect on all 3 rCHO cell lines. Inclusion of 15 mM glycine betaine in hyperosmolar medim enabled rCHO cell lines to grow at 557-573 mOsm/kg where they could not grow in the absence of glycine betaine. However, effect of glycine betaine inclusion in hyperomolar medium on foreign protein production differed among rCHO cell lines. CHO/dhfr-B22-4 cells retained enhanced specific human thrombopoietin (hTPO) productivity in the presence of glycine betaine, and thereby, the maximum hTPO titer obtained at 573 mOsm/kg was increased by 72% over that obtained in the control culture with physiological osmolality (292 mOsm/kg). On the other hand, enhanced specific antibody productivity of $CSl3-1.00^{\ast}$ and $CSl3-0.02^{\ast}$ at elevated osmolality decreased significantly in the presence of glycine betaine at a cost of the recovery of cell growth. As a result, the maximum antibody titer at 557 mOsm/kg was similar to that obtained in the control culture with physiological osmolality. Taken together, efficacy of the simultanous use of hyperosmotic pressure and glycine betaine as a means to improve foreign protein production was variable among different rCHO cell lines.

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Expression of human lactoferrin N-lobe in Pichia pastoris and its antibacterial activity (Pichia pastoris에서 사람 락토페린 N-lobe의 발현과 항균활성)

  • Won, Su-Jin;Jo, Jae-Hyung;Kim, Seung-Hwan;Kwon, Hyuk-Jin;Lee, Hyune-Hwan
    • Korean Journal of Microbiology
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    • v.51 no.3
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    • pp.271-279
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    • 2015
  • Lactoferrin (LF) is a multifunctional, iron-binding glycoprotein found in physiological secretions of mammals. LF shows antibacterial, antiviral and antifungal activities. In the present study, a gene encoding the N-terminal lobe of human lactoferrin (hLF) was isolated, cloned and expressed in methylotrophic yeast, Pichia pastoris. The recombinant hLF-N (rhLF-N) protein was secreted into the culture medium at the level of $458{\mu}g/ml$ in 3 L fermentor. The size of purified hLF-N was estimated as 35 kDa when analyzed by SDS-PAGE and western blotting. The rhLF-N was further confirmed by immunodiffusion using the anti-hLF polyclonal antibody. The expression profile analysis by qRT-PCR showed that the relative mRNA expression of rhLF-N was maximal after 2-3 days of methanol induction and reduced gradually at 4 days. The purified rhLF-N showed broad antibacterial activities against the pathogens such as Staphylococcus aureus, E. coli, Pseudomonas aeruginosa, Burkholderia cepacia, and Salmonella typhimurium. However, rhLF-N showed relatively lower activity when compared to peptides derived from LF. In spite of this weak activity, the rhLF-N expressed in P. pastoris might be more advantageous for the industrial application, because rhLF-N is secreted into the culture medium and the production can also be increased by optimization of culture conditions.

Home-built Solid-state NMR Probe for Membrane Protein Studies

  • Kim, Yong-Ae;Hwang, Jung-Hyun;Park, Jae-Joon
    • Bulletin of the Korean Chemical Society
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    • v.24 no.9
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    • pp.1281-1283
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    • 2003
  • Proteins in highly oriented lipid bilayer samples are useful to study membrane protein structure determination. Planar lipid bilayers aligned and supported on glass slide were prepared. These stack of glass slide with planar lipid bilayers are not well fit for commercial solid-state NMR probe with round coil. Therefore, homebuilt solid-state NMR probe was built and used for a stack of thin glass plates and RF coil is wrapping directly around the flat square sample. The overall filling factor of the coil is much better and the large surface area enhances the extent to orientation by providing uniform environments for the phospholipids and the high ratio of circumference to area reduces edge effects. $^1H\;and\;^{15}N$ double resonance probe for 400 MHz NMR (9.4T) with a flat coil (coil size: 11 mm ${\times}$ 20 mm ${\times}$ 4 mm) is constructed and tested.

Chemical Quality Evaluation of Commercial Infant Formulas (시판 조제 분유의 화학적 품질 평가)

  • 한정은;홍윤호
    • Food Science of Animal Resources
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    • v.21 no.1
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    • pp.56-63
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    • 2001
  • In this study, three domestic and one foreign formulas for the infants up to 5 month old were examined to detect chemical changes such as pH, reactive sulfhydryl groups(RSH) content, 5-hydroxymethylfurfral(HMF) content, available lysine content, electrophoresis, and surface color caused by heat treatment for long term storage. In the SDS-polyacrylamide gel electrophoresis, A and B products showed similar pattern, while C product had a clearly distinguishable $\beta$-lacto-globulin band, but in casein, only D product showed a few strong casein band. RSH content, which indicate the extent of whey protein denaturation, ranged from 4.40 to 5.93 mmole/g protein. HMF content. which indicate the extent of Maillard reaction, ranged from 192 to 432 $\mu$mole/100g in formulas. B product showed the highest RSH and HMF content. Available lysine content ranged from 31 to 46 mg/g protein. Among them D product contain the highest available lysine content and others showed no significant difference. In conclusion, the domestic infant formulas showed higher RSH and HMF content than the foreign product and the available lysine content of the domestic products were lower than of the foreign product.

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The Production of Foreign Protein in Mouse L Cell (Mouse L Cell에서의 외래 유전자 유래 단백질의 생산)

  • ;Tadamitsu Kishimoto
    • Microbiology and Biotechnology Letters
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    • v.21 no.5
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    • pp.421-427
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    • 1993
  • Some interleukin 6 (IL-60 transcription control factors were resported as the regulator of IL-6 expression. A nuclear protein bound to interleukin 1 (IL-1) responsive element in the IL-6 promoter region was named NF-IL6 (nuclear factor for IL-6). This NF-IL6 was known to be very imporant as a transcription factor for various immuno-protein as well as for IL-6. The human NF-IL6 genes were transfected into the mouse L cells under the metallothionein promoter (MT promoter) to establish a model system for the expression of foreign gene in the mammalian cell line.

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Actin-related protein BAF53 is essential for the formation of replication foci

  • Kwon, Su-Jin;Kwon, Hyock-Man
    • Animal cells and systems
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    • v.16 no.3
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    • pp.183-189
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    • 2012
  • It has been suggested that chromatin is organized into the stable structures that provide fundamental units of chromosome architecture in interphase mammalian cells. The stable structures of chromatin can be visualized as replication foci when replicating DNA is labeled with thymidine analogs. Previously, we showed that the chromosome territory expanded after BAF53 knockdown. In this study, we found that BAF53 is required for the formation of replication foci. DNA replication was not impaired in BAF53 knockdown cells, suggesting that the decrease in the number of replication foci is due to disintegration of replication foci, but not suppression of DNA replication. The attractive forces that maintain structural integrity of replication foci could be disrupted by BAF53 knockdown, and it may be responsible, at least in part, for the expansion of chromosome territories after BAF53 knockdown.

Construction of nervous necrosis virus (NNV) genome-based DNA replicon vectors for the delivery of foreign antigens

  • Jeong In Yang;Ki Hong Kim
    • Journal of fish pathology
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    • v.37 no.1
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    • pp.1-8
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    • 2024
  • The advantages of replicon vectors of RNA viruses include a high ability to stimulate innate immunity and exponential amplification of target mRNA leading to high expression of foreign antigens. The present study aimed to construct a DNA-layered nervous necrosis virus (NNV) replicon vector system in which the capsid protein gene was replaced with a foreign antigen gene and to compare the efficiency of foreign antigen expression between the conventional DNA vaccine vector and the present replicon vector. We presented the first report of a nodavirus DNA replicon-based foreign antigen expression system. Instead of a two-vector system, we devised a one-vector system containing both an NNV RNA-dependent RNA polymerase cassette and a foreign antigen-expressing cassette. This single-vector approach circumvents the issue of low foreign protein expression associated with the low co-transfection efficiency of a two-vector system. Cells transfected with a vector harboring hammerhead ribozyme-fused RNA1 and RNA2 (with the capsid gene ORF replaced with VHSV glycoprotein ORF) exhibited significantly higher transcription of the VHSV glycoprotein gene compared to cells transfected with either a vector without hammerhead ribozyme or a conventional DNA vaccine vector expressing the VHSV glycoprotein. Furthermore, the transcription level of the VHSV glycoprotein in cells transfected with a vector harboring hammerhead ribozyme-fused RNA1 and RNA2 showed a significant increase over time. These results suggest that NNV genome-based DNA replicon vectors have the potential to induce stronger and longer expression of target antigens compared to conventional DNA vaccine vectors.