• 한국균학회지
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• 제47권4호
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• pp.359-371
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• 2019

큰느타리버섯에서 철 저장과 관련된 페리틴 단백질의 발현 및 분비를 최적화하기 위해, T-Fer 벡터에 EcoRI 및 HindIII처리를 해 페리틴 유전자를 얻은 후, BamHI으로 처리된 선형의 pPEVPR1b 분비 벡터에 클로닝하여pPEVPR1b-Fer 재조합 벡터를 구축한 다음 Agrobacterium tumefaciens LBA4404 로 도입하였다. Agrobacterium tumefaciens-mediated transformation 방법에 의해 Pleurotus eryngii로 형질전환하고 kanamycin함유된 MCM 배지에서 올바른 형질전환체를 선별하였고, 단백질 발현은 SDS-PAGE 및 항원항체 반응에 의한 western blot으로 확인하였다. 페리틴 단백질의 분비 발현은 batch culture 및 20 L airlift type fermenter에서 배양 시간 및 온도와 같은 배양 조건에 의해 최적화되었다. 페리틴 생산을 위한 배양 조건은 MCM 배지에서 25℃ 및 8 일 배양에 의해 최적화되었다. 페리틴 단백질의 양은 정량적 단백질 분석에 의해 2.4 mg/g mycelium으로 측정되었다. 그러나, PR1b (32 amino acid)의 분비서열은 큰느타리버섯 내부의 peptidase에 의해 정확하게 processing되지 않았지만, 페리틴 단백질은 균사체에서 최대로 전체단백질의 24.7% 발현되었고, 배양액에서는 검출되지 않았다. 철 결합 활성은 7.5% non-denaturing gel에서 Perls' staining에 의해 확인되었으며, 다량체 페리틴(24 subunits)이 P. eryngii 균사체에서 형성되었음을 보여준다. 생물학적 활성 측정을 위하여 페리틴을 함유한 분말을 제조하여 육계의 사료 첨가제로서의 사용 가능성에 대해 시험하였으며, 결과적으로 페리틴은 육계의 성장을 촉진하고 사료 효율 및 생산 지수를 향상시키는것으로 확인되었다.

• Journal of Applied Biological Chemistry
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• 제50권2호
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• pp.74-77
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• 2007

Crude extracts of 69 marine organisms (27 salt marsh plants and 42 seaweeds) were screened for the inhibitory activity against the protein tyrosine phosphatase 1B (PTP1) in vitro. The most active extracts were methanol extracts from Derbesia marina (80.6% in inhibitory activity) and Symphycladia latiscula (85.6%) at the concentration of $15{\mu}g/mL$. Methanol extracts of Codium adhaerens and Hisikia fuziformis were moderately inhibitory with 71.2 and 69.1% inhibition, respectively. It was peculiar that only the extracts from seaweeds show inhibitory activity where those from salt marsh plants do not show any significant effect.

• Asian Pacific Journal of Cancer Prevention
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• 제15권22호
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• pp.9835-9839
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• 2014

Fisetin is an effective compound extracted from lacquer which has been used in the treatment of various diseases. Preliminary data indicate that it also exerts specific anti-cancer effects. However, the manner in which fisetin regulates cancer growth remains unknown. In this study, we elucidated interference of fisetin with targets of the nuclear factor ${\kappa}B$ signal transduction pathway activated by Epstein-Barr virus encoding latent membrane protein 1 (LMP1)in nasopharyngeal carcinoma (NPC) cells, Results showed that fisetin inhibited the survival rate of CNE-LMP1 cells and NF-${\kappa}B$ activation caused by LMP1. Fisetin also suppressed nuclear translocation of NF-${\kappa}B$ (p65) and $I{\kappa}B{\alpha}$ phosphorylation, while inhibiting CyclinD1, all key targets of the NF-${\kappa}B$ signal transduction pathway. It was suggested that interference effects of fisetin with signal transduction activated by LMP1 encoded by the Epstein-Barr virus may play an important role in its anticancer potential.

• Nutrition Research and Practice
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• 제5권4호
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• pp.288-293
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• 2011

In this study, we investigated the effects of the ethanol extract of aerial parts of Raphanus sativus L. (ERL) on breast cancer cell proliferation and gene expression associated with cell proliferation and apoptosis in MDA-MB-231 human breast cancer cells. The MDA-MB-231 cells were cultured in the presence or absence of various concentrations (100, 200, or 300 ${\mu}g$/mL) of ERL. ERL significantly decreased cell proliferation after 48 h of incubation (P < 0.05). The protein and mRNA expression of $ErbB_2$ were decreased significantly in a dose-dependent manner (P < 0.05). The protein expression of $ErbB_3$ was decreased significantly at an ERL concentration of 300 ${\mu}g$/mL (P < 0.05), and mRNA expression of $ErbB_3$ was decreased significantly in a dose-dependent manner (P < 0.05). The protein expression of Akt was decreased significantly at the ERL concentration of 200 ${\mu}g$/mL (P < 0.05), and the protein expression of pAkt was decreased significantly in a dose-dependent manner (P < 0.05). The mRNA expression of Akt was decreased significantly at the ERL concentration of 200 ${\mu}g$/mL ERL (P < 0.05). The protein and mRNA expression of Bax were increased significantly at ERL concentrations of 200 ${\mu}g$/mL or higher (P < 0.05). The protein expression of $Bcl_2$ was increased significantly at ERL concentrations of 100 ${\mu}g$/mL or higher (P < 0.05), and mRNA expression of $Bcl_2$ was increased significantly at an ERL concentration of 300 ${\mu}g$/mL (P < 0.05). In conclusion, we suggest that Raphanus sativus, L. inhibits cell proliferation via the ErbB-Akt pathway in MDA-MB-231 cells.

• International Journal of Industrial Entomology and Biomaterials
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• 제26권2호
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• pp.119-123
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• 2013

The insect baculovirus expression vector system (BEVS) is useful for producing biologically active recombinant proteins. However, the overexpression of heterologous proteins using this system often results in misfolded proteins and the formation of protein aggregates. To overcome this limitation, we developed a versatile baculovirus expression and secretion system using Bombyx mori protein disulfide isomerase (bPDI) as a fusion partner. bPDI gene fusion was found to improve the secretions and antibacterial activities of recombinant nuecin and enbocin proteins. Thus, we conclude that bPDI gene fusion is a useful addition to BEVS for the large-scale production of bioactive recombinant proteins.

• BMB Reports
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• 제43권7호
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• pp.461-467
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• 2010

Natural products with non-toxic and environmentally friendly properties are good resources for skin-whitening cosmetic agents when compared to artificial synthetic chemicals. Here, we investigated the effect of glyceollin produced to induce disease resistance responses of soybean to specific races of an incompatible pathogen, phytophthora sojae, on melanogenesis and discussed their mechanisms in melanin biosynthesis. We found that glyceollin inhibits melanin synthesis and tyrosinase activity in B16 melanoma cells without cytotoxicity. To elucidate the mechanism of the effect of glyceollin on melanogenesis, we conducted western blot analysis for melanogenic enzymes such as tyrosinase, tyrosinase-related protein-1 (TRP-1), and TRP-2. Glyceollin inhibited tyrosinase and TRP-1 protein expression. Additionally, glyceollin effectively inhibited intracellular cAMP levels in B16 melanoma cells stimulated by $\alpha$-melanocyte stimulating hormone ($\alpha$-MSH). These results suggest that the whitening activity of glyceollin may be due to the inhibition of cAMP involved in the signal pathway of $\alpha$-MSH in B16 melanoma cells.

• 생명과학회지
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• 제25권2호
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• pp.130-135
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• 2015

Pseudoalteromonas elyakovii 유래 alginate lyase 유전자(aly)를 대장균에서 발현시켰을 때, 발현된 대부분의 유전자 산물은 내포체라는 불용성 응집체 형태로 생산되었다. Alginate lyase를 가용성 및 활성형으로 생산하기 위해 Aeropyrum pernix K1 유래의 초고온성 샤페로닌 ApCpnA와 ApCpnB를 공발현 파트너로 도입하였다. aly와 ApCpnA와 ApCpnB 각각과의 공발현 결과, aly 단독발현 때의 alginate lyase 활성 10.1 unit/g-soluble protein에서 ApCpnA와의 공발현 때는 83.1 unit/g-soluble protein, ApCpnB와의 공발현 때는 100.3 unit/g-soluble protein으로 가용성 및 활성형으로의 alginate lyase 생산이 8~10배 크게 향상되었다. 다양한 배양 조건들의 조사를 통해 alginate lyase 최대 생산을 위한 조건은 다음과 같았다: 1.0 mM IPTG, $25^{\circ}C$ 배양 온도(IPTG 유도 후), ApCpnB 공발현 파트너. 이러한 공발현 시스템은 대장균에서 기능적으로 또한 의학적으로 중요한 재조합 단백질의 산업적 생산에 크게 유용하게 사용될 것이다.

• Bulletin of the Korean Chemical Society
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• 제28권6호
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• pp.1005-1009
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• 2007

Leukotriene B4 (LTB4) is a potent chemoattractant for leukocytes and considered to be an inflammatory mediator. Human BLT2 (hBLT2) is a low-affinity G-protein coupled receptor for LTB4 and mediates pertussis toxin-sensitive chemotactic cell movement. Here, we dissected the interaction between hBLT2 and G-protein alpha subunits using GST fusion proteins containing intracellular regions of hBLT2 and various Gα protein including Gα i1, Gα i2, Gα i3, Gα s1, Gα o1, and Gα z. Among the tested Gα subunits, Gα i3 showed the highest binding to the third intracellular loop region of hBLT2 with a dissociation constant (KD) of 5.0 × 10?6 M. These results suggest that Gα i3 has the highest affinity to hBLT2, and the third intracellular loop region of hBLT2 is the major component for the interaction with Gα i3.

• 생물정신의학
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• 제14권3호
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• pp.177-183
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• 2007

목 적: 성상세포(astrocyte)에서 생산되어 신경세포의 증식과분화에 관여하는 S100B 단백이 정신분열병의 진행과 증상론과 관련이 있다는 연구가 계속 진행되고 있다. 이에 정신분열병 환자와 정상대조군의 혈청 S100B 농도를 비교하고, 정신분열병 환자의 증상 양상과 S100B 농도와의 연관성을 연구하였다. 방 법: DSM-IV-TR 진단 기준에 따라 정신분열병으로 진단받은 환자 중 정신분열병 최초 발병 환자 혹은 최소 6개월 간 약물 치료를 하지 않은 정신분열병 환자 21명과 정상대조군 27명의 혈청 S100B 농도를 비교하였으며, 정신분열병 환자의 PANSS 전체점수와 양성증상점수, 음성 증상점수 등과 S100B의 혈청농도 간의 상관관계에 대해서 알아보았다. 결 과: 정신분열병 환자에서의 S100B 혈청농도($0.074{\pm}0.039$ ng/ml)와 정상대조군에서의 S100B 혈청 농도($0.072{\pm}0.030$ng/ml)에는 통계적인 차이는 없었다(p=0.925). 또한, 음성증상점수와 S100B 혈청농도(${\rho}$=0.410, p=0.065 ; 표 3)의 상관관계 및 양성증상점수와 S100B 혈청농도 (${\rho}$=-0.390, p=0.080 ; 표 3)의 상관관계는 통계적으로 유의하지 않았다. 결 론: 이번 연구에서 S100B 혈청농도와 정신분열병과의 관련성을 뚜렷하게 발견하기 어려웠다. 이러한 S100B의 역할과 연관성을 확인하기 위해서 연구대상의 확대와 CSF에서의 농도 측정, 장기적 추적 검사, 다른 관련 물질과의 연관성에 대한 추가적인 연구가 필요하다.

• 농업과학연구
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• 제42권3호
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• pp.237-244
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• 2015

Four ruminally cannulated Holstein steers (BW $401.0{\pm}2.22kg$) fed TMR containing low protein (CP 9.63 %) as a basal diet were used to investigate the effects of cornell net carbohydrates and protein system (CNCPS) fraction enriched protein feeds on rumen fermentation and blood metabolites. The steers used in a $4{\times}4$ Latin square design consumed TMR only (control), TMR with rapeseed meal (AB1), TMR with soybean meal (B2) and TMR with perilla meal (B3C), respectively. The protein feeds were substituted for 30 % crude protein of TMR intake. For measuring ruminal pH, ammonia-N and volatile fatty acids (VFA), ruminal digesta was sampled through ruminal cannula at 1 h-interval after the afternoon feeding. Blood was sampled via the jugular vein after the ruminal digesta sampling. Different CNCPS fraction-enriched proteins did not affect (p>0.05) ruminal pH except B3C being numerically low compared with the other groups. Ammonia-N and VFA were not significantly different among the experimental groups. Numerically low ammonia-N appeared in the steers fed rapeseed meal even though it contained high soluble N composition (A and B1 fractions). The discrepancy is unclear; however this may be related to low protein level in the diet and/or low DM intake. Blood metabolites were not significantly affected by the protein substitution except for blood urea nitrogen that was significantly (p<0.05) increased.