• Title/Summary/Keyword: 30K protein

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A 100 kDa Protein Binding to bHLH Family Consensus Recognition Sequence of RAT p53 Promoter

  • Lee, Min-Hyung;Park, Sun-Hee;Song, Hai-Sun;Lee, Kyung-Hee;Park, Jong-Sang
    • BMB Reports
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    • v.30 no.3
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    • pp.205-210
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    • 1997
  • p53 tumor suppressor plays an important role in the regulation of cellular proliferation. To identify proteins regulating the expression of p53 in rat liver, we analyzed p53 promoter by electrophoretic mobility shift assay (EMSA) and DNase I footprinting assay. We found that a protein binds the sequence CACGTG, bHLH consensus sequence in rat p53 promoter. Southwestern blotting analysis with oligonucleotides containing this sequence shows that the molecular weight of the protein is 100 kDa. This size is not compatible with the bHLH family such as USF or c-Myc/Max which is known to regulate the expression of the human and mouse p53 gene. Therefore this 100 kDa protein may be a new protein regulating basal transcription of rat p53. We purified this 100 kDa protein through sequence-specific DNA affinity chromatogaphy.

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Effects of Fermented Cottonseed and Soybean Meal with Phytase Supplementation on Gossypol Degradation, Phosphorus Availability, and Growth Performance of Olive Flounder (Paralichthys olivaceus)

  • Lim, Se-Jin;Kim, Sung-Sam;Pham, Minh-Anh;Song, Jin-Woo;Cha, Ji-Hoon;Kim, Jin-Dong;Kim, Jung-Un;Lee, Kyeong-Jun
    • Fisheries and Aquatic Sciences
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    • v.13 no.4
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    • pp.284-293
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    • 2010
  • To reduce anti-nutritional factors in plant protein sources for fish meal replacement in fish feeds, cottonseed and soybean meal (CS) were fermented with Aspergillus oryzae. A feeding trial was conducted to verify the effects of fermented CS (FCS) with phytase supplementation on gossypol detoxification, phosphorus digestibility, antioxidant activity, and growth performance of juvenile olive flounder over 10 weeks. Four diets were formulated to replace 0, 30, or 40% fish meal protein with CS or FCS (designated as CS0, CS30, FCS30P, and FCS40P). Phytase (1,000 FTU/kg) was added to FCS30P and FCS40P. The microbial fermentation significantly increased dietary total polyphenols and consequently led to higher DPPH radical-scavenging activities in fish feed and fish tissue. Dietary and liver gossypol concentrations were dramatically decreased by the fermentation process. Phosphorus digestibility was significantly increased in fish fed the FCS40P diet. However, growth performance decreased in fish fed FCS diets. This study demonstrates that the fermentation process and phytase supplementation can improve the phosphorus availability of plant protein sources in fish. The fermentation of CS by A. oryzae could increase antioxidant activities in feed and fish and effectively degrade toxic gossypol in cottonseed meal.

INFLUENCE OF PHENYLALANINE IN THE MEDIUM ON PROTEIN SYNTHESIS OF CHICKEN EMBRYO FIBROBLASTS

  • Kita, K.;Miyazaki, M.;Okumura, J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.9 no.6
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    • pp.701-703
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    • 1996
  • The influence of phenylalanine (Phe) in the medium on protein synthesis of chicken embryo fibroblasts (CEF) was examined. CEF was derived from 9-d-old embryos by trypsin-EDTA digestion. To examine the deficiency of Phe in the medium, CEF was cultured in Dulbecco's modified Eagle's medium (DMEM) with or without Phe. CEF was also cultured in Dulbecco's phosphate buffered saline (PBS ($Ca^{2+}$, $Mg^{2+}$)) with or without $400{\mu}m$ Phe in order to examine the effect of Phe supplementation. All media were supplemented with 10% (v/v) fetal calf serum. After incubation for 6, 30 and 54 h, protein synthesis was measured by the incorporation of L-[2, $6-^{3}H$] Phe into CEF for further 18 h. Protein synthesis of CEF cultured in DMEM was higher than that in PBS ($Ca^{2+}$, $Mg^{2+}$). High specific radioactivity of Phe due to the low concentration of Phe in the medium resulted in the apparent increase in protein synthesis of CEF. Protein synthesis cultured in PBS ($Ca^{2+}$, $Mg^{2+}$) with Phe did not increase during 72 h of cell culture.

Effects of Non-meat Protein Binders and Acidification on the Efficiency of Cold-Set Pork Restructuring by High Pressure

  • Hong, Geun-Pyo;Chun, Ji-Yeon;Lee, Si-Kyung;Choi, Mi-Jung
    • Food Science of Animal Resources
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    • v.32 no.3
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    • pp.301-307
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    • 2012
  • We investigated the effects of non-meat protein binders combined with glucono-${\delta}$-lactone (GdL) on the binding properties regarding restructured pork prepared by high-pressure treatment. Soy protein isolate (SPI), casein (CS), whey protein concentrate (WPC), and egg white (EW) were used as non-meat protein binders and compared with the control (no binder) and with the ${\kappa}$-carrageenan (KC) treatment. The compression and depression rates were 2.3 and 37 MPa/s, respectively, and pressurization was conducted at 200 MPa for 30 min at $4^{\circ}C$. After pressurization, the physical properties (pH, water-holding capacity, color, tensile strength, and microscopic structure) of the sample were evaluated. The combination of pressurization with acidification enabled cold-set meat binding, and the binding strength of restructured pork was enhanced by the addition of non-meat proteins. Among binders, SPI demonstrated the best efficiency in binding meat pieces. Therefore, the present study demonstrated that the combination of acidification and pressurization processes with the utilization of non-meat protein binders has a potential benefit in meat restructuring.

Purification Properties and Treatment Conditions of Cellulase for Papermaking Processes (제지 공정 적용을 위한 Cellulase의 정제 특성 및 처리 조건 선정에 관한 연구)

  • Kim, Byung-Hyon;Jeon, Yang
    • Journal of Korea Technical Association of The Pulp and Paper Industry
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    • v.30 no.1
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    • pp.18-28
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    • 1998
  • The main protein peak was observed in fraction No.9 and 109 when cellulase seperation was conducted by use of DEAE-Sephadex. The protein obtained from fraction No.9 has the characteristics of Cx component and that from fraction No.109 characteristics similiar to $C_1$ component. The effective reaction condition of the ensyme used was $40^{\circ}C$ in temperature. pH 5.0 and 90 minutes in treatment time. For the case of $C_1$ pH 5.5 in temperature range of $30^{\circ}C 50^^{\circ}C$, 4.0 5.5 in pH, and over 30 minutes of treatment time, the reaction was in the range of 80% of the maximum. Affinity of enzyme increased as freeness, increased, and this effect was more visible in fiber than in fines.

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The Use of Meat Meal as a Dietary Protein Source Replacing Fish Meal in Juvenile Rockfish Sebastes schlegeli

  • Lee, Yong-Whan;Lee, Sang-Min
    • Journal of Aquaculture
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    • v.18 no.2
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    • pp.92-97
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    • 2005
  • This study examined the partial replacement of the fish meal with meat meal in practical diets for juvenile rock-fish. Five isonitrogenous (48% CP) diets were prepared to contain meat meal at 0% (control), 10%, 20%, 30% and 40% with substituting the mackerel meal in the control diet. Three replicate groups of fish (initial average weight, 4.1g) were hand-fed to visual satiety two times daily for 8 weeks. Survival (>93%) and daily feed intake were not significantly different (P>0.05) among treatments. The best weight gain, feed efficiency and protein efficiency ratio were obtained from fish fed the diets containing 0% and 10% meat meal, and were not significantly different (P>0.05) to those of fish 134 diet containing 20% meat meal. Condition factor, visceralsomatic index and hepatosomatic index were not influenced by dietary meat meal levels. The contents of crude protein and ash of whole body were not significantly affected (P>0.05) by dietary meat meal levels, whereas crude lipid content of fish fed the diets containing 30% and 40% was lower than that of fish fed the control diet. Proximate composition of liver was not influenced by dietary meat meal level (P>0.05). The data obtained in this study indicate that a diet containing $10{\sim}20%$ meat meal could be used for least-cost formulation in juvenile rockfish diet.

Development of Hydrogel for Decrease Protein Adsorption and Application to Intraocular Lens (단백질흡착 감소용 하이드로겔의 개발 및 인공수정체로의 적용)

  • Ko, Na Young;Lee, Hyun Mee
    • Journal of the Korean Chemical Society
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    • v.62 no.1
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    • pp.30-35
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    • 2018
  • The purpose of this study was to decrease the protein adsorption and improve the function of the hydrophobic acrylic Intraocular lens(IOL). Hydrophobic acrylic intraocular lenses were prepared by using ethyleneglycol phenyletheracrylate (EGPEA), styrene and 2-hydroxyethyl methacrylate (HEMA). Polyvinyl pyrrolidone (PVP) and 2-methacryloyloxyethyl phosphorylcholine (MPC) were used as additives. Water contents, wettability, light transmittance and protein adsorption amount were measured to evaluate the physical properties of the intraocular lens. The water content and wettability of all samples containing additives were increased and the amount of protein adsorption decreased. In particular, samples containing MPC showed a further decrease in protein adsorption. The hydrophobic acrylic intraocular lens with PVP and MPC was found to improve the function of the intraocular lens by reducing the protein adsorption while having basic physical properties.

Influence of Adding Recovered Protein from Processing Wastewater on the Quality of Mechanically Separated Chicken Meat Surimi Like-Material

  • Cortez-Vega, William Renzo;Fonseca, Gustavo Graciano;Bagatini, Daniela Cardozo;Prentice, Carlos
    • Food Science of Animal Resources
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    • v.37 no.2
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    • pp.162-167
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    • 2017
  • Functional and nutritional soluble proteins can be recovered from surimi (and surimi-like material) processing wastewater, reducing environmental problems and the cost of an irresponsible waste disposal. Recovered proteins may be added back at a low percentage to surimi products. The aim of this work was to evaluate the effect of the addition of soluble recovered proteins (RP), obtained from mechanically separated chicken meat surimi-like material (MSCM-SLM) processing wastewater by acidic pH-shifting, on the composition and texture of RP-MSCM-SLM, with RP contents of 0, 10, 20 and 30% (w/w) in the mixture. For that, proximate composition and gel properties were evaluated. The fat content of the MSCM-SLM was significantly reduced to 11.98% and protein increased to 83.64% (dry basis) after three washing cycles. The addition of 30% RP in the MSCM-SLM significantly augmented the protein content to 93.45% and reduced fat content from to 2.78%. On the other hand, the addition of RP was responsible for a drastic decrease in texture parameters, reaching 252.36 g, 185.23 g.cm, and 6.97 N for breaking force, gel strength and cutting strength, respectively, when 30% of RP was included in the MSCM-SLM. It was concluded that the obtained intermediary product (RP-MSCM-SLM) is a good option to applications in processed meat products where high texture parameters are dispensable, e.g., emulsified inlaid frankfurter-type sausages, but high protein content and low fat content desired.

Enhanced In Vitro Protein Synthesis Through Optimal Design of PCR Primers

  • Ahn Jin-Ho;Son Jeong-Mi;Hwang Mi-Yeon;Kim Tae-Wan;Park Chang-Kil;Choi Cha-Yong;Kim Dong-Myung
    • Journal of Microbiology and Biotechnology
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    • v.16 no.3
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    • pp.355-359
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    • 2006
  • The functional stability of mRNA is one of the crucial factors affecting the efficiency of in vitro translation. As the rapid degradation of mRNA in the cell extract (S30 extract) causes early termination of the translational reactions, extending the mRNA half-life will improve the productivity of the in vitro protein synthesis. Thus, a simple PCR-based method is introduced to increase the stability of mRNA in an S30 extract. The target genes are PCR-amplified with primers designed to make the ends of the transcribed mRNA molecule anneal to each other. When compared with normal mRNA, the mRNA with the annealing sequences resulted in an approximately 2-fold increase of protein synthesis in an in vitro translation reaction. In addition, sequential transcription and translation reactions in a single tube enabled direct protein expression from the PCR-amplified genes without any separate purification of the mRNA.