• Title/Summary/Keyword: $P_{tac}$

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PelB Signal Sequence로 유도된 재조합 인간 상피세포 증식인자 분비 발현 벡터의 제조

  • Park, Se-Cheol;Nam, Jung-Hyun;Kim, Jeong-Keun;Kwon, Tae-Jong;Ko, In-Young;You, Kwang-Hyun
    • Microbiology and Biotechnology Letters
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    • v.24 no.5
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    • pp.553-559
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    • 1996
  • We have designed nucleotide sequences of hEGF structural gene to eliminate the N-terminal methionine residue incorporated during the translation initiation step, and constructed recombinant human epidermal growth factor (rhEGF) secretion plasmids pYHB101, and pYHB2 in which pelB signal sequence-hEGF gene was expressed under the control of the T7, and tac promoter, respectively. We also constructed pYHB1 vector which contains rhEGF gene controlled by T7 promoter. The transformant with pYHB101 showed relatively slow growth pattern compared to the transformant with pYHB1. However, we observed that the transformant with pYHB101 secreted rhEGF of 13 mg/l significantly after 5 hr induction with 1 mM IPTG and that the T7 promoter was more effective than tac promoter when connected to pelB signal sequence. The amount of rhEGF was 14 mg/l under the sub-optimized condition.

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Cloning and Expression of Human Immunodeficiency Virus-1 Epitopes in Escherichia coli (대장균에서 사람의 면역결핍 바이러스-1 epitopes 클로닝과 발현에 대한 연구)

  • 유향숙;장원희;박희동;현상원;남상욱;이영익
    • Korean Journal of Microbiology
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    • v.29 no.1
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    • pp.1-7
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    • 1991
  • Human immunodeficiency virus type 1 (HIV-1) causes a deadly infectious disease, Acquired Immunodeficiency Syndrome (ADIS). As a first step to develop a reliable and fast diagnostic procedure for HIV-1 infection, we cloned various immunodominant epitopes of HIV-1 in bacterial expression vectors containing tac or trp promoter. While the protein level of direct expression of gp160 was low, trp E fused gp120, gp41 and p17-p24 were produced at high levels (15-30% of total bacterial proteins) in E. coli. Since gp120 and gp41 contain relatively conserved regions which can react with antibodies in the plasma from most of HIV-1 infected individuals, these expression clones were used for large preparations of HIV-1 antigens.

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Vitamin C and antioxidant capacity stability in cherry and romaine during storage at different temperatures (저장 조건에 따른 로메인 및 체리의 비타민 C 함량 및 항산화능의 변화)

  • Park, Hee Jung;Lee, Myung Joo;Lee, Hye Ran
    • Journal of Nutrition and Health
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    • v.49 no.1
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    • pp.51-58
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    • 2016
  • Purpose: The aim of this work was to study the change in antioxidant activity depending on storage temperature and storage period in romaine and cherry. Method: The plant material was stored at $0.7{\pm}0.6^{\circ}C$, $3.5{\pm}2.8^{\circ}C$, and $4.7{\pm}1.4^{\circ}C$. Cherry and romaine were stored for a period of 9 days and 7 days, respectively. The cherry was taken from each group of samples at regular intervals of days and the romaine was taken from each group of samples at regular intervals of 2days. Vitamin C, total polyphenol, and total flavonoid stability and antioxidant capacity including DPPH, total antioxidant capacity (TAC) were measured. Results: For cherry, the levels of TAC and flavonoid were higher at the $0.7{\pm}0.6^{\circ}C$ condition than other conditions (p < 0.05). The polyphenol and vitamin C levels were not significantly different among storage conditions. In the case of romaine, the level of TAC was highly preserved until 7 days at the $0.7{\pm}0.6^{\circ}C$ condition. Vitamin C level was significantly lower at the $3.5{\pm}2.8^{\circ}C$ condition (p < 0.05). DPPH activity was highest at the $0.7{\pm}0.6^{\circ}C$ condition (p < 0.05). DPPH activity was shown in order of $0.7{\pm}0.6^{\circ}C>4.7{\pm}1.4^{\circ}C>3.5{\pm}2.8^{\circ}C$. Conclusion: The results indicated that the narrow differences and fluctuation in temperature were associated with antioxidant capacity and it might enhance the nutritional shelf life of vegetables and fruits.

Studies on the Separation and Preconcentration of Metal Ions by Chelating Resin containing (Polystyrene-divinylbenzene)-thiazolylazo Phenol Derivatives(I) ((Polystyrene-divinylbenzene)-thiazolylazo phenol형 킬레이트 수지에 의한 금속이온의 분리 및 농축에 관한 연구(I))

  • Lim, Jae-Hee;Kim, Min-Kyun;Lee, Chang-Hun;Lee, Won
    • Analytical Science and Technology
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    • v.9 no.3
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    • pp.279-291
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    • 1996
  • The new chelating resins, XAD-2, 4, 16-TAC and XAD-2, 4, 16-TAO were synthesized by Amberlite XAD-2, XAD-4, and XAD-16 macroreticular resins with 2-(2-thiazolylazo)-p-cresol(TAC) and 4-(2-thiazolylazo)orcinol(TAO) as functional groups and were characterized by elemental analysis and FT-IR spectrometry. It was found that the content of functional group in chelating resin was 0.60mmol/g in XAD-16-TAC and 0.68mmol/g in XAD-16-TAO respectively. The chelating resins were stable in acidic and alkaline solution and can be reused over 10 times. The sorption behavior of some metalions to two chelating resins was investigated by batch method, which included batch equilibrium, effect of pH, coexisting ions and masking agent. For the optimum condition of sorption, the time required for equilibrium was about 1 hour and optimum pH was 5. In the presence of anions such as ${SO_4}^{2-}$ and $CH_3COO^-$, the sorption of U(VI) ion was slightly reduced but other anions such as $Cl^-$ and $NO{_3}^-$ revealed no interference effect. Also, sorption capacity of U(VI) ion was decreased by addition of $CO{_3}^{2-}$ ion because of complex formation of $[UO_2(CO_3)_3]^{4-}$, but alkali metals and alkali earth metals including Na(I), K(I), Mg(II), and Ca(II) were not affected for the sorption extent. Masking agent, NTA showed better separation efficiency of U(VI) ion from coexisting metal ions such as Th(IV), Zr(IV), Hf(IV), Cu(II), Cd(II), Pb(II), Ni(II), Zn(II) and Mn(II) than EDTA, CDTA.

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Association of total dietary antioxidant capacity with oxidative stress and metabolic markers among patients with metabolic syndrome (대사증후군 환자 및 위험군의 식사 내 총 항산화능과 산화스트레스 및 대사 지표의 연관성)

  • Ham, Dongwoo;Jun, Shinyoung;Kang, Minji;Shin, Sangah;Wie, Gyung-Ah;Baik, Hyun Wook;Joung, Hyojee
    • Journal of Nutrition and Health
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    • v.50 no.3
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    • pp.246-256
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    • 2017
  • Purpose: This study aimed to investigate the association of total dietary antioxidant capacity (TAC) with oxidative stress and metabolic markers among patients with metabolic syndrome according to gender. Methods: A total of 346 subjects aged 30~59 years with two or more risk factors of metabolic syndrome were recruited from a general hospital near Seoul in South Korea between 2010 and 2012 based on data from the medical checkup. Biochemical indices for oxidative stress and metabolic markers were measured. Food consumption data from 3-day food records were linked with the antioxidant capacity database for commonly consumed Korean foods to estimate individual's TAC. Results: Average dietary TAC of the study subjects was 132.0 mg VCE/d/1,000 kcal in men and 196.4 mg VCE/d/1,000 kcal in women. Levels of ${\gamma}$-glutamyltransferase (GGT), systolic blood pressure, diastolic blood pressure, and blood triglycerides were reduced significantly according to increasing TAC in men, but there was no significant trend in women. Intakes of total flavonoids and carotenoids were significantly negatively correlated with GGT (p < 0.05) and d-ROMs (p < 0.01) in men, whereas those of ${\alpha}$-tocopherol (p < 0.05) and ${\gamma}$-tocopherol (p < 0.05) were positively correlated with biological antioxidant potential (BAP) in women. The odds ratio of high oxidative stress indices and abnormal metabolic markers according to TAC level were not significant in either men or women. Conclusion: The results show that dietary TAC was partially associated with oxidative stress and metabolic markers among patients with metabolic syndrome. Further research is required for elucidating the association between dietary TAC and incidence of metabolic syndrome and chronic diseases within a large population in prospective studies.

Derepression of a Methionine Biosynthetic Gene by Utilizing a Promoter Isolated from Corynebacterium glutamicum (Corynebacterium glutamicum에서 분리된 프로모터를 이용한 메치오닌 생합성 유전자의 조절해제)

  • Park Soo-Dong;Park Ik-Hyun;Choi Jong-Soo;Kim Il-Kwon;Kim Younhee;Lee Heung-Shick
    • Korean Journal of Microbiology
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    • v.41 no.4
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    • pp.300-305
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    • 2005
  • A transcriptionally active fragment $(P_{19})$ isolated by utilizing the promoter-probe shuttle vector pSK1Cat was analyzed. By subcloning analysis, the 180 bp region $(P_{180})$ responsible for the activity was determined. Transcriptional fusion of the C. glutamicum metX gene to $P_{180}\;(P_{180}-metX)$ resulted in a 24-fold increase in MetX activity in a complex medium, while a 13-fold increase was observed with the $P_{tac}$ promoter. Additionally, the expression conferred by $P_{180}$ was not affected by methionine added to the growth medium, suggesting that the $P_{180}$ clone is useful for the deregulated expression of biosynthetic genes in C. glutamicum during amino acid fermentation. Introduction of $P_{180}-metX$ into a lysine-producing C. glutamicum resulted in the production of methionine to 0.8 g/l.

Subcloning and Enhanced Expression of the $\beta$-Xylosidase Gene Cloned from Alkalophilic Bacillus sp. K-17 (호알칼리성 Bacillus sp. K-17 의 $\beta$-Xylosidase 유전자의 Subcloning 및 발현증진)

  • Sung, Nack-Kie;Ko, Hack-Ryong;Kho, Yung-Hee;Chun, Hyo-Kon;Chung, Young-Chul
    • Microbiology and Biotechnology Letters
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    • v.17 no.4
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    • pp.283-288
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    • 1989
  • To reduce the size of 5.0kb HindIII fragment containing $\beta$-xylosidase gene, the 5.0kb insert of pAX278 which was previously cloned was reduced by various deletions and thus 1.4kb EcoRI-Xbal fragment was subcloned into pUC19, and the recombinant plasmid was named pAK208. The $\beta$-xylosidase acnivity of E. coli harboring pAK208 was higher about 1.3times than that of pAX278. For the improvement of $\beta$-xylosidase activity, we cloned and expressed the $\beta$-xylosidase gene in E. coli using vector pKK223-3 containing a potent tac-promoter, and enzyme activity of the transformant harboring pKHR212 was increased about 3.3 and 1.8 times than that of E. coli(pAX278) and Bacillus sp. K-17, respectively. To obtain better expression of $\beta$-xylosidase gene, the whole 5.0kb HineIII fragment was recloned into pC194, and the Bacillus sp. K-17 transformant harbor-ing the recombinant plasmid pCX174 showed higher activity than that of the E. coli (pAX278) and Bacillus sp. K-17, respectively. The characteristics of enzyme purified from transformants were consistent with those front alkalophilic Bacillus sp, K-17.

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Effective Treatment of Swine Manure with Chinese Cabbage Silage through Two Serial Anaerobic Digestion (돈분과 배추사일리지를 이용한 2조 혐기소화의 효과적인 처리)

  • Kim, Sang-Hun;Kafle, Gopi Krishna
    • Journal of Biosystems Engineering
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    • v.35 no.1
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    • pp.53-63
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    • 2010
  • The performance of two serial completely stirred tank reactors (CSTRs) for treatment of swine manure (SM) with Chinese cabbage silage (CCS) was studied. The two CSTRs of 41 liters each were connected in series and first reactor was fed swine manure with Chinese cabbage silage in 3:1 proportion by VS basis. The FOS/TAC, methane content (%) and pH were utilized as the parameters for the digester stability control. The FOS/TAC value was found to be effective indicator for instant digester condition. The stability of reactors can be obtained with the FOS/TAC value up to 1.0 with accumulation of FOS value below 10,000 mg/L. Material exchange method was effective in transferring the excess volatile fatty acids (VFA) from the first reactor to the second one and maintaining stability in both the reactors. The biogas yield and the methane yield was 0.55-0.61 and 0.41-0.42 L/g VS fed, respectively, at organic loading rate (OLR) of 2.2-2.6 g VS/L with total HRT of 32 days.

A Stock Assessment of Yellow Croaker using Bioeconomic Model: a Case of Single Species and Multiple Fisheries (생물경제모형을 이용한 참조기의 자원평가에 관한 연구 - 단일어종·다수어업 사례를 중심으로)

  • Sim, Seonghyun;Nam, Jongoh
    • Ocean and Polar Research
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    • v.37 no.2
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    • pp.161-177
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    • 2015
  • This study analyzes the stock assessment of yellow croaker caught mainly by the Korean stow net and gill net fisheries focusing on single species and multiple fisheries. This study standardizes fishing efforts for the two fisheries using the general linear model and uses a surplus production model based on the exponential growth model. The Clarke Yoshimoto Pooley model estimates a maximum sustainable yield(MSY), an allowable biological catch(ABC), fishing efforts for MSY($E_{MSY}$) and for ABC($E_{ABC}$). The bio-economic model is used to estimate the maximum economic yield(MEY) and fishing efforts for MEY($E_{MSY}$). Also, the study employs an economic analysis to estimate the economic interaction between stow net and gill net fisheries. The economic analysis shows the profit accruing to the two fisheries from estimated ABC. Finally, the study compares TACs based on single species and single fishery to TAC based on single species and multiple fisheries. The study proposes that the TAC assessment is necessary for single species and multiple fisheries in order to preserve resources.

Interactions between Oxidative Stress, Lipid Profile and Antioxidants in Breast Cancer: A Case Control Study

  • Gupta, Rakesh Kumar;Patel, Amit Kumar;Kumari, Rajni;Chugh, Seema;Shrivastav, Chitrangada;Mehra, Siddharth;Sharma, Ajay Narayan
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.12
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    • pp.6295-6298
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    • 2012
  • Oxidant/antioxidant balance has been suggested as an important factor for initiation and progression of cancer. The objective of this study was to determine changes in the levels of malondialdehyde (MDA), nitric oxide (NO), total cholesterol, triglycerides, LDL-cholesterol, HDL-cholesterol, total antioxidant capacity (TAC), glutathione peroxidase (GSH-Px), and superoxide dismutase (SOD) activities in serum samples of breast cancer patients (n=30) and healthy subjects (n=100). MDA and NO levels were found to be increased in breast cancer patients compared to the healthy subject group (p<0.05). Total cholesterol and triglycerides were elevated; and HDL-cholesterol level was found to be decreased in the cancer patients as compared to the healthy subjects (p<0.05). Compared to the healthy group, both serum TAC levels (p<0.001) and activity of SOD and GSH-Px (p=0.05) were found to be decreased in the breast cancer patients as compared to the healthy controls. Considering the data presented in this study, we suggest that free radicals induce lipid eroxidation and peroxidation of unsaturated fatty acid with decreased activity of enzymatic antioxidants in breast cancer.