• Title/Summary/Keyword: incubation time and temperature

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Studies on the Factors Influencing the Transformation in Escherichia with pBR322 DNA (Escherichia coli의 pBR322 DNA 형질전환에 관여하는 인자에 관한 연구)

  • Yoo, Han-sang;Mah, Jum-sool
    • Korean Journal of Veterinary Research
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    • v.24 no.1
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    • pp.40-49
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    • 1984
  • To investigate the factors influencing the artifical transformation in Escherichia coli, E. coli C600 was transformed by pBR322 DNA with tetracycline and ampicillin resistant gene purified by CsCl-Etbr equilibrium density gradient centrifugation from E.coli HB 101. The influencing factors in the transformation such as concentration of calcium chloride, time of ice incubation, temperature and time of heat shock, time of gene expression, effects of plasmid DNA concentration and adding time were examined in these experiments. The results obtained were as follows; 1. The highest transformation frequency was observed in the treatments of 100 mM $CaCl_2$ before heat shock and the treatment of $CaCl_2$ was essential step in the process of E. coli transformation. 2. The highest transformation frequency was observed in the treatment of heat shock at $42^{\circ}C$ for 4 min. or $37^{\circ}C$ for 6 min., but the prolonged heat shock resulted a decreased transformation frequency. 3. Treatments of ice incubation at $0^{\circ}C$ for 45 min. before heat stocks or at $0^{\circ}C$ for 30min. after heat shock resulted an increased transformation frequency. 4. There was a linear relationship between DNA concentration and transformation frequency at the concentration of $8{\times}10^3$ recipient cells. The highest transformation frequency reached in carte of 7 mcg of donor DNA, but above 1 mcg of DNA concentration, transformation frequency was not remarkably increased. Addition of donor DNA just after the treatment of $CaCl_2$ was the best. 5. The best condition of gene expression at $37^{\circ}C$ were 40min. for TC-resistant gene and 100min. for AP-resistant gene. TC-resistant gene was higher in the transformation frequency and faster in the gene expression time than AP-resistant gene. In these results, the best conditions for the transformation of E. coli C 600 with pBR322 DNA were: treatment with 100mM $CaCl_2$, ice incubation at $0^{\circ}C$ for 45 min, heat shock at $42^{\circ}C$ for 4 min., 30 min. of ice incubation and incubation at $37^{\circ}C$ for 100min. for gene expression in that order.

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Improvement of Occasional Artificial Hatching and Incubation Method in Diapause Egg of the Wild Silkmoth, Antheraea yamamai (천잠알의 수시부화 및 최청법 개선)

  • Yoon, Hyung-Joo;Kang, Pil-Don;Lee, Sang-Mong;Kim, Sam-Eun;Kim, Ki-Young
    • Korean journal of applied entomology
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    • v.46 no.2
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    • pp.295-302
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    • 2007
  • We investigated starvation of hatching larvae, occasional artificial hatching and incubation method to establish year-round rearing of the wild silkmoth, Antheraea yamamai. In the test of starvation of hatching larvae for brushing at a time, the survival rate of the fourth instar of larvae starved for 1 day after hatching in $25^{\circ}C\;and\;5^{\circ}C$ was 83.3% and 96.0%, respectively. The result represents that the survival rate is high at low temperature during starvation. In the occasional artificial hatching test for multi-times rearing of A. yamamai, the useful hatchability is high at $5^{\circ}C$ in case of preserving eggs for 2 months from incubation time, and at both $2.5^{\circ}C\;and\;0^{\circ}C$ in case of over 6 months. A new incubation method with pre-incubation at $15^{\circ}C$ and 24 D photoperiod showed high hatchability about 80% for only 2 days compared with hatching for 5-6 days in traditional incubation method with the preservation at $25^{\circ}C$.

The Novel Assay Method for Thrombin by Weighing Fibrin Clot (피브린의 무게측정에 의한 새로운 트롬빈활성측정법)

  • Park, Inshik;Kim, Gi-Nahm
    • Journal of Life Science
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    • v.14 no.3
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    • pp.441-444
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    • 2004
  • This study was performed to establish a simple and rapid method for measuring thrombin activity based on weight of fibrin clot formed. The new method was based on the weight measurement of fibrin clot after enzymatic reaction of thrombin with fibrinogen. The fibrin formation depended upon the activities of thrombin used, temperature, incubation time, and centrifugation time. The fibrin formation was increased proportionally up to 1.0 unit/ml of thrombin activity, 4.0mg/ml of fibrinogen concentration, and 5 min of incubation time at 37$^{\circ}C$. The fibrin clot formed was stable by centrifugation at 3,000$\times$g for 5min. This simple assay based on weight of fibrin after centrifugation would be useful for identifying natural food anticoagulants by inhibiting thrombin.

Influence of Culture Comditions on Acid Tolerance of Lactobacillus casei YIT 9018 (배양조건에 따른 Lactobacillus casei YIT 9018의 니산성 변화)

  • Sim, Jae-Hun;Kim, Sang-Kyo;Baek, Young-Jin;Oh, Tae-Kwang;Yang, Han-Chul
    • Microbiology and Biotechnology Letters
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    • v.23 no.1
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    • pp.17-23
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    • 1995
  • We studied the influence of culture conditions on the acid tolerance of Lactobacillus casei YIT 9018 in artificial gastric juice with respect to relative amount of membrane bound ATPase and their biochemical characteristics. With raising incubation temperature from 30.5$\circ$C to 40.5$\circ$C and lengthening incubation time from exponential phase to late stationary phase, the acid tolerance of L. casei YIT 9018 was increased. As acid tolerance enhanced, C$_{18:1}$ content of membrane fatty acid was reduced and C$_{19:0 cyclo}$ was enriched but the others were not changed greatly. At high ATPase activity, proton permeability was relatively low but this phenomenon did not correspond to acid tolerance. In conclusion, it was considered that changes of C$_{18:1}$ and C$_{19:0 cyclo}$ were closely related to the acid tolerance of L. casei YIT 9018.

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Optimal Conditions for the Production of Antioxidant by Nocardiopsis sp. S-1

  • Moon, Young-Gun;Kim, Man-Chul;Heo, Moon-Soo
    • 한국생물공학회:학술대회논문집
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    • 2005.10a
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    • pp.364-367
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    • 2005
  • This study investigated the production of antioxidant from Actinomyces culture supernatant. For the research of the natural marine antioxidant, several bacteria were isolated from the coast of Je-ju in Korea. An actinomycetes strains, S-1 was identified to a genus level 16S ribosomal DNA sequence and fatty acid analysis. From these results and other characteristics described in the Bergey's Manual, this strain was identificated as a Nocardiopsis dassonvillei. Strain S-1 showed high activity of 1,1-diphenyl-2-prcrylhydrazyl radical scavenging. The hydroxyl radical scavenging ability of Nocardiopsis sp. S-1 supernatant was 53%. Nutritional and cultural conditions for the production of antioxidant by this organism under shake-flask conditions have optimized. Similary initial medium pH 7.6, incubation temperature of $25^{\cicr}C$, sodium chloride concentration 2.5 and incubation time of 8 day were found to be optimal.

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Effects of Aging Temperature and Time on the Conversion of Garlic (Allium sativum L.) Components (온도 및 숙성기간이 마늘의 화학적 성분변화에 미치는 영향)

  • Cho, Kang-Jin;Cha, Ji-Young;Yim, Joo-Hyuk;Kim, Jae-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.1
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    • pp.84-88
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    • 2011
  • Some thermally processed foods have higher biological activities due to their various chemical changes during heat treatment. Especially, 5-hydroxymethylfurfural (HMF) is derived from dehydration of sugars and has been identified in processed garlic. The biological function of HMF have revealed as antisickling agent and thyrosinase inhibitor. This study was carried out to examine the formation of HMF and free sugars from the aged garlics when it is treated at 60 and $75^{\circ}C$ and different incubation periods from 7 to 35 days. HMF and free sugars from the hot-water extracts of aged garlics were analyzed with GC/MS, LC/MS, and HPLC. The amount of HMF was higher than at $75^{\circ}C$ and increasing incubation period. Among free sugars, the only fructose except glucose and sucrose was formed and converted to HMF at high temperature and long incubation period. However, fructose formed in low temperature during making of aged garlic was rarely converted to HMF. This result indicates that formation of HMF can be dependent on the temperature and incubation period for making aged garlic.

Dissolution and Precipitation behaviors of Complex Carbonitrides in Austenite of a V-Nb Microalloyed Steel (V-Nb Microalloyed 강의 오스테나이트역에서 복합 탄질화물의 재용해 및 석출 거동)

  • Ha, Yangsoo;Jung, Jae-Gil;Lee, Young-Kook
    • Korean Journal of Metals and Materials
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    • v.49 no.12
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    • pp.917-923
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    • 2011
  • Dissolution and precipitation behaviors of complex carbonitrides in austenite of a V-Nb microalloyed steel were quantitatively examined through electrical resistivity measurement and transmission electron microscopy. The electrical resistivity increased with solution treatment temperature up to $1240^{\circ}C$ and then was saturated at $225n{\Omega}m$ for a holding time of 10 min. The electrical resistivity method was also used to quantitatively measure the isothermal precipitation kinetics of the complex carbonitrides in austenite. Nb-rich precipitates were observed in austenite at the early stages of precipitation, but Nb was replaced by V up to the equilibrium amount within the precipitates with further holding time. The time-temperature-precipitation diagram showed a C-type curve; nose temperature and its incubation time were $900^{\circ}C$ and 100 s, respectively.

The Effects of Different Holding Temperatures and Storage Time during the Pre-Incubation Period on the Hatchability of Hens Eggs in Broiler Bleeders (육용종계 종란의 보관온도에 따른 입란 전 저장기간이 부화율에 미치는 영향)

  • 강보석;서옥석;나재천;김상호;김학규;장병귀;김태호;이상진;하정기
    • Korean Journal of Poultry Science
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    • v.29 no.2
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    • pp.89-94
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    • 2002
  • This study was performed to evaluate effects of different holding temperatures and storing periods during the pre-incubation period on egg hatchability of hens egg in broiler breeders. For the treatments 1(T1)~7(T7), which were stored fur 1(T1) to 7 days(T7) before egg incubation, respectively. There were three replicates per treatment and forty eggs per replicate. This study was performed twice, which were 40(Summer) and 50 weeks of age(Autumn) in broiler breeders. Storing ambient temperature of egg, egg weight, at 0 and 18 days during incubation, fertility, hatchability and embryo mortality were examined. Average hatchability was rapidly decreased only in Summer. Although it was not significantly different in Autumn. This experiment was concluded that storing periods of hatchery egg was influenced hatchability, especially in high ambient temperature conditions(Summer, above $25^{\circ}C$ ). In conclusion, we found out that optimum hatchability can be achieved with a storage temperature of 13 ~$19^{\circ}C$ for broiler breeder eggs stored for up to 7 days.

Effect of Maillard reaction with xylose, yeast extract and methionine on volatile components and potent odorants of tuna viscera hydrolysate

  • Sumitra Boonbumrung;Nantipa Pansawat;Pramvadee Tepwong;Juta Mookdasanit
    • Fisheries and Aquatic Sciences
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    • v.26 no.6
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    • pp.393-405
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    • 2023
  • The aim of this research was to enhance the flavor of visceral extracts from skipjack tuna. Flavor precursors and the optimum condition for the Maillard reaction were determined. The flavor extract was prepared from the tuna viscera using Endo/Exo Protease controlled in 3 factors; temperature, enzyme amounts and incubation time. The optimal condition for producing tuna viscera protein hydrolysate (TVPH) was 60℃, 0.5% enzyme (w/w) and 4-hour incubation time. TVPH were further processed to tuna viscera flavor enhancer (TVFE) with Maillard reaction. The Maillard reactions of TVFE were conducted with or without supplements such as xylose, yeast extract and methionine. The Maillard volatile components were analyzed with gas chromatography-mass spectrometry. Sixteen volatiles such as 2-methylpropanal, methylpyrazine, 2,5-dimethylpyrazine, dimethyl disulfide and 2-acetylthaizone were newly formed via Maillard reaction and the similarity of volatile contents from TVPH and TVFE were virtualized using Pearson's correlation integrated with heat-map and principal component analysis. To virtualize aromagram of TVPH and TVFE, odor activity value and odor impact spectrum (OIS) techniques were applied. According to OIS results, 3-methylbutanal, 2-methylbutanal, 1-octen-3-ol 2,5-dimethylpyrazine, methional and dimethyl trisulfide were the potent odorants contributed to the meaty, creamy, and toasted aroma in TVFE.

Optimization for the Production of Mono- and Di-acylglycerols from Corn Oil by Enzymic Glycerolysis Using Response Surface Methodology (반응표면분석에 의한 옥수수유 유래 monoacylglycerol과 diacylglycerol 합성 조건의 최적화)

  • Park, Rae-Kyun;Choi, Sang-Won;Lee, Ki-Teak
    • Korean Journal of Food Science and Technology
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    • v.36 no.5
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    • pp.717-722
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    • 2004
  • Response surface methodology was used to optimize production conditions of monoacylglycerol (MAG) and diacylglycerols (DAG) from corn oil by enzymic glycerolysis. Contents of $1,3-DAG\;(Y_1),\;1,2-DAG\;(Y_2),\;total\;DAG\;(Y_3),\;MAG\;(Y_4)$, and total $DAG+MAG\;(Y_5)$ were obtained. Conditions were optimized using central composite design with incubation temperature $(35-75^{\circ}C,\;X_1)$, incubation time (1-11 hr, $X_2$), and amount of hexane added (0-2 mL, $X_3$) as three variables. Content of 1,3-DAG was maximized by 20.43 area% at incubation temperature of $44.92^{\circ}C$, incubation time of 10.24 hr, and hexane content of 1.16 mL, whereas that of 1,2-DAG (26.78 area%) was maximized at $56.32^{\circ}C$, 6.95 hr, and 1.04 mL, respectively. Predicted maximum total DAG content was 45.09 area% at $53.82^{\circ}C$, 8.03 hr, and 1.08 mL, while production conditions of MAG (9.57 arae%) were $64.14^{\circ}C$, 7.00 hr, and 0.13 mL. At variables of $54.07^{\circ}C$, 7.98 hr, and 1.02 mL, maximum content of total DAG+MAG predicted by RSM was 53.54 area%.