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Development and Characterization of Expression Vectors for Corynebacterium glutamicum

  • Lee, Jinho
    • Journal of Microbiology and Biotechnology
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    • v.24 no.1
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    • pp.70-79
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    • 2014
  • In an attempt to develop a variety of expression vector systems for Corynebacterium glutamicum, six types of promoters, including $P_{tac}$, $P_{sod}$, $P_{sod}$ with a conserved Shine-Dalgarno (SD) sequence from C. glutamicum, $P_{ilvC}$, $P_{ilvC}$ with a conserved SD-1 ($P_{ilvC-M1}$), and $P_{ilvC}$ with a conserved SD-2 ($P_{ilvC-M2}$), were cloned into a modified shuttle vector, pCXM48. According to analysis of promoter strength by quantitative reverse transcription PCR, $P_{sod}$ and $P_{sod-M}$ were superior to tac and ilvC promoters in terms of transcription activity in C. glutamicum. All of the promoters have promoter activities in Escherichia coli, and $P_{sod-M}$ displayed the highest level of transcriptional activity. The protein expression in constructed vectors was evaluated by measuring the fluorescence of green fluorescent protein (GFP) and SDS-PAGE. C. glutamicum harboring plasmids showed GFP fluorescence with an order of activity of $P_{ilvC}$ > $P_{ilvC-M1}$ > $P_{sod}$ > $P_{ilvC-M2}$ > $P_{sod-M}$, whereas all plasmids except pCSP30 with $P_{sod}$ displayed fluorescence activities in E. coli. Of them, the strongest level of GFP was observed in E. coli with $P_{sod-M}$, and this seems to be due to the introduction of the conserved SD sequence in the translational initiation region. These results demonstrate that the expression vectors work well in both C. glutamicum and E. coli for the expression of target proteins. In addition, the vector systems harboring various promoters with different strengths, conserved SD sequences, and multiple cloning sites will provide a comfortable method for cloning and gene expression, and consequently contribute to the metabolic engineering of C. glutamicum.

Physicochemical properties of sausage manufactured with carp (Carassius carassius) muscle and pork

  • Lee, Sol-Hee;Joe, Sung-Duck;Kim, Gye-Woong;Kim, Hack-Youn
    • Journal of Animal Science and Technology
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    • v.62 no.6
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    • pp.903-911
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    • 2020
  • The purpose of this study was to compare the physicochemical properties of sausage manufactured with carp (Carassius carassius) muscle and pork. Sausages were prepared using either 100% pork sausage (P10), a mixture of 50% pork and 50% carp muscle (P5C5), or 100% carp muscle (C10). The quality of the sausage emulsion was determined by analyzing approximate composition, pH, instrumental color, cooking yield, water holding capacity (WHC), viscosity, and texture profile analysis (TPA). Moisture content of cooked C10 was significantly higher than that of P10 or P5C5 (p < 0.05); however, protein content of cooked P10 was significantly higher than that of C10 (p < 0.05). The pH of uncooked and cooked C10 was significantly higher than that of P10 and P5C5 (p < 0.05). The cooking yield, WHC, and texture profile analysis of C10 were higher than those of P10 and P5C5 (p < 0.05). In addition, the viscosity of uncooked C10 was higher than that of P10 and P5C5. These results suggest that carp muscle can enhance sausage quality with respect to pH, WHC, cooking yield, viscosity, and TPA.

C-P-N-D Ecological System and ICCT (Information, Communication, Contents Technology) (C-P-N-D 생태계와 ICCT (Information, Communication, Contents Technology))

  • Choi, ChangHyeon
    • Journal of Digital Convergence
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    • v.12 no.3
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    • pp.7-16
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    • 2014
  • This study aims to analyze the key logic of the current C-P-N-D ICT ecological system, to find out the shortcomings of the current system, and then to offer policy suggestions for the establishment of a new creative contents industry ecological system; that is, ICCT (Information, Communication, Contents and Technology) System.

Viscogram Pattern of Korean Rice Flours (우리나라 쌀의 점도 특성)

  • Kim, Sung-Kon;Kim, Sang-Soon
    • Applied Biological Chemistry
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    • v.28 no.3
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    • pp.142-148
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    • 1985
  • Pasting properties of 35 traditional (Japonica) and 23 high-yielding (J/Indica) rice varieties having various hydration rates at $23^{\circ}C$ were investigated. Amylograms showed that high-yielding rice varieties had higher values for peak (P), hot-paste (H) and cold-paste (C) viscosities than those for traditional rice varieties. P had a highly negative correlation with H/P, C/P and C/H. P of traditional rice varieties was positively and negatively correlated with protein amylose contents, respectively. However, H and C showed no correlation with protein or amylose content. No correlation was observed between amylograph indices and hydration rate of rice grain.

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Arylation of Styrene by Palladium Acetate-Phosphine Complexes

  • 황박영애;황성원
    • Bulletin of the Korean Chemical Society
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    • v.18 no.2
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    • pp.218-221
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    • 1997
  • When phenylation of styrene was carried out in the presence of Pd(OAc)2 and PPh3 in benzene, trans-stilbene was obtained in good yield (566%) with high selectivity (98%) under mild condition (55 ℃, 50 psi O2, 20 h). Since trans-stilbene could be produced not only from benzene but also from phenyl group of PPh3 by migration of its phenyl group to Pd, the competitiveness of benzene and the migratory aptitude of aryl group of triarylphosphine toward styrene has been investigated with various phosphines (PR3: P(p-C6H4CH3)3, P(p-C6H4OCH3)3, P(p-C6H4F)3, P(p-C6H4Cl)3, P(C6H5)3, P(C6H11)3, P(OC4H9n)3, P(CH2C6H5)3 and P(C6F5)3). The yield and selectivity toward trans-stilbene are increased as the basicity of the phosphines increases. The composition of arylated olefin from arylphosphine, in turn, increases as the electronegativity of the substituent on the aryl group of arylphosphines increases.

Characterization of Ovarian Cytochrome $P450_{C17}$ (17 ${\alpha}-hydroxylase$/17,20-lyase) in Rana dybowski (북방산 개구리 난소의 Cytochrome $P450_{C17}$ 유전자 특성)

  • Kang, Hae-Mook
    • Development and Reproduction
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    • v.10 no.2
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    • pp.127-133
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    • 2006
  • [ $17\;{\alpha}-hydroxylase/17,20-lyase(P450_{C17})$ ] is the key enzyme mediating the conversion of progesterone to $17\;{\alpha}-hydroxyprogesterone$, ultimately to androstenedione during steroidogenesis. R. dybowskii's ovarian $P450_{C17}$ cDNA was cloned to understand the regulatory mechanism of ovarian steroidogenic pathway at the molecular level in amphibian. A 2.5kb cDNA clone encoding a single open-reading frame with a 519 deduced amino acid was isolated with the screening of ovarian cDNA library. This sequence contained the three highly conserved domains as seen in $P450_{C17}$ of other species. The comparison of amino acid sequence of Rana $P450_{C17}$ with other animal's $P450_{C17}$ showed relatively high identity with 76% in Xenopus, 63% in chicken, 60% in rainbow trout, and 45% in human. Phylogenic analysis also indicated that Rana $P450_{C17}$ gene was evolutionary well conserved among vertebrate. Northern analysis indicated that the two different sizes of $P450_{C17}$ transcripts with approximately 2.5 and 3.6kb were detected in ovary tissue, but not in other tissues. The expression vector of Rana $P450_{C17}$ clearly showed the $17\;{\alpha}-hydroxylase$ activity converting the exogenous progesterone into $17\;{\alpha}-hydroxyprogesterone$ in the nonsteroidogenic COS-1 cells. Therefore, Rana $P450_{C17}$ cDNA is very useful to investigate the molecular mechanism of the ovarian steroidogenesis in amphibian.

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Assessment of Biochemical Profiles in Premenopausal and Postmenopausal Women with Breast Cancer

  • Yadav, Naval Kishor;Poudel, Bibek;Thanpari, C.;Koner, Bidhan Chandra
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.7
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    • pp.3385-3388
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    • 2012
  • Objective: The study was conducted to assess biochemical profiles in premenopausal and postmenopausal women having breast cancer. Materials and Methods: A hospital based case control study was carried out at Manipal Teaching Hospital (MTH), Pokhara, Nepal. The analysed variables were age, metabolic profile including total cholesterol, triglycerides, HDL-C, LDL-C, blood sugar, insulin concentration, C-peptide, HbA1c and selenium. Descriptive statistics and testing of hypothesis were used for the analysis using EPI INFO and SPSS 16 software. Results: In premenopausal women, significant differences were noted for total cholesterol (P value <0.001), triglycerides (P value 0.002), HbA1c level (P value <0.001), insulin concentration (P value 0.030), C-peptide concentration (P value 0.001), and selenium (P value <0.001) between cases and controls. Insignificant results were found for HDL-C (P value 0.749), LDL-C (P value 0.933), blood sugar (P value 0.59) and BMI (P value 0.746). Similarly, significant difference in total cholesterol (P value <0.001), triglycerides (P value 0.001), LDL-C (P value <0.001), HDL-C (P value 0.025), blood sugar (P value <0.001), insulin concentration (P value <0.001), c-peptide concentration (P value <0.001), HbA1c level (P value <0.001) and selenium (P value <0.001) were observed for postmenopausal patients and controls. Conclusions: Assessing metabolic changes and their management may be important for control of breast cancer and increased survival.

Substrate chain-length specificities of polyhydroxyalkanoate synthases PhaC1 and PhaC2 from Pseudomonas aeruginosa P-5 (Pseudomonas aeruginosa P-5에 존재하는 polyhydroxyalkanoate synthase PhaC1과 PhaC2의 기질특이성)

  • Woo, Sang Hee;Lee, Sun Hee;Rhee, Young Ha
    • Korean Journal of Microbiology
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    • v.52 no.4
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    • pp.455-462
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    • 2016
  • Pseudomonas aeruginosa P-5 is an unusual organism capable of synthesizing polyhydroxyalkanoates (PHAs) consisting of 3-hydroxyvalerate (3HV) and medium-chain-length (MCL) 3-hydroxyalkanoate (3HA) monomer units when C-odd alkanoic acids are fed as the sole carbon source. Evaluation of the substrate chain-length specificity of two P. aeruginosa P-5 PHA synthases ($PhaC1_{P-5}$ and $PhaC2_{P-5}$) by heterologous expression of $PhaC1_{P-5}$ and $PhaC2_{P-5}$ genes in Pseudomonas putida GPp104 revealed that $PhaC2_{P-5}$ incorporates both 3HV and MCL 3HAs into PHA, whereas $PhaC1_{P-5}$ favors only MCL 3HAs for polymerization. In order to obtain $PhaC2_{P-5}$ mutants with altered substrate specificity, site-specific mutagenesis for $PhaC2_{P-5}$ was conducted. Amino acid substitutions of $PhaC2_{P-5}$ at two positions (Ser326Thr and Gln482Lys) were very effective for synthesizing copolymers with a higher 3HV fraction. When recombinant P. putida GPp104 harboring double mutated $phaC2_{P-5}$ gene ($phaC2_{P-5}QKST$) was grown on nonanoic acid, 2.5-fold increase of copolymer content with 3.8-fold increase of 3HV fraction was observed. The $phaC2_{P-5}QKST$-containing Ralstonia eutropha PHB-4 supplemented with valeric acid also produced copolymers consisting of 3HV and 3-hydroxyheptanoate with a high 3HV fraction. These results suggest that recombinants containing $phaC2_{P-5}QKST$ could be useful for production of new PHA copolymers with improved material properties.

RELATING GALOIS POINTS TO WEAK GALOIS WEIERSTRASS POINTS THROUGH DOUBLE COVERINGS OF CURVES

  • Komeda, Jiryo;Takahashi, Takeshi
    • Journal of the Korean Mathematical Society
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    • v.54 no.1
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    • pp.69-86
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    • 2017
  • The point $P{\in}{\mathbb{P}}^2$ is referred to as a Galois point for a nonsingular plane algebraic curve C if the projection ${\pi}_P:C{\rightarrow}{\mathbb{P}}^1$ from P is a Galois covering. In contrast, the point $P^{\prime}{\in}C^{\prime}$ is referred to as a weak Galois Weierstrass point of a nonsingular algebraic curve C' if P' is a Weierstrass point of C' and a total ramification point of some Galois covering $f:C^{\prime}{\rightarrow}{\mathbb{P}}^1$. In this paper, we discuss the following phenomena. For a nonsingular plane curve C with a Galois point P and a double covering ${\varphi}:C{\rightarrow}C^{\prime}$, if there exists a common ramification point of ${\pi}_P$ and ${\varphi}$, then there exists a weak Galois Weierstrass point $P^{\prime}{\in}C^{\prime}$ with its Weierstrass semigroup such that H(P') = or , which is a semigroup generated by two positive integers r and 2r + 1 or 2r - 1, such that P' is a branch point of ${\varphi}$. Conversely, for a weak Galois Weierstrass point $P^{\prime}{\in}C^{\prime}$ with H(P') = or , there exists a nonsingular plane curve C with a Galois point P and a double covering ${\varphi}:C{\rightarrow}C^{\prime}$ such that P' is a branch point of ${\varphi}$.

Effect of Incubation Temperature and pH on Chlamydospores Germination of Cylindrocarpon destructans Causing Root Rot of Panax ginseng (인삼 뿌리썩음병균 Cylindrocarpon destructans의 후막포자 발아에 미치는 배양온도 및 pH의 효과)

  • 조대휘;유연현
    • Journal of Ginseng Research
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    • v.25 no.3
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    • pp.136-140
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    • 2001
  • Effects of incubation temperature and pH on chlamydospore germination of Cylindrocarpon destrcutans (isolate CY-9802) causing root rot of Panax ginseng were studied. Germination rate of the chlamydospores on Czapek solution agar(CSA) was higher than on potato dextrose agar(PDA) at the incubation temperatures tested. The chlamydospores were able to be germinated at range of 5$\^{C}$ to 30$\^{C}$ after 48 hours incubation on CSA. Germination rate was 53.2∼6.27% at range of 15$\^{C}$ to 25$\^{C}$, and the optimum temperature was 20$\^{C}$, whereas they were very low at 30$\^{C}$ on PDA. Germination rate was 43.6% to 47.9% at range of 10$\^{C}$ to 20$\^{C}$, and the optimum temperature was 20$\^{C}$ as well. They were able to be germinated at pH of 5.2 to 8.1 on CSA and 5.2 to 7.2 on PDA. Optimum pHs for the germination on CSA and PDA were from 6.4 to 8.2 and from 5.2 to 6.0, respectively. Mycelial color of the fungus on CSA was pale brown at pH from 5.2 to 6.0 and white from pH 6.4 to 8.1, while it was typical dark brown ar range of pH 5.2 to 7.1 and brown at pH 7.2 on PDA after 21 days incubation.

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