• Title/Summary/Keyword: Amino-

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Construction of a Fluorescently Labeled Infectious R Peptide-Less Moloney MLV Molecular Clone for Analysis of Syncytium (합포체 분석을 위해 R 펩타이드가 결여된 형광 표지 Moloney 마우스레트로바이러스 Molecular Clone 제조)

  • Lee, Yong-Jin;Park, Jin-Woo;Lee, Kyu-Jun;Bae, Eun-Hye;Park, Sung-Han;Lim, Ji-Hyun;Kim, Sae-Ro-Mi;Jung, Yong-Tae
    • Korean Journal of Microbiology
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    • v.45 no.3
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    • pp.246-250
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    • 2009
  • Retroviruses enter host cells by membrane fusion between the viral Env proteins on the virus membrane and a virus receptor on the cellular membrane. The envelope protein of the ecotropic Moloney murine leukemia virus is synthesized as a gp85 precursor and is proteolytically cleaved into an extracellular surface unit (SU) and the transmembrane protein (TM). The cytoplasmic tail (16 amino acid; R peptide) of the TM protein is further cleaved by the viral protease during virion maturation. Unlike the wild type Env protrin bearing the R peptide, R peptide-truncated Envelope induces syncytia in susceptible cells. To understand the mechanism of R peptidetruncated Env in syncytium formation, R peptide-truncated Env expressing full-length molecular clone containing EGFP in PRR (proline rich region) of Env was constructed. This molecular clone induced syncytia in transfected NIH3T3 cells, fluorescence was detected in the cytoplasm and at the plasma membrane, while the nuclei did not stain and appeared black by fluorescence microscopy. Interestingly, virions with truncated envelope produced from transfected NIH3T3 cells induced syncytia in NIH3T3 cells, but fluorescence was not detected in the same infected cells. It is believed that cell-free viruses direct the fusion of neighboring cells without infection. Our data suggests that use of EGFP-tagged envelope for monitoring syncytium is a sensitive and convenient method. We also found that virion incorporated the R peptide-truncated Env is able to induce the formation of syncytia by fusion from without.

Changes in Quality of Citron Juice by Storage and Extraction Conditions (유자과즙의 저장 및 착즙조건에 따른 품질변화)

  • Park, Kee-Jai;Jung, Sung-Won;Kim, Jong-Hoon;Jeong, Jin-Woong
    • Applied Biological Chemistry
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    • v.38 no.2
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    • pp.141-146
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    • 1995
  • Changes of physicochemical properties of citron juice prepared by two different extraction methods, rotary-crushing and belt-pressing method, were investigated during the storage at $5^{\circ}C$ and $-20^{\circ}C$. Temperature drop of citron juice extracted by belt-pressing method was faster than that of citron juice prepared by rotary-crushing method and its freezing point was $0.8{\sim}0.9^{\circ}C$. During the storage, pH of stored citron juice with rotary-crushing method was increased up to 3.5 after 6 months storage while that of citron juice extracted by belt-pressing method was not changed significantly during the same storage time. Acidity of rotary-crushed citron juice was reduced a little more than that of belt-pressed citron juice during the storage. However, changes of soluble solid content were influenced largely by the storage temperature than by the extraction method. Contents of formol nitrogen and vitamin C were reduced remarkably in all of stored citron juice and $92{\sim}82%$ of farmol nitrogen and $72{\sim}43%$ of vitamin C were remained after 6 months of storage. Among the changes of color value, L values were reduced in the whole stored citron juice and a and b value had a different change pattern respectively according to the extraction and storage temperature. Changes in the content of both amino acid and fatty acid compositions was also observed after same storage period. Especially, in the case of change of fatty acid composition, content of linoleic acid and linolenic acid were reduced after 6 months storage, while those of palmitic acid, stearic acid and oleic acid were increased.

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Clinical Findings and Gene Analysis of 3-Methylcrotonyl-CoA Carboxylase Deficiency (3-methylcrotonyl-CoA carboxylase 결핍증의 임상 양상과 유전자 분석)

  • Lee, Seung Eun;Ahn, Hee Jae;Lee, Jeongho;Lee, Dong Hwan
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.15 no.1
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    • pp.1-8
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    • 2015
  • Purpose: 3-methylcrotonyl CoA carboxylase deficiency (3MCCD) is leucine metabolic disorder caused by mutation in MCCC1 or MCCC2 gene. Clinical manifestations are variable, ranging from fatal neonatal onset to asymptomatic individuals. There is no retrospective study of Korean patients undergoing long-term treatment for 3MCCD. We reported this study to find out clinical symptoms and gene analysis of 3MCCD patients. Methods: This study was based on data of patients diagnosed with 3MCCD in Soonchunhyang university hospital between April 2009 and September 2013. We report clinical, enzymatic and mutation data of 3MCCD patients found by newborn screening. Results: In tandem mass spectrometry, 3-OH-isovalerylcarnitine (C5OH) of all patients increased. And all 7 patients were elevated 3-methylcrotonylglycine (3MCG) and 3-hydroxyisovaleric acid (3HIVA) in urine. MCCC mutation was identified in 2 patients and MCCC2 was mutated in 5 patients. We found mutation occurred in 8 different parts of nucleotide and such mutation caused 7 different types of changes in amino acid. All patients are on medication of L-carnitine and L-glycine. 4 patients are taking biotin. And 4 patients are eating leucine free formula. After starting treatment, there were no significant changes of urine 3MCG and 3HIVA levels. Conclusions: According to our data, MCCC2 gene mutation was more common than MCCC1 gene mutation. But the level of 3HIVA or 3MCG in urine has no correlation with phenotype. All patients has no symptoms and are shown normal development.

Genes of Wild Rice (Oryza grandiglumis) Induced by Wounding and Yeast Extract (상처와 효모추출물 처리조건에서 유발되는 야생벼 유전자 스크린)

  • Shin, Sang-Hyun;Im, Hyun-Hee;Lee, Jai-Heon;Kim, Doh-Hoon;Chung, Won-Bok;Kang, Kyung-Ho;Cho, Sung-Ki;Shin, Jeong-Sheop;Chung, Young-Soo
    • Journal of Life Science
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    • v.14 no.4
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    • pp.650-656
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    • 2004
  • Oryza grandiglumis (CCDD, 2n=48), one of the wild rice species, has been known to possess fungal-,bacterial-, and insect-resistance against sheath blight, rice blast, bacterial leaf blight and brown plant hopper (Nilaparvata lugens). To rapidly isolate differentially expressed genes responding to fungal and wounding stress, wounding and yeast extract were treated to O. grandiglumis for 24 hrs. Suppression subtractive hybridization (SSH) method was used to obtain differentially expressed genes from yeast extract and wounding treated plants. Seven hundreds and seventy six clones were obtained by subcloning PCR product, and colony array and screening were carried out using radio-isotope labeled cDNA probes prepared from the wounding and yeast extract treated plants. One hundred and fifteen colonies were confirmed as true positive ones. Average insert size of the clones were ranged from 400 bp to 700 bp and all the inserts were sequenced. To decide the identity of those clones, sequences were analyzed by sequence homology via GenBank database. The homology search result showed that 68 clones were matched to the genes with known function; 16 were related to primary metabolism, 5 to plant retrotransposons, 5 to defense related metallothionein-like genes. In addition to that, others were matched to various genes with known function in amino acid synthesis and processing, membrane transport, and signal transduction, so on. In northern blot analysis, induced expressions of ogwfi-161, ogwfi-646, ogwfi-663, and ogwfi-695 by wounding and yeast extract treatments were confirmed. The result indicates that SSH method is very efficient for rapid screening of differentially expressed genes.

Effects of Brewery Meal-Based Fermented Feedstuff Supplemented with Aspergillus Oryzae or Saccharomyces Cerevisiae on Ruminal Microorganism (Aspergilus Oyzae 혹은 Saccharomyces Cerevisiae를 첨가하여 제조한 맥주박 위주 발효사료가 반추위 미생물에 미치는 영향)

  • Park, Byung-Ki;Ra, Chang-Six;Kim, Byong-Wan;Oh, Jin-Seok;Shin, Jong-Suh
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.25 no.4
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    • pp.307-318
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    • 2005
  • This study was conducted to examine effects of brewery meal-based fermented feedstuff supplemented with Aspergillus oryzae(AO) or Saccharomyces cerevisiae(SC) on luminal micro-organism of Korean native cattle. Two cows equipped with luminal cannulas were used as experimental animals. Experiment was done with three treatment groups: $71.5\%$ of commercial feed and $28.5\%$ of com silage(control): $45.0\%$ of commercial feed, $26.5\%$ of fermented feedstuff supplemented with AO and $28.5\%$ of corn silage(TAO): $45.0\%$ of commercial feed, $26.5\%$ of fermented ffedstuff supplemented with SC and $28.5\%$ of corn silage(TSC). The number of total viable bacteria (p<0.05), anaerobic fungi and protozoa(p<0.05) was higher in TAO and TSC than in control. The number of proteolytic bacteria(p<0.05), cellulolytic bacteria and xylan fermenters tended to be higher in TAO and TSC than in control. The dry matter recovery (DMR) of protozoa was higher in TAO and TSC than in control(p<0.05). The crude protein (CP) content of total microbes and protozoa was higher in TSC than in control and TAO (p<0.05). The CP content of bacteria was higher in TAO and TSC than in control(p<0.05). The ether extract(EE) content of the total microbes was higher in TAO than in control and TSC(p<0.05), and the EE of protozoa and bacteria were higher in TSC than in control and TAO(p<0.05). The ratio of essential amino acids of total microbe was higher in control than in TAO and TSC(p<0.05). The ratio of methionine and alanine of bacteria was higher in TAO and TSC than in control(p<0.05). The results suggested that the feeding of fermented feedstuff supplemented with AO or SC had an influence on the numbers of ruminal microorganism and the changes of microbial body composition.

Quality Characteristics of Rice $Makgeolli$ Prepared by Mashing Types (담금유형에 따른 쌀 막걸리 술덧의 품질특성)

  • Park, Chan-Woo;Jang, Se-Young;Park, Eun-Ji;Yeo, Soo-Hwan;Jeong, Yong-Jin
    • Korean Journal of Food Science and Technology
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    • v.44 no.2
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    • pp.207-215
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    • 2012
  • Six different mashing types ((A) $koji$+purified enzyme, (B) $koji$+crude enzyme, (C) $koji$+$nuruk$, (D) $koji$+purified enzyme+$nuruk$, (E) $koji$+crude enzyme+$nuruk$, (F) purified enzyme+$nuruk$) had been established, according to fermentation agents and a mixing rate of rice $makgeolli$, in this study. The alcohol content was the highest in the mashing type (C), which was 13.6%, followed by (D) 13.5%, (A) 13.1%, (B) 12.9%, (E) 12.7% and (F) 12.1%. The reducing sugar content of (A) was the highest with 401.6 mg% and those of (B), (C), (D) and (F) were between 337.3- 380.9 mg%. The alcohol components were found and tended to increase during the fermentation. The oligo-saccharides content was the highest in (D) with 1251.3 mg%, which was followed by (E) 1,219.2 mg%, (C) 1,141.4 mg%, (A) 1,049.9 mg% and 973.8 mg% in (B). The total free amino acid was highest in (B) with 781.4 mg% and followed by (C) 703.2 mg%, (D) 702.6 mg%, (E) 678.7 mg%, (A) 630.4 mg% and (F) 328.7 mg% in order. There were 16 different types of volatile flavor components, in the mashing types (A) and (B), in addition to 15 different types of those in type (C), as well as 14 different types of those in (D), (E) and (F). There were significant differences in the overall preference between the type (A) and (C).

Physiological Activity and Nutritional Composition of Pleurotus Species (느타리속 버섯류의 영양성분 및 생리활성)

  • Um, Su-Na;Jin, Gyoung-Ean;Park, Kye-Won;Yu, Young-Bok;Park, Ki-Moon
    • Korean Journal of Food Science and Technology
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    • v.42 no.1
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    • pp.90-96
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    • 2010
  • In this study, the anti-oxidant, anti-tumorigenic, anti-hypertensive, anti-thrombic, anti-diabetic, and anti-inflammatory properties of 18 different species of genus Pleurotus were investigated. In addition, the amino acid, $\beta$-glucan, and polyphenol content were also measured. All species contained more than 20 mg% of polyphenol with the highest contents found in Pleurotus cornucopiae var. citrinopileatus (yellow pleurotus) ($39.13{\pm}0.82\;mg%$). The $\beta$-glucan contents was also the highest in yellow Pleurotus ($37.67{\pm}0.22%$) followed by Won-Hyeong1 (C, $28.75{\pm}0.61%$) and Jang-an PK (A, $27.95{\pm}0.33%$). The yellow Pleurotus exhibited the highest antioxidant activity as assessed by the DPPH scavenging rate with an $IC_{50}$ value of $2.94{\pm}0.44\;mg/mL$. Ethanol extracts from the yellow Pleurotus treated at 1% concentration showed cytotoxic activity up to 36.9% in the human embryonic kidney 293T cell lines. The yellow Pleurotus also showed the highest inhibitory effects on ACE activity ($60.52{\pm}0.2%$). Finally, the yellow Pleurotus exhibited anti-diabetic and anti-inflammatory properties as shown by inhibition of $\alpha$-amyloglucosidase activity ($50.5{\pm}0.8%$) and nitric oxide production ($68.4{\pm}0.3%$). Taken together, our data indicate the yellow pleurotus is a promising functional food ingredients.

Characterization and Cloning of the Gene Encoding Autoregulator Receptor Protein from Streptomyces longwoodensis (Streptomyces longwoodensis로부터 Autoregulator Receptor Protein 유전자의 클로닝 및 특성)

  • Yeo Soo-Hwan;Lee Sung-Bong;Kim Hyun-Soo
    • Microbiology and Biotechnology Letters
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    • v.33 no.2
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    • pp.96-105
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    • 2005
  • For screening of autoregulator receptor gene from Streptomyces longwoodensis, PCR was performed with primers of receptor gene designed on the basis of amino acid sequences of autoregulator receptor proteins with known function. PCR products were subcloned into the BamHIsite of pUC19 and transformed into the E. coli $DH5{\alpha}$. The isolated plasmid from transformant contained the fragment of 100 bp, which was detected on $2\%$ gel after BamHI treatment. The insert, 100 bp PCR product, was confirmed as the expected internal segment of gene encoding autoregulator receptor protein by sequencing. Southern and colony hybridizations with the 100 bp fragment as a probe allowed to select a genomic clone of S. longwoodensis, pSLT harboring a 4.4 kb SphI fragment. Nucleotide sequencing analyses revealed a 651 bp open reading frame(ORF) were isolated protein showing moderate homology ($35{\sim}46\%$) with the ${\Gamma}$-butyrolactone autoregulator receptors from Streptomyces sp., and this ORF was named sltR The sltR/pET-17b plasmid was constructed to overexpress the recombinant SltR protein (rSltR) in E. coli BL21 (DE3)/pLysS, and the rSltR protein was purified to homogeneity by DEAE-Sephacel column chromatography, and DEAE-5PW chromatography (HPLC). The molecular mass of the purified rSltR protein was 55 kDa by HPLC gel-filtration chromatography and 28 kDa by SDS-PAGE, indicating that the rSltR protein is present as a dimer. A binding assay with tritium-labeled autoregulators revealed that the rSltR has clear binding activity with a A-factor type autoregulator as the most effective ligand.

Trend and Further Research of Rice Quality Evaluation (쌀의 품질평가 현황과 금후 연구방향)

  • Son, Jong-Rok;Kim, Jae-Hyun;Lee, Jung-Il;Youn, Young-Hwan;Kim, Jae-Kyu;Hwang, Hung-Goo;Moon, Hun-Pal
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.47
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    • pp.33-54
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    • 2002
  • Rice quality is much dependent on the pre-and post harvest management. There are many parameters which influence rice or cooked rice qualitys such as cultivars, climate, soil, harvest time, drying, milling, storage, safety, nutritive value, taste, marketing, eating, cooking conditions, and each nations' food culture. Thus, vice evaluation might not be carried out by only some parameters. Physicochemical evaluation of rice deals with amy-lose content, gelatinizing property, and its relation with taste. The amylose content of good vice in Korea is defined at 17 to 20%. Other parameters considered are as follows; ratio of protein body-1 per total protein amount in relation to taste, and oleic/linoleic acid ratio in relation to storage safety. The rice higher Mg/K ratio is considered as high quality. The optimum value is over 1.5 to 1.6. It was reported that the contents of oligosaccharide, glutamic acid or its derivatives and its proportionalities have high corelation with the taste of rice. Major aromatic compounds in rice have been known as hexanal, acetone, pentanal, butanal, octanal, and heptanal. Recently, it was found that muco-polysaccharides are solubilized during cooking. Cooked rice surface is coated by the muco-polysaccharide. The muco-polysaccharide aye contributing to the consistency and collecting free amino acids and vitamins. Thus, these parameters might be regarded as important items for quality and taste evaluation of rice. Ingredients of rice related with the taste are not confined to the total rice grain. In the internal kernel, starch is main component but nitrogen and mineral compounds are localized at the external kernel. The ingredients related with taste are contained in 91 to 86% part of the outside kernel. For safety that is considered an important evaluation item of rice quality, each residual tolerance limit for agricultural chemicals must be adopted in our country. During drying, rice quality can decline by the reasons of high drying temperature, overdrying, and rapid drying. These result in cracked grain or decolored kernel. Intrinsic enzymes react partially during the rice storage. Because of these enzymes, starch, lipid, or protein can be slowly degraded, resulting in the decline of appearance quality, occurrence of aging aroma, and increased hardness of cooked rice. Milling conditions concerned with quality are paddy quality, milling method, and milling machines. To produce high quality rice, head rice must contain over three fourths of the normal rice kernels, and broken, damaged, colored, and immature kernels must be eliminated. In addition to milling equipment, color sorter and length grader must be installed for the production of such rice. Head rice was examined using the 45 brand rices circulating in Korea, Japan, America, Australia, and China. It was found that the head rice rate of brand rice in our country was approximately 57.4% and 80-86% in foreign countries. In order to develop a rice quality evaluation system, evaluation of technics must be further developed : more detailed measure of qualities, search for taste-related components, creation and grade classification of quality evaluation factors at each management stage of treatment after harvest, evaluation of rice as food material as well as for rice cooking, and method development for simple evaluation and establishment of equation for palatability. On policy concerns, the following must be conducted : development of price discrimination in conformity to rice cultivar and grade under the basis of quality evaluation method, fixation of head rice branding, and introduction of low temperature circulation.

Effects of Herbal Product (Miracle®) on the Performance, Nutrient Digestibility, Fecal Microflora and Immunoresponse in Weanling Pigs (생약제제(미라클®)첨가가 이유 자돈의 성장률, 영양소 이용율, 분내 미생물균총 및 면역기능에 미치는 영향)

  • Suk, J.C.;Lim , H.S.;Paik, I.K.
    • Journal of Animal Science and Technology
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    • v.45 no.5
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    • pp.767-776
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    • 2003
  • An experiment was conducted to investigate the effects of dietary herbal product(Miracle$^{\circledR}$) supplementation on the performance, nutrient digestibility, fecal microflora and blood parameters in pigs. Forty eight cross bred(Y${\times}$L${\times}$D) weanling pigs (average initial body weight 7.84kg${\pm}$0.17, 28 d old) were used in 35 d feeding trial. Pigs were allotted to 16 raised floor cages(W 35.5cm${\times}$L45cm${\times}$H 55cm). Each treatment had four replications of 3 pigs each. Treatments were: control (T1), CTC 100ppm (T2), Miracle$^{\circledR}$ 0.15% (T3) and CTC 100ppm + Miracle$^{\circledR}$ 0.15% (T4). The herbal product supplementation(T3) tended to improve the weight gain, feed intake, feed/gain but there were no significant differences among treatments. Nutrient digestibility was significantly(P〈0.05) affected by dietary treatments. The pigs fed T3 diet showed the lowest digestibility of DM, crude protein, NFE, P, and amino acids among treatments. Serum cholesterol level of pigs fed T3 diet was lowest and significantly(P〈0.05) lower than those fed antibiotic supplemented diet(T2). Triglyceride and HDL level of the pigs fed T3 diet were also lower than those of others but the differences were not significant. The level of serum IgG was significantly(P〈0.05) different among treatments. The level of serum IgG was highest in T4, followed by T1, T2 and T3. Serum AST(Aspartate aminotransferase) level of the control(T1) was lower than other treatments. Levels of serum total protein, albumin and ALT(Alanine aminotransferase) were not significantly different among treatments. The colony forming units (CFU) of Lactobacillus, Cl. perfringens and E.coli in feces were not significantly different among treatments but those of Cl. perfringens of T2 (CTC 100ppm) showed the lowest count at both 3rd and 5th wk. It is concluded that herbal product Miracle$^{\circledR}$ does not significantly affect growth performance of weanling pigs but it influenced the digestibility, serum IgG, cholesterol and AST level. The combination of the herbal product Miracle$^{\circledR}$ with antibiotic(CTC) showed no synergistic effects.