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A Gene-Tagging System for Monitoring of Xanthomonas Species

  • Song, Wan-Yeon;Steven W. Hutcheson;Efs;Norman W. Schaad
    • The Plant Pathology Journal
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    • v.15 no.3
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    • pp.137-143
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    • 1999
  • A novel chromosomal gene tagging technique using a specific fragment of the fatty acid desaturase-like open reading frame (des-like ORF) from the tox-argK gene cluster of Pseudomonas syringae pv. phaseolicola was developed to identify Xanthomonas spp.released into the environment as biocontrol agents. X. campestris pv. convolvuli FB-635, a pathogen of Convolvulus arvensis L., (bindweed), was chosen as the organism in which to develop and test the system. A 0.52 kb DES fragment amplified from P. syringae pv. phaseolicola C-199 was inserted into pGX15, a cosmid clone containing a 10.3 kb Eco RI-HindIII fragment derived from the xanthomonadin biosynthetic gene cluster contained in plasmid pIG102, to create a pigG::DES insertion. The 10.8 kb EcoRI-BamHI fragment carrying the pigG:: DES insertion was cloned into pLAFR3 to generate pLXP22. pLXP22 was then conjugated into X. campestris pv. convolvuli FB-635 and the pigG::DES insertion integrated into the bacterial chromosome by marker exchange. Rifampicin resistant, tetracycline sensitive, starch hydrolyzing, white colonies were used to differentiate the marked strain from yellow pigmented wild-type ones. PCR primers specific for the unique DES fragment were used for direct detection of the marked strain. Result showed the marked strain could be detected at very low levels even in the presence of high levels of other closely related or competitive bacteria. This PCR-based DES-tagging system provides a rapid and specific tool for directly monitoring the dispersal and persistence of Xanthomonas spp.released into the environment.

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Clinical features of Senior-Loken syndrome with IQCB1/NPHP5 mutation in a Filipino man

  • Chiu, Harold Henrison C.;Sucaldito, Ma. Sergia Fatima P.;Maceda, Ebner Bon G.;Montemayor, Jan Andre S.;Tamondong-Lachica, Diana R.
    • Journal of Genetic Medicine
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    • v.17 no.1
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    • pp.39-42
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    • 2020
  • The Senior-Loken syndrome was first described in 1961 as an oculo-renal disease consisting of familial juvenile nephronophthisis and Leber congenital amaurosis. It is a rare autosomal recessive disorder with a prevalence of 1:1,000,000 caused by mutations in nine genes (NPHP 1-8 and NPHP 10). Ocular manifestations (e.g., photophobia, nystagmus, and extreme hyperopia) occur within the first few years of life while renal manifestations (e.g., formation of multiple cysts impairing kidney function and end-stage renal disease) appear in late childhood to adolescence. Here, we report a case of a Filipino male presenting with rotatory nystagmus and progressive deterioration of vision since childhood. He had congenital amaurosis and juvenile nephronophthisis that progressed to end stage renal disease by age 19. All laboratory and imaging findings were consistent with chronic kidney disease. Molecular genetic testing of ciliopathy-related genes was performed revealing a homozygous mutation in exon 11 of the IQCB1/NPHP5 gene, c.1090C>T (p.Arg364). This sequence change created a premature translational stop signal resulting in a truncated protein product, nephrocystin-5 and its consequent loss of function. His symptoms eventually improved with initiation dialysis. The prognosis of Senior-Loken syndrome remains dismal and a high index of suspicion, early diagnosis and timely intervention of renal complications are warranted.

Optimal Conditions of Reaction Flavor for Synthesis of Crab-like Flavorant from Snow Crab Cooker Effluent (홍게 자숙액으로부터 게향 제조를 위한 반응향의 최적화)

  • Ahn, Jun-Suck;Jeong, Eun-Jeong;Cho, Woo-Jin;Cha, Yong-Jun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.1
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    • pp.128-134
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    • 2014
  • To develop a crab-like flavorant from snow crab cooker effluent (SCCE, $20^{\circ}Brix$), optimal reaction conditions were determined using response surface methodology (RSM) combined with reaction flavoring technology (RFT). Using five variables (proline, glycine, arginine, methionine, fructose), RSM based on a five-level central composite design was applied to evaluate sensory acceptance (odor, taste, and overall acceptance) as dependent variables. A model equation obtained from RSM showed 0.88 of R-square for odor, 0.90 for taste, and 0.95 for overall acceptance with 0.07 lack of fit in overall acceptance (P<0.05). Odor score (predicted value) was 7.21 in the saddle point. Optimal flavoring conditions for making a crab-like flavorant were as follows: addition of 0.29 g of proline, 0.63 g of glycine, 0.61 g of arginine, 0.02 g of methionine, and 1.07 g% (w/v) of fructose into SCCE with RFT (90 min at $130^{\circ}C$). Odor score obtained under optimal conditions was 7.56, which was higher than the predicted value.

Quality Characteristics of Chun Ma (Gastrodiae rhizoma) Beverage Prepared Using Concentrated Extracts (천마추출액 농축 비율을 달리하여 제조한 천마음료의 품질특성)

  • Lee, Su-Won;Moon, Hye-Kyung;Moon, Jae-Nam;Yoon, Won-Jung;Kim, Gwi-Young
    • Food Science and Preservation
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    • v.17 no.1
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    • pp.58-65
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    • 2010
  • We investigated the effect of Chun ma (Gastrodiae rhizoma) concentrated extract on the quality of mixed beverages. Chun ma beverages prepared using different concentrated extracts were divided into four groups: GCE 5 ($5^{\circ}$ Brix concentrated extracts) GCE 10 ($10^{\circ}$ Brix concentrated extracts) GCE 15 ($15^{\circ}$ Brix concentrated extracts) and GCE 20 ($20^{\circ}$ Brix concentrated extracts). The pH values ranged from a low of 4.37 in GCE 5 to a high of 4.68 in GCE 20. Soluble solid levels in GCE 20 ($19.6^{\circ}$ Brix) were higher than in the other samples. The b (yellowness) scores and the total phenolic contents of all samples increased with increasing extract concentration. The highest total phenolic contents were seen in GCE 20 samples at 232.23 mg%. Samples did not differ markedly in antiradical activity (75.07-76.00% DPPH inhibition). Free sugar levels in GCE 20 samples and organic acid concentrations of GCE 15 samples were higher than those of other preparations. Free amino acid and mineral contents of all samples increased with increasing extract concentration. The levels of free amino acids were in the order Glu > Gly > Ser > Arg > Hylys, and the Glu content was 249.15 ug/100 g for GCE-20 samples and 61 ug/100 g in GCE-5 products. The mineral contents of all samples were in the order K > Na > Mg > Ca. Higher scores for color, flavor, and overall acceptability were found in GCE 5 products compared with other extracts. These results indicate that Chun ma beverage can be prepared in various ways, as commercially desired, with reference to the above characteristics of Chun ma materials.

Ethanol Production an Immobilized Themotolerant Mutant of Brettanomyces custersii H1-39 from Wood Hydrolyzate Media (목질계 당화액배지로부터 고온내성 변이주 Brettanomyces custersii H1-39의 고정화에 의한 에탄올생산)

  • 박승원;홍영기;김승욱;홍석인
    • Microbiology and Biotechnology Letters
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    • v.28 no.3
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    • pp.172-179
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    • 2000
  • Bretlanomyces C!tstersii Hl-39 mutant was immobilized with various caniers. Immobilized mutant Hl-39 produced more ethanol and showed higher productivity and cell concentration than those of free 81-39 in 3.4% hydrolyzate of wood-chips at different temperatures ($37^{\circ}C$, $40^{\circ}C$ and $43^{\circ}C$). At $37^{\circ}C$, ethanol concentration produced by mutant H1-39 immobilized in Ca-alginate and ARG(l % Ca-alginate, 1.67% bentonite, 0.33% glutaraldehyde) bead were higher than those produced by the other earners (ACG ; 1 % CaHalginate. ] .67% celite R-634 , 0.33% glutaraldehyde, ABP ; 1 % Ca-alginate. 1.67% bentonite, 0.33% pectin. ACP: 1 % Ca-alginate, ] .67% celiLe R-634, 0.33% pecLin). The highest value of productivity(l.23 ) was obtained by using ABG beads. At $40^{\circ}C$, ethanol conccntration and productivity obtained by ABC beads ,>,"ere 15.2 glL and 0.84 gl L.h, respectively, which showed the highest value compared to other carriers. Particularly, productivity of ilmnobilized ceIl was increased up to 90% as compared to that offree cell. On the other hand, ABP(l % Ca-alginate+L67% bentonile+O.33% pectin) beads gave the best resulLs at $43^{\circ}C$ for production of ethanol and productivity, which were 13.8 g!l and 0.77 g/l h, respectively.ively.

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Chemical Composition and Antimicrobial Activities of Houttuynia cordata Thunb. (어성초의 화학성분 및 항미생물 활성)

  • Kim, Keun-Young;Chung, Dong-Ok;Chung, Hee-Jong
    • Korean Journal of Food Science and Technology
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    • v.29 no.3
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    • pp.400-406
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    • 1997
  • Chemical composition and antimicrobial activity of Houttuynia cordata Thunb. were investigated to develop a natural food preservative from it. Aspartic acid, glutamic acid, glycine and arginine were major amino acids of Houttuynia cordata Thunb., but were present in a trace amount. Free sugars were composed of glucose, fructose, sucrose and maltose and major fatty acids were linolenic acid, linoleic acid, oleic acid, and palmitic acid. Mineral elements were potassium, calcium, magnesium, iron, zinc, and copper. Antimicrobial activities were shown in acidic, neutral and phenolic fraction of Houttuynia cordata Thunb., but not in basic fraction. Among the four fractions, neutral fraction showed the strongest antimicrobial activities against microorganisms tested, such as Pseudomnas aeruginosa, Micrococcus luteus, Bacillus subtilis, Aspergillus parasiticus and Aspergillus flavus. Minimum inhibitory concentration (MIC) and minimum lethal concentration (MLC) of the neutral fraction varied according to microoganisms tested. The lowest values of MIC (0.0075 g eq./mL) and MLC (0.10 g eq./mL) were obtained from Pseudomonas aeruginosa.

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The First Korean case of combined oxidative phosphorylation deficiency-17 diagnosed by clinical and molecular investigation

  • Kim, Young A;Kim, Yoo-Mi;Lee, Yun-Jin;Cheon, Chong Kun
    • Clinical and Experimental Pediatrics
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    • v.60 no.12
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    • pp.408-412
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    • 2017
  • Combined oxidative phosphorylation deficiency-17 (COXPD-17) is very rare and is caused by homozygous or compound heterozygous mutations in the ELAC2 gene on chromosome 17p12. The ELAC2 gene functions as a mitochondrial tRNA processing gene, and only 4 different pathogenic mutations have been reported in ELAC2-associated mitochondrial dysfunction involving oxidative phosphorylation. Affected patients show various clinical symptoms and prognosis, depending on the genotype. We report a novel mutation in the ELAC2 gene (c.95C>G [p.Pro32Arg], het), in an infant with COXPD-17 who presented with encephalopathy including central apnea and intractable epilepsy, and growth and developmental retardation. During hospitalization, consistently elevated serum lactic acid levels were noted, indicative of mitochondrial dysfunction. The patient suddenly died of shock of unknown cause at 5 months of age. This is the first case report of COXPD-17 in Korea and was diagnosed based on clinical characteristics and genetic analysis.

Inhibitory Effects of Various Edible Plants and Flavonoids from the Leaves of Cedrela sinensis on Human Immunodeficiency Virus Type 1 Protease

  • Park, Jong-Cheol;Hur, Jong-Moon;Park, Ju-Gwon;Kim, Hyun-Joo;Kang, Kyeong-Hee;Park, Myeong-Rak;Song, Sang-Ho
    • Preventive Nutrition and Food Science
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    • v.5 no.3
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    • pp.170-173
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    • 2000
  • The inhibitory effect of extracts from 15 edible plants on the protease of human immunodeficiency virus (HIV) type 1 was investigated. Protease activity was determined by incubating the extracts in a reaction mixture containing protease and substrate His-Lys-Ala-Arg-Val-Leu-(p-NO$_2$-Phe)-Glu-Ala-Nle-Ser-NH$_2$ to inhibit proteolytic cleavage. Of various plants tested, the leaves of Cedrela sinensis inhibited the HIV-1 protease by 42% at a concentration of 100$\mu\textrm{g}$/ml. A major flavonoid isolated from the leaves of C. sinensis, quercetin 3-O-$\alpha$-L-rhamnoside showed inhibitory activity of 19% at a concentration of 100$\mu$M.

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Partial characterization of a 17 kDa protein of Clonorchis sinensis

  • Chung, Young-Bae;Chung, Byung-Suk;Choi, Min-Ho;Chai, Jong-Yil;Hong, Sung-Tae
    • Parasites, Hosts and Diseases
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    • v.38 no.2
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    • pp.95-97
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    • 2000
  • A 17 kDa protein from Clonorchis sinensis adults was purified by a procedure including Sephacryl S-200 HR gel filtration and Q-Sepharose anion exchange chromatography. The protein was proved to be a cysteine protease as it showed hydrolytic activity toward Cbz-Phe-Arg-AMC in the presence of dithiothreitol and was inhibited by specific inhibitors such as iodoacetic acid or trans epoxy-succinly-L-leucyl-amido(4 guanidino) butane. The polyclonal antibody raised against the protein reacted to 17 kDa proteins of trematodes such as Paragonimuf westermani, Fasciola hepatica, Opisthorchis viverrini, Gymnophalloides seoi, and Metagonimus yokogawai. The antibody recognized the 17 kDa and 16 kDa cysteine proteases purified from C. sinensis, P. westemani, and G. seoi as well. These results suggest that the 17 kDa protein may be a cysteine protease commonly present in trematodes.

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Citrus unshiu Water Extract Inhibits Trypsin-induced $TNF-{\alpha}$ and Tryptase Productions by Blocking the ERK Phosphorylation and Trypsin Activity

  • Kang, Ok-Hwa;Kim, Dae-Ki;Lee, Young-Mi
    • Natural Product Sciences
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    • v.10 no.5
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    • pp.211-216
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    • 2004
  • Citrus unshiu (Rutaceae) has long been known as an anti-inflammatory and anti-allergic agent. In the present study, the inhibitory effect of CUWE (Citus unshiu water extract) on the production of $TNF-{\alpha}$ and tryptase was examined. In addition, a possible mechanism for the inhibition of trypsin-stimulated human leukemic mast cell-1 (HMC- 1 ) activation was determined. To do so, $TNF-{\alpha}$ production from the HMC-1 cells that were stimulated by trypsin (100 nM) in the presence or absence of CUWE $(10,\;100,\;and\;100\;{\mu}g/ml)$ was measured by enzyme-linked immunosorbent assay (ELISA) and reverse transcription-PCR. The tryptase production was evaluated by reverse transcription-PCR. Extracellular signal-regulated kinase (ERK) activation was analyzed by Western blot. Trypsin activity was measured by using Bz-DL-Arg-p-nitroanilide (BAPNA) as substrate. Results showed that the CUWE inhibited production of both $TNF-{\alpha}$ and tryptase from the trypsin-stimulated HMC-1 in a dose-dependent manner. The CUWE a1so inhibited the ERK phosphorylation and trysin activity. These results indicate that the CUWE had an inhibitory effect on $TNF-{\alpha}$ and the tryptase productions by blocking the ERK phosphorylation and trypsin activity.