• Title/Summary/Keyword: MALDI-TOF/MS

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Muscle Proteome Analysis for the Effect of Panax Ginseng Extracts in Chicken: Identification of Proteins Using Peptide Mass Fingerprinting

  • Jung, K.C.;Yu, S.L.;Lee, Y.J.;Choi, K.D.;Choi, J.S.;Kim, Y.H.;Jang, B.G.;Kim, S.H.;Hahm, D.H.;Lee, J.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.7
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    • pp.922-926
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    • 2005
  • The present study was aimed to investigate proteome affected by Panax ginseng extracts in chicken muscles. The whole muscle proteins from chicken fed boiled extracts of 0% (control), 1%, 3%, and 5% Panax ginseng in water were separated by two-dimensional electrophoresis (2-DE) gels using immobilized non-linear gradient (pH 3-10) strips. More than 300 protein spots were detected on silver staining gels. Among them, four protein spots were distinctively up-regulated by Panax ginseng treatments and further investigated by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). The obtained MS data were searched against SwissProt database using the Mascot search engine. The up-regulated proteins were finally identified as $\alpha$-tropomyosin (2 spots), triosephosphate isomerase, and one unknown protein. Based on the known functions of the identified proteins, they are highly related to muscle development and enhanced immunity in chickens. These proteins can give valuable information of biochemical roles for Panax ginseng in chicken meats.

Isolation and Characterization of Six Microorganisms from the Digestive Tract of the Cricket Gryllus bimaculatus (쌍별귀뚜라미(Gryllus bimaculatus) 소화기관에서 분리한 6종류의 특성규명)

  • Kwon, Kisang;Lee, Eun Ryeong;Yoo, Bo-Kyung;Ko, Young Hwa;Shin, Hyojung;Choi, Ji-Young;Kwon, O-Yu
    • Journal of Life Science
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    • v.27 no.9
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    • pp.1040-1046
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    • 2017
  • We describe the isolation and characterization of six different intestinal microorganisms from the digestive tract of the cricket Gryllus bimaculatus. Based on 16S rRNA gene sequences, we obtained six isolates belonging to four different genera: Staphylococcus, Bacillus, Citrobacter, and Proteus. All the isolates were resistant to ampicillin. Ampicillin is an irreversible inhibitor of the enzymeetranspeptidase, which is needed to make bacterial cell walls. None of the isolates were resistant to kanamycin, which binds to the 30S subunit of the bacterial ribosome and then inhibits total protein synthesis. Gram staining was conducted, in addition to morphological classification under a microscope. Four grampositive isolates and two gram-negative isolates were detected. The gram-positive isolates were GL1 (round shaped, 2 am in diameter), GL2 (rod shaped, $2.5{\mu}m$ in length), GL3 (rod shaped, $2{\mu}m$ in length), and GL4 (round shaped, $1.5{\mu}m$ in diameter). The gram-negative isolates were GL5 (rod shaped, $2{\mu}m$ in length) and GL6 (rod-shaped, $2.5{\mu}m$ in length). Notably, two of the isolates, GL2 and GL4, secreted specific extracellular proteins. These were determined by MALDI-TOF-MS spectral analysis to be a 87 kDa collagenase, 56 kDa hypothetical protein, and 200 kDa hypothetical protein. The six isolates in this study could be used for various biotechnological applications and pest management, both in the field and in greenhouse systems. In addition, it would be interesting to determine the relationship between these isolates and their host.

Isolation and characterization of the outer membrane vesicle (OMV) protein from Vibrio anguillarum O1 (Vibrio anguillarum O1이 생산하는 Outer Membrane Vesicle (OMV)의 분리 및 OMV 내의 단백질 특성)

  • Hong, Gyeong-Eun;Kim, Dong-Gyun;Min, Mun-Kyeong;Kong, In-Soo
    • Journal of Marine Bioscience and Biotechnology
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    • v.2 no.2
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    • pp.123-125
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    • 2007
  • Vibrio anguillarum is a gram-negative bacterium that causes vibriosis in approximately 80 different fish species. V. anguillarum produces several exotoxins are correlated with the pathogenesis of vibriosis. This study is focused on the composition of the outer membrane vesicle. Most of gram-negative bacteria produce outer membrane vesicle (OMV) during cell growth. OMV was formed from the outer membrane surface of cell and than released to extracellular environment. OMV consists of outer membrane lipids, outer membrane protein (OMP), LPS, and soluble periplasmic components. Also, they contain toxins, adhesions, and immunomodulatory. Many gram-negative bacteria were studied out forming OMV. In Vibrio sp., formation of OMV by electron microscopy has been reported from V. cholerae and V. parahaemolyticus. In present study, we isolated OMV from V. anguillarum and OMV protein was separated by SDS-PAGE. Magor band was sliced and analyzed by MALDI-TOF. The major protein band of 38kDa was identified as OmpU by MALDI-TOF MS analysis.

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Detection of methicillin-resistant S. pseudintermedius (MRSP) isolated from dogs and cats in Kyungpook Area (경북지역 개와 고양이에서 methicillin-resistant Staphylococcus. pseudintermedius (MRSP) 검출률 조사)

  • Byun, Jun Ho;Sohn, Jun Hyung;Lee, Young Mi;Jeong, Hyo Hoon;Oh, Tae Ho
    • Korean Journal of Veterinary Service
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    • v.41 no.4
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    • pp.229-237
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    • 2018
  • Identification of antibiotic resistant bacteria isolated from animals is necessary for seeking a proper treatment and for preventing the spread of the bacteria among animals. Methicillin-resistant Staphylococcus pseudintermedius (MRSP) is of worldwide concern in veterinary medicine. This study was conducted to investigate the antimicrobial susceptibility of Staphylococcus pseudintermedius (S. pseudintermedius) isolated from dogs and cats. Out of the total number of 150 specimens, 35 isolates (23.3%) were identified as S. pseudintermedius when tested by MALDI-TOF MS (Matrix Assisted Laser Desorption Ionization-Time of Flight Mass Spectroscopy). Among them, 5 isolates (14.3%) were the case of MRSP. In the results of the antimicrobial susceptibility test, the isolates of S. pseudintermedius were susceptible to amikacin (100%), cephalothin (100%), vancomycin (100%), amoxicillin (85.7%), oxacillin (85.7%), enrofloxacin (82.8%), chloramphenicol (80%). On the other hand, they were resistant to penicillin (71.4%), tetracycline (48.6%), ampicillin (37.1%), kanamycin (31.5%), erythromycin (31.4%), respectively.

Comparison of Matrices for Optimal Analysis of Synthetic Polymers Using MALDI-TOF Mass Spectrometry

  • Yoo, Hee-Jin;Kim, Duck-Hyun;Choi, Yoon-Ji;Choi, Jung Hoon;Park, Moonhee;Shin, DongJin;Oh, YoonSeok;Kim, YangSun;Cho, Kun
    • Mass Spectrometry Letters
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    • v.11 no.4
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    • pp.77-81
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    • 2020
  • Characterization of the various chemical aspects of composite polymers is important for quality control of manufactured polymers. In this study, we compared three suitable matrices (α cyano-4-hydroxycinnamic acid [CHCA], 2,5 dihydroxy benzoic acid [2,5-DHB], and dithranol), to characterize various synthetic polymers by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Although the spectra obtained with the CHCA and 2,5-DHB matrices were generally good, in certain samples ghost peaks disappeared only when dithranol was used as the matrix. Furthermore, we examined the use of sodium trifluoroacetate (NaTFA) as an additive to reduce interference by metals and copolymers in the spectra. In conclusion, appropriate selection of a matrix, according to the characteristics of the polymer, and the use of additives to improve sensitivity are important considerations for polymer analysis and development.