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Isolation and Identification of Weissella kimchii from Green Onion by Cell Protein Pattern Analysis

  • Kim, Tae-Woon;Lee, Ji-Yeon;Song, Hee-Sung;Park, Jong-Hyun;Ji, Geun-Eog;Kim, Hae-Yeong
    • Journal of Microbiology and Biotechnology
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    • v.14 no.1
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    • pp.105-109
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    • 2004
  • This study was conducted to investigate the potential origin of Weissella species, which were found in ingredients of kimchi, such as salted Chinese cabbage, radish, green onion, red pepper powder, pickled shrimps, garlic, and ginger. Ten strains of Weissella species (Weissella thailandensis, W. kimchii, W. koreensis, W. minor, W. halotolerans, W. hellenica, W. kandleri, W. confusa, W. viridescens, and W. paramesenteroides) and lactic acid bacteria isolated from ingredients of kimchi were analyzed by SDS-PAGE of whole-cell proteins. Several strains with patterns identical to those of Weissella kimchii were isolated from green onion. On the basis of biochemical characteristics and 16S rDNA sequence comparisons, these strains were identified as Weissella kimchii, suggesting green onion as a major origin of Weissella kimchii found in kimchi.

Studies on the Characteristics of Anti-Zona Antibody III. Examination of Biochemical Characteristics of Porcine Zone Pellucidae and Anti-Zona Antibody (항투명대 항체의 특성에 관한 연구 III. 돼지투명대와 항투명대 항체의 생화학적 특성 검토)

  • 김은영;박세필;신경순;정길생;김종배
    • Korean Journal of Animal Reproduction
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    • v.14 no.2
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    • pp.125-131
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    • 1990
  • These experiments were carried out to investigate of the molecular characteristics of porcine zona pellucidae and to examine the reactivity of anti-zona antibody by SDS-PAGE, Immunoblotting and Immunoprecipitation. The results obtained in these experiments were summarized as follows : 1. The result obtained by SDS-PAGE of porcine zona pellucidae indicated that it composed of several units with molecular weight ranging 55,000-110,000. 2. In order to see the reactivity of antibodies to zona pellucidas, immunoblotting was applied. The results indicated that polyclonal antibodies to porcine and mouse zona reacted with porcine zona. While monoclonal antibody to porcine did not react with the procine zona enough to show a clear band on a gel. 3. Labelling of porcine zonae with 125I was performed using the Iodogen method, the radioactivity and the percent incorporation of 125I into porcine zonae were approximately 26,000 cpm/10${mu}ell$ and 16, respectively. 4. Measurements of radioactivity and O.D value for Immunoprecipitates produced by reaction of 125I-porcine zona with anti-zona antisera were confirmed that existence of reactivity of monoclonal antibody to porcine zona although its reactivity was lower than that of polyclonal antibodies, and reconfirmed that cross-reactivity of polyclonal antibody of mouse zona with porcine zona.

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Effect of concentration, pressure, and cut-off size on removing phospholipase A2 in bee venom by ultrafiltration (Ultrafilteration을 이용한 봉독의 phospholipase A2 제거에서 농도, 압력, 분자크기의 영향)

  • Lee, Ji-Yeon;Kim, Il Kwang;Lee, Jong Soo;Kim, Eui Kyung;Kim, Choul-Goo
    • Analytical Science and Technology
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    • v.27 no.6
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    • pp.277-283
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    • 2014
  • The effects of concentration, pressure, and molecular sige on removing allergenic substance (phospholipase $A_2$) in bee venom by ultrafiltration were investigated. The membrane pore sizes were selected based on the molecular weight of the main compounds. The conditions of concentration and pressure were selected randomly. As results, we obtained the optimum condition (1 mg/mL, 20 psi, 10,000 dalton) for removing $PLA_2$ at constant concentration of melittin and apamin and confirmed the separation results by HPLC and SDS-PAGE.

Identification of the Gene Products Responsible for F Plasmid Partitioning

  • Kim, Sung-Uk;Kazuo Nagai;Gakuzo Tamura;Yu, Ju-Hyun;Bok, Song-Hae
    • Journal of Microbiology and Biotechnology
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    • v.3 no.4
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    • pp.256-260
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    • 1993
  • DNA subfragments, sopA, sopB and sopC which help to maintain the stability of an ori C plasmid, were derived from a mini-F plasmid DNA (EcoRI restriction fragment f5) after digestion with restriction endonuclease, and cloned in the vector plasmid pBR322. The recombinant plasmids obtained were introduced into E. coli KY7231 and E. coli CSR603 strains, and proteins specified by the mini-F fragments were analysed by SDS-PAGE. Two proteins encoded by the F fragments were detected, and their molecular weights were 41,000 and 37,000 daltons. Fluorography after one and two dimensional gel electrophoresis of the lysates showed that these two proteins had been overproduced in the cells which were allowed to incorporate radioactive amino acid after plasmid amplification by chloramphenicol treatment. The isoelectric points of sopA and sopB proteins were 6.6 and 7.0, respectively.

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Purification and Biophysicochemical Properties of the Virus Causing Necrotic Spot Symptom on Korean Cactus (한국산 접목 선인장에서 분리한 괴사 반점 원인 바이러스의 생물리화학적 특성)

  • 이철호;나용준;한정헌
    • Korean Journal Plant Pathology
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    • v.13 no.2
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    • pp.73-78
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    • 1997
  • A virus was purified from Korean cactus (Chamaeaereus silverstrii f. Variegata Hort) showing necrotic spots and its biophysicochemical properties were examined. The typical necrotic spots were observed on healthy looking cactus inoculated with the purified virus and the various symptoms from necrotic spots to systemic were observed on 17 species of test plats used, which suggests that the purified virus has the normal infectivity and wide host range. Ultraviolet absorption scanning analysis revealed that the viral preparation had the typical curve with shoulder at 290 nm and its ratio of absorbances at 260 nm to 280 nm and extinction coefficient was 1.156 and 2.86, respectively. The virus represented one distinct and homogeneous band and molecular weight of viral coat protein was determined to 17, 000 Dalton by SDS-PAGE. The virus revealed rod-shaped virion (325$\times$20 nm) by electron microscopy. Based on these results, it has been thought that the virus purified in this study is the pathogenic agents causing necrotic spot symptom on Korean cacuts and one of the member of tobamovirus family.

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Studies on the development of the tick (Haemaphysalis longicornis)-vaccine (I) - Immune responses on the crude soluble - (진드기 백신 개발을 위한 기초연구(I) - 수용성 항원에 대한 면역반응에 관하여 -)

  • Jeong, Woo-seog;Kang, Seung-won;Choi, Eun-jin;Yoon, Yong-dhuk
    • Korean Journal of Veterinary Research
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    • v.36 no.3
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    • pp.693-698
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    • 1996
  • Haemaphysalis longiscornis is the common cattle tick of great economic importance in Korea. Chemical control using dips or sprays has been the traditional method of attempting to kill these ticks during the infestation period. However, the presence of resistant forms to chemical, the rising costs of acaricides and environmental problems have made it almost impossible to use these chemicals on a regular basis according to the pest problem. For this reason, vaccination against ticks and breeding for host resistance against ticks are being studied. In order to determine the common proteins and antigens according to developmental stages, SDS-PAGE and western blotting were performed. In SDS-PAGE 103.3kD and 98.3kD proteins were observed as common proteins, and these proteins were observed as common antigens in western blotting. Unimmunized rabbits were infestated three times with H longicornis. The weight of the second and the third engorged ticks were 0.153g and 0.104g respectively. This weight is 69% and 47% of the first engorged ticks weight respectively. Immunized rabbits by adult ticks antigen and control were infested with H longicornis. The control taked 3-4 days to fully engorge, but the immunized rabbits taked about 7 days. So adult tick antigen may be effective to render the immunity to host.

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Expression of Bacillus thringiensis HD-1 gene in rhizobacteria Pseudomonas fluorescens KR164 (근권 길항세균 Pseudomonas fluorescens KR164에 Bacillus thuringiensis HD-1 유전자의 삽입과 발현)

  • Kim, Yeong-Yil;Rhee, Young-Hwan;Kang, Heun-Soo
    • Applied Biological Chemistry
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    • v.35 no.4
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    • pp.227-231
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    • 1992
  • The plasmids pSUPBT and pSUPBTR were constructed with a vector pSUP2021 and the BT toxin gene in the plasmid pES 1. The plasmids constructed were introduced into the antagonistic rhizobacteria P. fluorescens KR164 by conjugation and P. fluorescens having pSUPBT and pSUPBTR were named P. fluorescens KR164(pSUPBT)#2, KR164(pSUPBT)#3, KR164(pSUPBTR)#2 and KR164(pSUPBTR)#3, respectively. The BT toxin gene were identified in all transformants by Southern hybridization and the final product of BT toxin gene was identified only in P. fluorescens KR164(pSUPBTR)#3 by SDS-PAGE. This crystal toxin protein were also observed in electron microscopy.

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SOLVING BIOANALYTICAL PROBLEMS BY THE METHOD OF MATRIX-ASSISTED LASER DESORPTION IONIZATION MASS SPECTROMETRY (MALDI-MS)

  • Zhao, Shankai;Zhong, Feng;Zhu, Zhihua
    • Analytical Science and Technology
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    • v.8 no.4
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    • pp.865-868
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    • 1995
  • The method of matrix-assisted laser desorption ionization mass spectrometry has been used to solve some bioanalytical problems, which is difficult to analyse by general methods. For the selection of proper laser wavelength and matrices, eight matriees was used with laser wavelength of 226 and 355nm. The result shows that with wavelength of 355nm better results could be obtained with most of the matrices. The molecular weight of eytochrome C, which was seperated by gel electrophoresis and electro-blotted onto NC membrane is determined by MALDI. The accuracy is better than 0.1%, which is much higher than that of SDS-PAGE. Protein mixture extracted from crude peanut oil is directly determined by MALDI. The molecuiar weight of its three components are determined, and the result also demonstrated that these proteins are in free manner. As proteins arc in 2S bond, with the traditional method, SDS-PAGE, it is not able to decide whether protein exists in combination mode or in free manner. In the technique of two phase aquesous solution, which is used for separating biomaterials, water soluble polymers stained with dyes are used in this technique. By the use of MALDI the number or the dye molecules react with the polymer PEG molecule are determined, and that is difficult to determined by other methods.

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Effect of Ginseng Saponin Fractions on Phagocytosis and Chemotaxis of Murine Macrophages (대식세포의 식세포활동과 화학주성에 대한 인삼분획물의 영향)

  • Shin, Eun-Kyoung;Kim, Sei-Chang
    • The Journal of Natural Sciences
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    • v.8 no.2
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    • pp.27-34
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    • 1996
  • The phagocytosis and chemotaxis of murine macrophages after treated with saponin fractions are investigated. Phagocytic appearance against yeast was photographed by dying with Wright-Giemsa. Phagocytic activity of peritoneal macrophage was invreased in diol saponin treatment by 48% and was decreased in total saponin treatment by 35%. The ingestion of alveolar macrophage was increased by 50% maximally. Peritoneal chemotactic activity was shown in 17% increases and only diol saponin had effect in alveolar macrophage by 16%. According to SDS-PAGE method the contents of actin did not show any alterations.

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Identification of Allergens in Pork Meat (돼지고기 중 알레르기 유발성분의 동정)

  • 정혜주;박재현;김재희;김영옥;정승태;김진호;조은득;조대현;노건웅
    • YAKHAK HOEJI
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    • v.45 no.1
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    • pp.39-45
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    • 2001
  • The pork meat has been reported as one of the food occurring allergic reactions predominantly to korean. To identify the potential food allergens in pork meat, sera were collected from 25 allergic patients to the pork meat and 10 allergic patients not to pork meat as well as 5 normal subjects after skin prick test and open food challenge test. Crude extracts were prepared by blending raw pork meat in phosphate buffered saline (pH 7.0) and the heat treatment on crude extracts was carried to characterize sensibility of the allergens to heat. ELISA was performed to determine specific IgE antibody levels of allergic patients to pork meat, and resulted in twofold higher mean value than that of tolerated patients. Extracted proteins from pork meat was separated with SDS-PAGE followed by immunoblotting using sera from pork sensitive patients and control subjects, respectively. The IgE binding response to pork meat by immunobots correlated with quantitative specific IgE value of each person. Immunoblots showed four prominent IgE-binding bands (66, 60, 50, 44 kDa) in crude extract, but two bands of those (60, 44 kDa) were heat-labile. These results suggest that most prominent allergens from pork meat are four components(66, 60, 50, 44 kDa) in korean and the heat treatment on allergen is additional parameter to characterize allergen.

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