• Title/Summary/Keyword: and biotin

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An Automated Fiber-optic Biosensor Based Binding Inhibition Assay for the Detection of Listeria Monocytogenes

  • Kim, Gi-Young;Morgan, Mark;Ess, Daniel;Hahm, Byoung-Kwon;Kothapalli, Aparna;Bhunia, Arun
    • Food Science and Biotechnology
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    • v.16 no.3
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    • pp.337-342
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    • 2007
  • Conventional methods for pathogen detection and identification are labor-intensive and take days to complete. Biosensors have shown great potential for the rapid detection of foodborne pathogens. Fiber-optic biosensors have been used to rapidly detect pathogens because they can be very sensitive and are simple to operate. However, many fiber-optic biosensors rely on manual sensor handling and the sandwich assay, which require more effort and are less sensitive. To increase the simplicity of operation and detection sensitivity, a binding inhibition assay method for detecting Listeria monocytogenes in food samples was developed using an automated, fiber-optic-based immunosensor: RAPTOR (Research International, Monroe, WA, USA). For the assay, fiber-optic biosensors were developed by the immobilization of Listeria antibodies on polystyrene fiber waveguides through a biotin-avidin reaction. Developed fiber-optic biosensors were incorporated into the RAPTOR to evaluate the detection of L. monocytogenes in frankfurter samples. The binding inhibition method combined with RAPTOR was sensitive enough to detect L. monocytogenes ($5.4{\times}10^7\;CFU/mL$) in a frankfurter sample.

Novel Vectors for the Convenient Cloning and Expression of In Vivo Biotinylated Proteins in Escherichia coli

  • Cho, Eun-Wie;Park, Jung-Hyun;Na, Shin-Young;Kim, Kil-Lyong
    • BMB Reports
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    • v.32 no.5
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    • pp.497-501
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    • 1999
  • Biotinylation of recombinant proteins is a powerful tool for the detection and analysis of proteins of interest in a large variety of assay systems. The recent development of in vivo biotinylation techniques in E. coli has opened new possibilities for the production of site-specifically biotinylated proteins without the need for further manipulation after the isolation of the recombinantly expressed proteins. In the present study, a novel vector set was generated which allows the convenient cloning and expression of proteins of interest fused with an N-terminal in vivo biotinylated thioredoxin (TRX) protein. These vectors were derived from the previously reported pBIOTRX vector into which was incorporated part of the pBluescript II+phagemid multiple cloning site (MCS), amplified by PCR using a pair of sophisticated oligonucleotide primers. The functionality of these novel vectors was examined in this system by recombinant expression of rat transforming growth factor-$\beta$. Western-blot analysis using TRX-specific antibodies or peroxidase-conjugated streptavidin confirmed the successful induction of the fusion protein and the in vivo conjugation of biotin molecules, respectively. The convenience of molecular subcloning provided by the MCS and the effective in vivo biotinylation of proteins of interest makes this novel vector set an interesting alternative for the production of biotinylated proteins.

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Expression of Cyclooxygenase-2 (COX-2) in Colorectal Adenocarcinoma: an Immunohistochemical and Histopathological Study

  • Mahmoud, Abla Sayed;Umair, Ayesha;Azzeghaiby, Saleh Nasser;Alqahtani, Fahad Hussain;Hanouneh, Salah;Tarakji, Bassel
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.16
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    • pp.6787-6790
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    • 2014
  • Background: The aim of this study was to evaluate cyclooxygenase-2 (COX-2) immunoreactivity in colorectal adenocarcinomas and to find correlations with different pathological features. Materials and Methods: This study included 35 cases of colorectal carcinoma foir which surgical colectomy specimens were collected. Immunohistochemical staining of COX-2 (cyclooxygenase-2) is done by using the Streptavidin-biotin technique. Results: This work reveals that COX-2 is positive in most cases of colorectal carcinoma and negative in normal colon tissue with statistically non significant relations between COX-2 immunostaining and different pathological features. Conclusions: Our data suggest over expression of COX-2 protein in colorectal carcinoma in contrast to normal mucosa, with a possible role in cell proliferation in carcinogenesis.

Immunoreactive ${\beta}$-Endorphin in Female Reproductive Organs (여성 생삭기에 있어서의 ${\beta}$-Endorphin에 관한 면역조직학적 연구)

  • Kim, Jung-Gu;Min, Eung-Gi;Moon, Shin-Yong;Lee, Jin-Yong;Chang, Yoon-Seok
    • Clinical and Experimental Reproductive Medicine
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    • v.15 no.1
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    • pp.53-60
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    • 1988
  • The aim of this study was to examine the presence of ${\beta}$-endorphin in female reproductive organs. A total of 104 fresh tissue samples were obtained from normal ovary, tube, endometrium, placenta, amniotic membrane and umbilical cord, and immunostained by the method using biotin-streptoavidin amplified system. The results were as follows: 1. In reproductive age, corpus luteum only showed ${\beta}$-endorphin immunostained cells but no cells in ovaries during proliferative phase of menstrual cycle were stained. 2. Secretory endometrium revealed positive reactions in the cytoplasm of glandular epithelial cells and around the vessels, while proliferative endometrium negative reactions. 3. All the tissues of menopausal women were negative to ${\beta}$-endorphin antibody. 4. In the pregnant women, there are no ${\beta}$-endorphin containing cells in the placenta, amniotic membrane and umbilical cord regardless of gestational age.

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Evaluation of the Menus of Senior Welfare Centers for Home-bound Elderly (재가 노인들을 위한 노인복지관의 급식 식단 평가)

  • Jung, Eun-kyung;Park, Soojin;Kim, Jin Sook
    • Journal of the Korean Dietetic Association
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    • v.23 no.4
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    • pp.363-373
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    • 2017
  • This study examined the characteristics of the foodservice menu items offered at senior welfare centers to provide information on Korean senior menu development. A total of 514 lunch menu items were collected from 27 senior welfare centers in April, July, October and January. The most frequently served staple foods, soups, and side dishes were multi-grain rice, seaweed soup, Bulgogi, Kimchi, and liquid yogurt. The proportions of carbohydrate, protein, and lipids of total energy serving of senior welfare centers were 59.8%: 16.7%: and 22.8%, respectively. The nutrients served at less than 40% of the Recommended Nutrient Intake (RNI) and Adequate Intake (AI) of Dietary Reference Intakes for Koreans (KDRIs) were chloride (1.0%), vitamin D (1.3%), biotin (1.7%), magnessium (4.5%), Iodine (7.5%), pantothenic acid (8.0%), vitamin E (12.5%), vitamin $B_6$ (20.0~21.4%), vitamin K (21.1~24.3%), and water (35.7~39.7%). The nutrients served in excess of the daily intake goal and RNI were iron (98.9~127.1%), sodium (104.9%), and copper (1,100.0%).

Detection of Protein Molecules by Electrical Current Response Using Two-Electrode Method

  • Lyu, Hong-Kun;Woo, Sung-Ho;Han, Yoon-Soo;Lee, Hee-Ho;Shin, Jang-Kyoo
    • Journal of Sensor Science and Technology
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    • v.20 no.2
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    • pp.90-95
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    • 2011
  • In order to protect human lives from disease, various biosensors having the potential to analyze a variety of biomolecules have been utilized. Biosensors constitute one of the most promising ways to monitor and detect various biomolecules corresponding to diseases. In this study, we demonstrate that the reaction of streptavidin molecules with biotin on a gold electrode can be detected using the twoelectrode method with a gold electrode and a platinum reference electrode. We also show the characteristics of the electrical current response. While detecting 2-${\mu}M$ streptavidin molecules dissolved in phosphate buffered saline(PBS) solution, we found that an analytical biosensor can operate on the principle of detecting an antigen-antibody reaction event of protein molecules using the two-electrode method. We think that the "potential step" method might be useful to detect the occurrence of any antigen-antibody reactions and can be combined with other devices or ICs such as BJTs, MOSFETs, and OP-amps for the detection of biomolecules of diseases.

Study on Bead-based Microbiochip and Analytical System for Protein Detection

  • Kim, Min-Soo;Chung, Woo-Jae;Cho, Su-Hyung;Park, Sung-Soo;Kim, Byung-Gee;Lee, Young-Sik;Kim, Yong-Kweon
    • Proceedings of the KIEE Conference
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    • 2002.11a
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    • pp.60-63
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    • 2002
  • This paper presents bead-based microbiocihps to detect and separate target proteins. Micro beads coated with capture proteins were introduced into a microchamber, and target proteins flowing across the chamber were bound and concentrated. The chip was connected with an external fluid system. Bead surfaces were double-coated with photo-cleavable linkers and capture proteins. The proteins bound on the beads were photo-separated under UV irradiation, and excited to be measured in fluorescence. $38{\sim}50{\mu}m$ sized polystyrene beads were used. SOGs(silicon-on-glass) were used to fabricate the microchip having glasses bonded on both sides. 100 ${\mu}m$ thick silicon channel was formed through silicon deep RIE process. The upper glass cover had holed through to have inlets and outlets fabricated by powder-blastings. In this study, biotin and streptavidin were used as capture proteins and detection proteins, respectively. The protein mixtures of streptavidin, HSA(human serum albumin) and ovalbumin were applied for selective detection test.

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Studies on the Production of L-Glutamic Acid by Brevibacterium ammoniagenes (Brevibacterium ammoniagenes에 의한 글루탐산 제조에 관한 연구)

  • Yoo, Young-Jin;Kim, Taik-Yung
    • Korean Journal of Food Science and Technology
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    • v.9 no.1
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    • pp.1-9
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    • 1977
  • A bacterium strain (K-173-10) which was isolated from waste soil of Korea brewing factory, could be grown on acetate as the sole carbon source and accumulated a considerable amount of L-glutamic acid in the medium. This strain was identified as the new species Brevibacterium ammoniagenes. This study was concerned not only with the culture condition for the production of L-glutamic acid and the cell growth, but also with the effects on concentration of various kind of organic substances, growth factors and penicillin. The results obtained were summarized as follow; 1. It was found that the concentrations of acetate and ammonium ions affected the growth of the bacterium as well as its L-glutamate accumulation. The optimum conditions of the composition of grown media for the growth of the bacterium and its glutamic acid production was found to be 40 g/l of total acetate, $100\;{\mu}g/l$ thiamine, $0.5\;{\mu}g/l$ biotin and $1{\sim}2g/l$ corn steep liquor as the growth factors. 2. Organic acid such as succinic acid, malic acid and ${\alpha}-ketoglutaric$ acid inhibited the cell growth as well as its L-glutamic acid production. 3. The penicillin (20 units/ml) stimulated the production of glutamic acid at appropriate incubation period. 4. It was found that this strain could grow in the presence of urea and ammonium acetate but not in other nitrogen sources.

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Production of Laccase by Trametes sp. CJ-105 (Trametes sp. CJ-105에 의한 Laccase 생산)

  • 오광근;김현수;이재흥;전영중
    • Microbiology and Biotechnology Letters
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    • v.27 no.6
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    • pp.477-483
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    • 1999
  • For Trametes sp. CJ-105, a kind of white-rot fungi which was collected from the mountain of Korea and was proven to be effective in decolorizing a wide range of structurally different synthetic dyes, the optimum conditions for mycelial growth and laccase(E.C. 1.10.3.2) production were investigated. Among various carbon sources, glucose showed the highest potential for the mycelial growth and laccase production, the optimum concentration being 2% glucose. For the nitrogen source, asparagine was good for the mycelial growth, while ammonium tartrate for laccase production(optimum concentration: 0.04%). The addition of thiamine and biotin increased both th emycelial growth and laccase production. When 2,5-xylidine was added as an inducer after the first day of culture, the production of alccase was seven-times higher than that in the absence of the inducer. The optimum pH and temperature conditions for laccase production by Trametes sp. CJ-105 were pH 5.0 and $25^{\circ}C$, respectively. In the 5L fermentation, the production of laccase reached a maximum of 340U/ml at the time when the ammonium ion was being rapidly depleted.

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Rapid Detection of Salmonella enteritidis in Pork Samples with Impedimetric Biosensor: Effect of Electrode Spacing on Sensitivity

  • Kim, Gi-Young;Moon, Ji-Hea;Hahm, Bung-Kwon;Morgan, Mark;Bhunia, Arun;Om, Ae-Son
    • Food Science and Biotechnology
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    • v.18 no.1
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    • pp.89-94
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    • 2009
  • Frequent outbreaks of foodborne illness have been increasing the awareness of food safety. Conventional methods for pathogen detection and identification are labor-intensive and take days to complete. Some immunological, rapid assays are developed, but these assays still require prolonged enrichment steps. Recently developed biosensors have shown potential for the rapid detection of foodborne pathogens. In this study, an impedimetric biosensor was developed for rapid detection of Salmonella entritidis in food sample. To develop the biosensor, an interdigitated microelectrode (IME) was fabricated by using a semiconductor fabrication process. Anti-Salmonella antibodies were immobilized based on neutravidin-biotin binding on the surface of the IME to form an active sensing layer. To evaluate the effect of electrode gap on sensitivity of the sensor, 3 types of sensors with different electrode gap sizes (2, 5, and $10{\mu}m$) were fabricated and tested. The impedimetric biosensor could detect $10^3\;CFU/mL$ of Salmonella in pork meat extract with an incubation time of 5 min. This method may provide a simple, rapid, and sensitive method to detect foodborne pathogens.