• Plant Biotechnology Reports
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• 제2권4호
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• pp.233-238
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• 2008

Kadainou R-1, an interspecific hybrid grape derived from red (Vitis ficifolia var. ganebu) and white (V. vinifera cv. Muscat of Alexandria) grapes, accumulates high concentrations of anthocyanin in the berry skin. Hence, the expression of uridine 50 -diphosphate (UDP)-glucose:flavonoid 3-O-glucosyltransferase (UFGT), the key enzyme of the anthocyanin pathway, was examined in the berry skin of Kadainou R-1. As information on gene sequences of V. ficifolia var. ganebu and other wild grape species was unavailable, we performed GeneChip hybridization using biotin-labeled genomic deoxyribonucleic acid (DNA) to investigate how the genomic sequences of V. vinifera varieties and that of V. ficifolia var. ganebu differ. The study showed a lower correlation coefficient between V. vinifera cultivars and V. ficifolia var. ganebu than that among V. vinifera cultivars. The sequences of the UFGT gene derived from both parents of the red and white cultivars were sequenced in Kadainou R1 and revealed that both were expressed irrespective of the fact that it was not expressed in the white grape (male parent).

• 센서학회지
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• 제20권1호
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• pp.35-39
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• 2011

A disposable electrochemical immunosensor system has been developed for the detection of herbicide in aqueous samples. Disposable screen printed carbon electrodes(SPCE) were used as basic electrodes and an enzyme, horseradish peroxidase (HRP), and anti-herbicide antibodies was immobilised on to the working electrode of SPCE by using avidin-biotin coupling reactions. An herbicide-glucose oxidase conjugates have been used for the competitive immunoreaction with sample herbicides. The enzymatic reaction between the conjugated glucose oxidase and glucose added generates hydrogen peroxide, which was reduced by the peroxidase immobilised. The latter process caused an electrical current change, due to direct re-reduction of peroxidase by a direct electron transfer mechanism, which was measured to determine the herbicides in the sample. The optimal operational condition was found to be: $20\;{\mu}gl-1$ deglycosylated avidin loading to the working electrode and working potential +50 mV vs. Ag/AgCl. The total assay time was 15 min after sample addition. The detection limits for herbicides, atrazine and simazine, were found to be 3 ppb and 10 ppb, respectively.

• 한국수산과학회지
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• 제30권6호
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• pp.948-955
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• 1997

Immunomagnetic separation of virus coupled with .reverse transcription-polymerase chain reaction (IMS-PCR) was performed with infectious hematopoietic necrosis virus (IHNV). A DNA fragment of expected size was synthesized in the RT-PCR with total RNA extracted from IHNV inoculated CHSE-214. In a SDS-PAGE analysis, a protein band of over 70kDa was detected from non-infected cells and cells inoculated with IHNV and infectious pancreatic necrosis virus (IPNV). This protein was detected in the Western blot analysis probably because of non-specific reaction to monoclonal antibody against IHNV nucleocapsid protein. In the immunomagnetic separation, magnetic beads coated with monoclonal antibody against the IHNV nucleocapsid protein was incubated with supernatant from IHNV inoculated CHSE-214 cells. During this process, the non-specifically reacting protein could be removed by washing the magnetic bead with PBS in the presence of an external magnetic field, and viral proteins were detected from the remaining, cleaned magnetic beads. It was necessary to extract viral RNA from the captured virus particles before RT-PCR, and no DNA product was detected when the captured virus was only heated 5 min at $95^{\circ}C$. A PCR-product of expected size was synthesized from IMS-PCR with magnetic beads double coated either by goat anti-mouse IgG antibody -monoclonal antibody or streptavidin - biotin conjugated monoclonal antibody.

• 한국미생물·생명공학회지
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• 제5권1호
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• pp.5-12
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• 1977

As a basic study for the application of the spore-tearing lactic acid bacteria to foods, the effects of the sporulating conditions on the growth and sporogenesis were studied were observed. The results obtained are as follow. 1. All carbohydrates added to sporulation media except dextrin decreased the sporulation rate and the thermal resistance of spores. Dextrin stimulated the growth, however, there in no effect on the thermal resistance. 2. As nitrogen source, the protein hydrolysates such as peptone, casamino acid were effective to obtain were spores of the increased thermal resistance. 3. Ca$\^$＋＋/, Mn$\^$＋＋/ of the metal ions added to casamino acid containing medium validly increased the total growth, sporulation rate and thermal resistance. Its optimum concentration was 40 ppm each. 4. Biotin of vitamines had an effect on the total growth, sporulation and thermal resistance of spores. Its optimum concentration was 30${\gamma}$/ml. 5. The resistant spores required the adequate maturation period, more than 36 hours, sufficient aeration. and optimum temperature, 37∼45$^{\circ}C$.

• Journal of Ginseng Research
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• 제24권2호
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• pp.64-67
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• 2000

본 연구에서는 홍삼에 함유되어 있는 총 phenolic화합물을 정량하기 위하여 Folin-Denis방법을 사용하였으며, 이 때 정량에 방해를 줄 수 있는 물질에 대해서 조사하였고 홍삼분말로부터 phenolic 화합물 추출조건을 조사하였다. 아미노산중에서는 tyrosine, cystein 및 tryptophan이 영향을 미쳤고, 여러 가지 유기산, 당류 및 ginsenosides는 거의 영향을 미치지 않았다. 비타민 중에서는 vitimln B$_{12}$, d-biotin, D-pantothenic acid, niacinamide, nicotinic acid, vitamine A palmitate, vitamine D$_3$ 등을 제외한 비타민은 발색에 영향을 미쳤다. 홍삼으로부터 phenolic 화합물을 추출하기 위한 추출조건은 60% ethanol을 사용해서 80。C에서 1-2시간씩 3회 정도 추출이 적당하다고 판단된다.

• 전기학회논문지
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• 제56권8호
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• pp.1424-1429
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• 2007

This paper presents a simple and reliable one-touch type multi-immunosensing lab-on-a-chip (LOC) detecting antibodies as multi-disease markers using electrochemical method suitable for a portable point-of-care system (POCS). The multi-stacked LOC consists of a PDMS space layer for liquids loading, a PDMS valve layer with 50 im in height for the membrane, a PDMS channel layer for the fluid paths, and a glass layer for multi electrodes. For the disposable immunoassay which needs sequential flow control of sample and buffer liquids according to the designed strategies, reliable and easy-controlled on-chip operation mechanisms without any electric power are necessary. The driving forces of sequential liquids transfer are the capillary attraction force and the pneumatic pressure generated by air bladder push. These passive fluid transport mechanisms are suitable for single-use LOC module. Prior to the application of detection of the antibody as a disease marker, the model experiments were performed with anti-DNP antibody and anti-biotin antibody as target analytes. The flow test results demonstrate that we can control the fluid flow easily by using the capillary stop valve and the PDMS check valves. By the model tests, we confirmed that the proposed LOC is easily applicable to the bioanalytic immunosensors using bioelectrocatalysis.

• Bulletin of the Korean Chemical Society
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• 제27권3호
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• pp.407-412
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• 2006

Bioluminescence single-site immunometric assay for methamphetamine (MA) using the native aequorin, a photoprotein, as a signal generator was developed for the first time. MA is a potent sympathomimetic amine with stimulant effects on the central nervous system. MA abuse induces hallucinations and, thus, may cause a serious social problem. The single-site immunometric MA assay was optimized and its dose-response behavior was examined. The dose-response curve shows that the detection limit is 1.1 ${\times}$ $10^{-10}$ M and a dynamic range is four orders of magnitude with 15 $\mu$g/mL BSA-MA conjugate and 1.0 ${\times}$ $10^{-8}$ M anti-MA antibody-biotin conjugate. In order to evaluate this assay, the structurally similar compounds, amphetamine, ephedrine, norephedrine, benzphetamine and N-4-(aminobutyl)methamphetamine were examined for their crossreactivity. None of these five compounds showed any cross-reactivity. Additionally, an artificial urine solution spiked with MA was analyzed by the MA assay, and the result of the analysis demonstrated the usefulness of the present assay for the determination of MA in urine.

• BMB Reports
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• 제30권6호
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• pp.403-409
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• 1997

Studies were undertaken to develop a competitive radioimmunoassay (RIA) and indirect antigen capture enzyme-linked immunosorbent assay (ELISA) for the determination of 5'-deoxy-5'-methylthioadenosine (MTA), which is formed from decarboxylated S-adenosylmethionine by spermidine and spermine synthase. Specific antiserum against MTA was raised in rabbits by immunization with MTA-BSA which was prepared by coupling BSA to oxidized MTA with periodate. Since MTA is oxidized easily to the sulfoxide, the sulfhydryl reagent, DTT. was added to the immunogen. For RIA, immunocomplexes were separated from free MTA by using ammonium sulfate precipitation. The antiserum showed almost no cross-reactivity with a variety of other nucleotides and riboses. But, the level of cross-reactivity of 5'-isobutylthioadenosine (SIBA) was high. These results showed the importance of hydrophobicity adjacent to the 5'-OH for determining antigenicity. The lower limit of detection by this assay was 100 fmol of MTA per tube. Using this assay. MTA levels were more easily and precisely determined in biological samples when compared with HPLC analysis. The RIA procedure is less time consuming. More than 24 analyses can be carried out in 2 h and required only a very small amount of sample ($20{\mu}l$ serum). In ELISA, biotin conjugated MTA-BSA was used as the labelled MTA. The sensitivity limit of this assay was lower than 100 pmol.

• 대한기계학회논문집A
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• 제28권9호
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• pp.1429-1434
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• 2004

Immunoassay is one of the important analytical methods for clinical diagnoses and biochemical studies, but needs a long time, troublesome procedures and expensive reagents. In this study, therefore, we propose the micro filter chip with microbeads for immunoassay, which has pillar structures. The advantage of the proposed micro filter chip is to use simple fabrication process and cheap materials. The mold was made by the photolithography technique with Si wafer and negative photoresist SU-8. The replica was made of PDMS, bonded on the pyrex glass. The micro filter chip consists of inlet channel, filter chamber and outlet channel. HBV (Hepatitius B virus) monoclonal antibody (Ag1) labeled with biotin were immobilized onto streptavidin coated beads of 30∼50 $\mu$m size. Fluorescein isothiocyanate (FITC)-labeled HBV monoclonal antibody (Ag8) was used to detect HBsAg (Hebatitis B virus surface Antigen), and fluorescence intensity was monitored by epi-fluorescence microscope. In this study, the immune response of less than 30 min was obtained with with the use of 100 $m\ell$ of sample.

• Biotechnology and Bioprocess Engineering:BBE
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• 제9권4호
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• pp.313-317
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• 2004

The nutritional requirements for Prevotella sp. 4PCCNB2 isolated from the rumen of a native goat in Korea and those of the ATCC 19189 strain isolated from the bovine rumen were investigated. The two strains grew well with ammonium sulfate as the sole added nitrogen source. However, neither a complex of amino acids nor casein hydrolysate effectively replaced ammonium sulfate. Biotin, p-aminobenzoic acid, and vitamin $B_12$ were essential to culture the ATCC 19189 strain. Unlike the ATCC 19189 strain, however, $B_12$ was only stimulatory for the growth of the 4PCCNB2 strain. The 4PCCNB2 strain grew well in the basal medium without an individual acid such as acetic acid or valeric acid. In contrast, either acetic or valeric acid was absolutely required for the growth of the ATCC 19189 strain.