• 제목/요약/키워드: copy numbers

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벼 조직배양 기간에 따른 retrotransposon(Tos17)의 활성에 관한 연구 (The Studies of Activity of Retrotransposon(Tos17) according to Tissue Culture Periods in Rice(Oryza sativa L.))

  • 양희은;방일란;신영범;이병진;홍순관
    • 한국자원식물학회지
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    • 제20권5호
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    • pp.389-397
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    • 2007
  • 벼의 캘러스를 대상으로 Tos17 활성 증가를 이용하여 새로운 돌연변이체의 유발 및 선발에 대한 연구는 많이 시도되어 왔다. 일품벼에서 배양된 캘러스를 이용하여 배양기간 및 배양조건에 따른 retrotransposon(Tos17)의 활성화를 유도함을 일차적인 목적으로 하고, 이에 따른 재분화 개체를 통하여 다양한 돌연변이체($M_1$)를 얻음과 동시에 세대의 진전에 따른 돌연변이체($M_2$, $M_3$)가 나타내는 표현형과 Tos17과의 연관성을 확인하여 특정 유전자의 cloning을 향후 목적으로 하고 있다. 1. 본 연구에서는 배양기간에 따라 총 371개체의 $M_1$ 돌연변이체를 얻었다. 2. 각각의 배양기간 단계별 재분화 식물체에서 얻어진 Tos17의 활성정도를 나타내는 Southern blot의 결과 normal 즉, 기본 식물인 일품벼는 5개의 copy수를 가지고 있는 것을 알 수 있었으며, 1개월 7개, 2개월 8개, 3개월 9.5개, 5개월 12개, 6개월 6개, 7개월 13.5개, 8개월 17.5개의 band를 확인할 수 있었다. 3. Tos17의 Southern blot의 결과, $3{\sim}5$개월의 배양기간이 경과해야만 기본 copy의 2배수로 Tos17이 활성화됨을 알 수 있었다. 향후 $M_1$돌연변이체의 세대진전을 통하여 $M_2$, $M_3$ 세대 등의 다양한 재조합 개체의 확보 및 분석이 중요하다. 또한 다양한 형태로 원하는 돌연변이를 선발하는 screening 방법을 개발하는 것이 시급하고 중요한 과제이다.

갓 (Brassica juncea) 품종구분을 위한 ITS 영역 및 MITE Family 정보를 이용한 분자표지 개발 (Development of molecular markers for varietal identification of Brassica juncea on the basis of the polymorphic sequence of ITS regions and MITE families)

  • 양기웅;이고은;아리프 하산 칸 로빈;정남희;이용혁;박종인;김회택;정미영;노일섭
    • 원예과학기술지
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    • 제34권2호
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    • pp.305-313
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    • 2016
  • 갓(Brassica juncea; 2n = 4x = 36, AABB genome, 1,068Mb)은 U's triangle의 배추와 흑겨자 사이의 복이배체 작물로 구분한다. 본 연구는 갓 15 품종의 ribosomal DNA ITS 영역과 MITE를 이용하여 갓의 유연관계 및 품종구분 분자표지를 확인하였다. Ribosomal DNA ITS 영역은 종 및 품종의 유연관계를 알아보는 연구로 많이 사용되고 있어서, 이를 이용하여 갓 15 품종의 유연관계를 알아보았다. 또한, MITE는 매우 많은 copy 수를 가지고 있고, 유전적으로 안정적이기 때문에 유전체 및 진화 연구에 매우 적합한 재료이다. MITE를 이용한 갓의 품종구분 분자표지를 확인하기 위해 MITE super-families 중 Stowaway(BraSto) 관련 70점, Tourist(BraTo) 관련 79점, hAT(BrahAT) 관련 6점, Mutator(BraMu) 관련 5점으로 품종구분 표지를 알아보았다. 총 160점의 분자표지 중 32점이 갓 15 품종에서 뚜렷한 다형성을 보였다. 특히, 흑갓은 표현형뿐만 아니라 유전자형도 매우 다르게 나타났다. 또한 8점의 MITE 분자표지를 활용하여 47점의 유전자원에서 다형성 및 품종구분 표지로의 활용 가능성을 확인하였다. 이러한 다형성 표지들은 갓의 품종구분 및 품종 보호에 매우 유용하게 사용할 수 있을 것이라 기대한다.

동해 울릉분지 가스 하이드레이트 매장 지역의 메탄산화 미생물 군집 조성 및 분포 (Microbial Community Composition Associated with Anaerobic Oxidation of Methane in Gas Hydrate-Bearing Sediments in the Ulleung Basin, East Sea)

  • 조혜연;김성한;신경훈;박장준;현정호
    • 한국해양학회지:바다
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    • 제20권1호
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    • pp.53-62
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    • 2015
  • 동해 울릉 분지 내 메탄 하이드레이트가 매장된 지역에서 혐기적 메탄 산화와 연관된 미생물 군집 특성을 이해하기 위해 메탄 누출이 있는 대륙사면 정점(UBGH2-3)과 메탄 누출이 없는 분지 정점(UBGH2-10)에서, (1) 퇴적물의 지화학적 성분 및 황산염 환원율을 측정하였으며, (2) 기능성 유전자 분석을 통해 혐기적 메탄 산화 미생물 및 황산염 환원 미생물 군집의 정량 및 다양성 분석을 수행하여 그 결과를 비교하였다. 황산염-메탄 전이지대(sulfate and methane transition zone, SMTZ)는 UBGH2-3에서 0.5~1.5 mbsf (meters below seafloor), 그리고 UBGH2-10에서는 6~7 mbsf에 분포하는 것으로 나타났다. 두 지역의 SMTZ에서 측정된 황산염 환원율은 UBGH2-3의 1.15 mbsf에서 $1.82nmol\;cm^{-3}d^{-1}$으로 나타났고, UBGH2-10의 SMTZ에서 황산염 환원율은 $4.29nmol\;cm^{-3}d^{-1}$으로 높은 값을 보였다. 총 미생물 16S rRNA gene과 기능성 유전자인 mcrA (methyl coenzyme M reductase subunit A) 및 dsrA (dissimilatory sulfite reductase subunit A)의 정량 PCR 결과 두 정점의 SMTZ 부근에서 상대적으로 높게 검출되었다. 그러나 UBGH2-10지역에서 mcrA 유전자는 SMTZ 아래인 9.8 bmsf에서 가장 높게 검출되었다. mcrA 유전자의 다양성 분석 결과 두 지역의 SMTZ와 그 아래 퇴적층에서 혐기성 메탄산화 고세균(ANME: Anaerobic MEthanothoph) 군집인 ANME-1이 우점하였다. 한편, ANME-2 군집은 메탄 누출이 발생하는 UBGH2-3지역의 2.2 mbsf 층에서만 관찰되었고, 더불어 dsr 유전자 다양성 분석 결과 ANME-2와 컨소시엄을 이루는 Desulfosarcina-Desulfococcus (DSS) 이 나타났다. 본 연구 결과, ANME-1과 ANME-2은 동해 심부 퇴적 환경에서 혐기적 메탄 산화 및 생성 과정에 관여하는 중요한 고세균 군집으로 사료되며, 또한 ANME-2/DSS 컨소시엄은 메탄이 누출되는 지역인 UBGH2-3과 같이 혐기적 메탄 산화가 활발한 곳에서 메탄 거동을 조절하는 중요한 미생물 그룹으로 인식된다.

Effects of Protein Level and Mangosteen Peel Pellets (Mago-pel) in Concentrate Diets on Rumen Fermentation and Milk Production in Lactating Dairy Crossbreds

  • Norrapoke, T.;Wanapat, M.;Wanapat, S.
    • Asian-Australasian Journal of Animal Sciences
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    • 제25권7호
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    • pp.971-979
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    • 2012
  • Four, lactating dairy crossbreds ($50%{\times}50%$ Holstein Friesian${\times}$Native Zebu cattle) were randomly assigned according to a $2{\times}2$ factorial arrangement (two protein levels and two levels of mangosteen peel pellets (Mago-pel)) in a $4{\times}4$ Latin square design to receive four dietary treatments. All cows received concentrate at a proportion of 1 kg concentrate per 2 kg of milk yield, and urea-treated 5% rice straw (UTRS) was given ad libitum. It was found that total dry matter intakes, nutrient digestibility, ruminal pH and $NH_3$-N concentrations were not affected (p>0.05) by treatments. Concentrations of ruminal pH and $NH_3$-N were not affected by dietary treatments although the concentration of BUN varied significantly (p<0.05) between protein levels (p<0.05). The populations of rumen bacteria and fungal zoospores did not differ among treatments (p>0.05); however, the population of protozoa was decreased (p<0.05) when cows received Mago-pel supplementation. The composition of the population of bacteria, identified by real-time PCR technique, including total bacteria, methanogens, Fibrobacter succinogenes and Ruminococcus albus was similar (p>0.05) among dietary treatments (p>0.05); however, copy numbers of Ruminococcus flavefaciens was increased when protein level increased (p<0.05). Microbial protein synthesis, in terms of both quantity and efficiency, was enriched by Mago-pel supplementation. Milk yield was greatest in cows fed UTRS based diets with concentrate containing protein at 16% CP with Mago-pel, but were lowest without Mago-pel (p<0.05). In addition, protein level and supplementation of Mago-pel did not affect (p>0.05) milk composition except solids-not-fat which was higher in cows fed the diet with 19% CP. Therefore, feeding a concentrate containing 16% CP together with 300 g/hd/d Mago-pel supplementation results in changes in rumen fermentation and microbial population and improvements in milk production in lactating dairy crossbreds fed on UTRS.

Production of ʟ-Theanine Using Escherichia coli Whole-Cell Overexpressing γ-Glutamylmethylamide Synthetase with Baker's Yeast

  • Yang, Soo-Yeon;Han, Yeong-Hoon;Park, Ye-Lim;Park, Jun-Young;No, So-young;Jeong, Daham;Park, Saerom;Park, Hyung Yeon;Kim, Wooseong;Seo, Seung-Oh;Yang, Yung-Hun
    • Journal of Microbiology and Biotechnology
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    • 제30권5호
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    • pp.785-792
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    • 2020
  • ʟ-Theanine, found in green tea leaves has been shown to positively affect immunity and relaxation in humans. There have been many attempts to produce ʟ-theanine through enzymatic synthesis to overcome the limitations of traditional methods. Among the many genes coding for enzymes in the ʟ-theanine biosynthesis, glutamylmethylamide synthetase (GMAS) exhibits the greatest possibility of producing large amounts of production. Thus, GMAS from Methylovorus mays No. 9 was overexpressed in several strains including vectors with different copy numbers. BW25113(DE3) cells containing the pET24ma::gmas was selected for strains. The optimal temperature, pH, and metal ion concentration were 50℃, 7, and 5 mM MnCl2, respectively. Additionally, ATP was found to be an important factor for producing high concentration of ʟ-theanine so several strains were tested during the reaction for ATP regeneration. Baker's yeast was found to decrease the demand for ATP most effectively. Addition of potassium phosphate source was demonstrated by producing 4-fold higher ʟ-theanine. To enhance the conversion yield, GMAS was additionally overexpressed in the system. A maximum of 198 mM ʟ-theanine was produced with 16.5 mmol/l/h productivity. The whole-cell reaction involving GMAS has greatest potential for scale-up production of ʟ-theanine.

유기농 옥수수밭에서 경운이 토양 유기물 함량 및 미생물군집에 미치는 영향 (Effects of Tillage on Organic Matters and Microbial Communities in Organically Cultivated Corn Field Soils)

  • 안달래;안난희;김다혜;한병학;유재홍;박인철;안재형
    • 한국환경농학회지
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    • 제39권1호
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    • pp.65-74
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    • 2020
  • BACKGROUND: Soil carbon sequestration has been investigated for a long time because of its potential to mitigate the greenhouse effect. No- or reduced tillage, crop rotations, or cover crops have been investigated and practiced to sequester carbon in soils but the roles of soil biota, particularly microorganisms, have been mostly ignored although they affect the amount and stability of soil organic matters. METHODS AND RESULTS: In this study we analyzed the organic matter and microbial community in organically cultivated corn field soils where no-tillage (NT) or conventional tillage (CT) had been practiced for about three years. The amounts of organic matter and recalcitrant carbon pool were 18.3 g/kg dry soil and 4.1 g C/kg dry soil, respectively in NT soils, while they were 12.4 and 2.5, respectively in CT soils. The amounts of RNA and DNA, and the copy numbers of bacterial 16S rRNA genes and fungal ITS sequences were higher in NT soils than in CT soils. No-tillage treatment increased the diversities of soil bacterial and fungal communities and clearly shifted the bacterial and fungal community structures. In NT soils the relative abundances of bacterial phyla known as copiotrophs, Betaproteobacteria and Bacteroidetes, increased while those known as oligotrophs, Acidobacteria and Verrucomicrobia, decreased compared to CT soils. The relative abundance of a fungal phylum, Glomeromycota, whose members are known as arbuscular mycorrhizal fungi, was about two time higher in NT soils than in CT soils, suggesting that the higher amount of organic matter in NT soils is related to its abundance. CONCLUSION: This study shows that no-tillage treatment greatly affects soil microbial abundance and community structure, which may affect the amount and stability of soil organic matter.

The Uneven Distribution of Mating Type Genes in Natural and Cultivated Truffle Orchards Contributes to the Fructification of Tuber indicum

  • Li, Qiang;Fu, Yu;Sun, Qun;Sourzat, Pierre;Yang, Mei;Liu, Chengyi;Tan, Hao;Ye, Lei;Zou, Jie;Wu, Chenguang;Zhang, Bo;Li, Xiaolin
    • Mycobiology
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    • 제46권1호
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    • pp.64-71
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    • 2018
  • The aim of this study was to investigate the pattern of distribution of mating type (MAT) genes of Tuber indicum in ectomycorhizosphere soils from natural T. indicum-producing areas and cultivated truffle orchards and ascocarp samples from different regions. Quantitative real-time PCR and multiplex PCR were used to weight the copy numbers of MAT1-1-1 and MAT1-2-1 in natural truffle soils and cultivated orchard soils. The effect of limestone on the pattern of truffle MAT genes and the correlation between soil properties and the proportion of MAT genes were also assessed. These results indicated that an uneven and nonrandom distribution of MAT genes was common in truffle-producing areas, cultivated truffle orchards, and ascocarps gleba. The competition between the two mating type genes and the expansion of unbalanced distribution was found to be closely related to truffle fructification. Limestone treatments failed to alter the proportion of the two mating type genes in the soil. The content of available phosphorus in soil was significantly correlated with the value of MAT1-1-1/MAT1-2-1 in cultivated and natural ectomycorhizosphere soils. The application of real-time quantitative PCR can provide reference for monitoring the dynamic changes of mating type genes in soil. This study investigates the distributional pattern of T. indicum MAT genes in the ectomycorhizosphere soil and ascocarp gleba from different regions, which may provide a foundation for the cultivation of T. indicum.

Evaluation of liquid and powdered forms of polyclonal antibody preparation against Streptococcus bovis and Fusobacterium necrophorum in cattle adapted or not adapted to highly fermentable carbohydrate diets

  • Cassiano, Eduardo Cuellar Orlandi;Perna, Flavio Junior;Barros, Tarley Araujo;Marino, Carolina Tobias;Pacheco, Rodrigo Dias Lauritano;Ferreira, Fernanda Altieri;Millen, Danilo Domingues;Martins, Mauricio Furlan;Pugine, Silvana Marina Piccoli;de Melo, Mariza Pires;Beauchemin, Karen Ann;Meyer, Paula Marques;Arrigoni, Mario de Beni;Rodrigues, Paulo Henrique Mazza
    • Animal Bioscience
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    • 제34권1호
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    • pp.74-84
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    • 2021
  • Objective: Feed additives that modify rumen fermentation can be used to prevent metabolic disturbances such as acidosis and optimize beef cattle production. The study evaluated the effects of liquid and powdered forms of polyclonal antibody preparation (PAP) against Streptococcus bovis and Fusobacterium necrophorum on rumen fermentation parameters in ruminally cannulated non-lactating dairy cows that were adapted or unadapted to a high concentrate diet. Methods: A double 3×3 Latin square design was used with three PAP treatments (control, powdered, and liquid PAP) and two adaptation protocols (adapted, unadapted; applied to the square). Adapted animals were transitioned for 2 weeks from an all-forage to an 80% concentrate diet, while unadapted animals were switched abruptly. Results: Interactions between sampling time and adaptation were observed; 12 h after feeding, the adapted group had lower ruminal pH and greater total short chain fatty acid concentrations than the unadapted group, while the opposite was observed after 24 h. Acetate:propionate ratio, molar proportion of butyrate and ammonia nitrogen concentration were generally greater in adapted than unadapted cattle up to 36 h after feeding. Adaptation promoted 3.5 times the number of Entodinium protozoa but copy numbers of Streptococcus bovis and Fibrobacter succinogens genes in rumen fluid were not affected. However, neither liquid nor powdered forms of PAP altered rumen acidosis variables in adapted or unadapted animals. Conclusion: Adaptation of cattle to highly fermentable carbohydrate diets promoted a more stable ruminal environment, but PAP was not effective in this study in which no animal experienced acute or sub-acute rumen acidosis.

공주시 무인민원발급기 이용 활성화를 위한 신분 확인 확대 방안 (Expansion of Personal Identification to Vitalize the Use of Unmanned Civil Service Issuing Machines for Gongju-si)

  • 김의석;정한민
    • 한국정보통신학회:학술대회논문집
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    • 한국정보통신학회 2021년도 추계학술대회
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    • pp.156-158
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    • 2021
  • 초기 지방자치센터는 민원인으로부터 민원을 접수하여 수기로 직접 해당 문건을 작성하여 민원을 발급하였으며, 컴퓨터, 프린터, 팩스가 등장한 이후에는 복사를 통해 민원 업무를 처리하였다. 2010년 이후에는 전자 정부가 출범하면서 컴퓨터에 민원 요청 내용을 입력하고 본인 확인 후에 직접 출력하는 방식이 자리를 잡게 되었다. 또한, 무인 행정 서비스가 도입되어 다양한 장소에서 민원 서비스를 제공하면서 민원인의 만족도가 점차 높아지고 있다. 그중 하나의 서비스가 무인민원발급기이며 주민등록등본을 포함한 20여 가지의 민원 서류들을 민원인이 직접 발급받을 수 있도록 하고 있다. 물론, 직접 민원 창구에서 요구하던 방식을 벗어나 직접 기기를 다루다 보니 최신 기술에 익숙하지 않았을 때 도움이 추가로 필요한 경우가 생기기도 한다. 그렇지만, 무인민원발급기는 업무 종료 후에도 주민자치센터나 시청을 방문하여 민원 처리를 할 수 있다는 점에서 시간의 제약을 덜 받는, 만족도가 높은 방안이다. 현재 주요한 문제는 발급 종류에 따라 주민등록번호뿐만 아니라 지문 인식도 요구하는 경우가 있어, 민원 처리의 불편이 가중되고 외면받는 것이다. 이러한 문제를 해결하고자, 본 연구는 개인 신분 확대를 통해 무인민원발급기의 지문 인식 방법을 개선하는 방법을 연구하고자 한다.

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Detection of Campylobacter jejuni in food and poultry visors using immunomagnetic separation and microtitre hybridization

  • Simard, Ronald-E.
    • 한국어업기술학회:학술대회논문집
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    • 한국어업기술학회 2000년도 춘계수산관련학회 공동학술대회발표요지집
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    • pp.71-73
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    • 2000
  • Campylobacter jejuni is most frequently identified cause of cause of acute diarrhoeal infections in developeed countries, exceeding rates of illness caused by both salmonella and shigilla(Skirrow, 1990 ; Lior 1994). Previous studies on campylobacter jejuni contamination of commercial broiler carcasses in u.s.(Stern, 1992). Most cases of the disease result from indirect transmission of Campylobactor from animals via milk, water and meat. In addition to Campylobactor jejuni. the closely relates species Campylobactor coli and Campylobactor lari have also been implicated as agents of gastroenteritis in humans. Campylobactor coli represented only approximately 3% of the Campylobactor isolates from patients with Campylobactor enteritis(Griffiths and Park, 1990) whereas Campylobactor coli is mainly isolated from pork(Lmmerding et al., 1988). Campylobactor jejuni has also been isolated from cases of bacteremia, appendicitis and, recently, has been associated with Guillai-Barre syndrome(Allos and Blaser, 1994; von Wulffen et al., 1994; Phillips, 1995). Studies in volunteers indicated that the infectious dose for Campylobactor jejuni is low(about 500 organisms)(Robinson, 1981). The methods traditionally used to detect Campylobactor ssp. in food require at least two days of incubation in an enrichment broth followed by plating and two days of incubation on complex culture media containing many antibiotics(Goossens and Butzler, 1992). Finnaly, several biochemical tests must be done to confirm the indentification at the species level. Therfore, sensitive and specific methods for the detection of small numbers of Campylobactor cells in food are needed. Polymerase chain reaction(PCR) assays targeting specific DNA sequences have been developed for the detection of Campylobactor(Giesendorf and Quint, 1995; Hemandex et al., 1995; Winter and Slavidk, 1995). In most cases, a short enrichment step is needed to enhance the sensitivity of the assay prior to detection by PCR as the number of bacteria in the food products is low in comparison with those found in dinical samples, and because the complex composition of food matrices can hinder the PCR and lower its sensitivity. However, these PCR systems are technically demanding to carry out and cumbersome when processing a large number of samples simutaneously. In this paper, an immunomagnetic method to concentrate Campylobactor cells present in food or clinical samples after an enrichment step is described. To detect specifically the thermophilic Campylobactor. a monoclonal antibody was adsorbed on the surface of the magnetic beads which react against a major porin of 45kDa present on the surface of the cells(Huyer et al., 1986). After this partial purification and concentration step, detection of bound cells was achieved using a simple, inexpensive microtitre plate-based hybridization system. We examined two alternative detection systems, one specific for thermophilic Campylobactor based on the detection of 23S rRNA using an immobilized DNA probe. The second system is less specific but more sensitive because of the high copy number of the rRNA present in bacterial cell($10^3-10^4$). By using specific immunomagnetic beads against thermophilic Campylobactor, it was possible to concentrate these cells from a heterogeneous media and obtain highly specific hybridization reactions with good sensitivity. There are several advantages in using microtitre plates instead of filter membranes or other matrices for hybridization techniques. Microtitre plates are much easier to handle than filter membranes during the adsorption, washing, hybridization and detection steps, and their use faciilitates the simultanuous analysis of multiple sample. Here we report on the use of a very simple detection procedure based on a monoclonal anti-RNA-DNA hybrid antibody(Fliss et al., 1999) for detection of the RNA-DNA hybrids formed in the wells.

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