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Pectobacterium carotovorum의 검출을 위한 PCR 진단법의 개발
노지나,유미선,박동석,김정구,윤병수,No, Ji-Na,Yoo, Mi-Sun,Park, Dong-Suk,Kim, Jeong-Gu,Yoon, Byoung-Su 한국미생물학회 2009 미생물학회지 Vol.45 No.4
Pectobacterium carotovorum은 배추를 비롯한 광범위한 식물체에 무름병을 일으키는 병원성 세균으로, 무름병의 효율적인 방제를 위하여 신속한 병원체의 진단이 요구되고 있다. 따라서 본 연구에서는 높은 특이성으로 다양한 진단에 적용되고 있는 PCR법을 이용하여 Pectobacterium carotovorum을 높은 정확성과 민감성으로 검출할 수 있는 진단법을 개발하고자 하였다. 먼저 P. carotovorum에 특이성이 있다고 보고된 다양한 특이 primer들을 비교하여 가장 특이성이 높은 primer쌍들을 선별하였으며, 최종 선발된 특이 primer쌍은 ERB_3F (5'-TGCGACACCTCCTCATCACG-3'), ERB_3R (5'-CTTATCACGCTGTAACCAGC-3')로 나타났다. 이들을 사용한 PCR 검출법은 $58^{\circ}C$의 annealing 온도, 15 mM $MgCl_2$ 농도 등으로 최적화되었으며, 최적조건에서 P. carotovorum 특이 PCR 진단법은 10 pg, 즉 $2\times10^3$ copies의 병원균 유전자를 검출할 수 있는 우수한 민감성을 보였다. 또한 배양된 균주가 아닌 현장 시료에서 본 P. carotovorum 검출법을 시험해 본 결과 이 검사법은 병원균 배양없이 현장에도 직접 적용할 수 있음을 입증하였다. 따라서 본 연구를 통해 확립된 P. carotovorum 특이 PCR 진단법은 해당 병원균을 신속하게 진단하는데 유용하게 사용될 수 있을 것이라 기대한다. A new PCR method was developed to detect Pectobacterium carotovorum which is the causative agent of soft rot in Brassica pekinensis. A specific detection primer set based on Lytic murein transglycolase gene was designed and evaluated. Using ERB3_F (5'-TGC GAC ACC TCC TCA TCA CG-3') and ERB3_R (5'-CTT ATC ACG CTG TAA CCA GC-3') primers, 437 nucleotides long fragment was specifically amplified. The amplified products were observed in 52 out of 55 strains of P. carotovorum or Pectobacterium carotovorum subsp. carotovorum. On the other hand, no amplification was observed in 8 organisms including Chinese cabbage and potato. The optimal PCR condition for the ERB3_F/ERB3_R primer set was $58^{\circ}C$ for annealing and 15 mM for $MgCl_2$. With serially diluted templates, the specific PCR sensitivitie limit was $2\times10^3$ copies. Also, this method can be applied not only to DNA but also to field samples. This PCR method may be expected to be useful for specific detection of P. carotovorum.
디포스핀류 배위자를 함유한 백금(II) 착체의 합성과 항암효과
노영수,노지 마사히데,Noh, Young-Soo,Masahide, Noji 한국약제학회 1990 Journal of Pharmaceutical Investigation Vol.20 No.4
New antitumor-active pt(II) complexes of trans-l-diamine cyclohexane containing diphosphines as a leaving group were synthesized. The structures of the pt(II) complexes were determined by analyzing the infrared and $^{31}P-nuclear$ magnetic resonance spectra. Antitumor activities of the pt(II) complexes were tested against murine leukemia $L_{l210}$ according to the protocol of the National Cancer Institute. All the pt(II) complexes Synthesized were antitumor-active. In particular, water-soluble $[pt(trans-l-dach) (DPPP)](NO_3)_2$ exhibited excellent antitumor activity, giving T/C % values of 341 and 356 respectively, each with four cured mice out of six at a dose of 25 mg/kg. These pt(II) complexes are considered to be worthy of further development.
시지야 이논게 노니,서명지 한국식품과학회 2023 식품과학과 산업 Vol.56 No.1
Lactic acid bacteria (LAB) are well-known for their role in the food, agriculture, and health industries. The health benefits of LAB include anti-inflammatory and antioxidant protection, immunomodulatory effects, anti-obesity effects, enhanced bioavailability of bioactive compounds, and antimicrobial properties. Many LAB are well-known probiotics and certain species have been identified as carotenoid-producers, giving them a new identity as potential postbiotic sources of carotenoids, in addition to the benefits that LAB already possess. Although the most abundant microbial carotenoids have C40 backbones, carotenogenic genes in LAB are specific to C30 carotenoid synthesis. This review discusses the production and optimization of identified C30 carotenoids by LAB and the current status of C30 carotenoid-producing LAB, to establish their potential in postbiotic development. The absorption of carotenoids in the gut is also explored to better understand the applications of viable LAB-carotenoid postbiotics in fermented food products and probiotic supplements.
Israeli Acute Paralysis Virus (IAPV) 검출을 위한 Semi-nested PCR 법의 개발
노지나(Ji-Na No),Nguyen Van Phu,유미선(Mi-Sun Yoo),윤병수(Byoung-Su Yoon) 한국양봉학회 2010 韓國養蜂學會誌 Vol.25 No.2
A new semi-nested PCR method was developed to detect Israeli acute paralysis virus (IAPV) which is the causative virus of colony collapse disorder (CCD) in honeybee. To amplify IAPV structural protein gene, the sequences of IAPV including accession No. ER219380, EU436455, EU436456, EU224279, EU224280, EU436423, EU218534 in GenBank were analysed. Two pairs of IAPV specific primer sets named IAPV_F1/2, IAPV_R1 outer and IAPV_F1/2, IAPV_R2 inner were designed and evaluated. As results, the unique 284bp DNA fragment was successfully amplified with high sensitivity using IAPV specific semi-nested PCR method. From only three copies of IAPV-template, the IAPV specific seminested PCR showed positive amplification. From field sample its sensitivity was also verified. The IAPV semi-nested PCR method would be applicable to the early detection of IAPV and the confirmation test for existence of few IAPVs in nature.
Loop-mediated Isothermal Amplification (LAMP)법을 이용한 Chronic Bee Paralysis Virus (CBPV)의 신속 진단법 개발
노지나(Ji-Na No),유미선(Mi-Sun Yoo),Van Phu Nugyen,윤병수(Byoung Su Yoon) 한국양봉학회 2010 韓國養蜂學會誌 Vol.25 No.4
Chronic bee paralysis virus (CBPV) is the causal agent of chronic paralysis in honeybee colonies. A loop-mediated isothermal amplification (LAMP) assay allows one-step detection of gene amplification by simple reaction and requires only a simple incubator, such as water bath providing a constant temperature. In this study, CBPV-LAMP was developed for detection of CBPV. The four CBPVspecific primers named CBPV-LAMP-F3/B3/FIP/BIP were designed. CBPV was successfully amplified under isothermal conditions at about 54°C within 30 minutes and CBPV-LAMP could be detect untill 1×10⁴ copies. Especially, developed method, CBPV-LAMP was direct detection analysis by the naked eye without electrophoresis using SYBR Green I. This method provides an important diagnostic tool for the detection of CBPV infection.
IAPV내의 capsid protein gene의 검출을 위한 RT-LAMP 법의 개발
노지나(Ji-Na No),이보람(Bo-Ram Lee),유미선(Mi-Sun Yoo),윤병수(Byoung Su Yoon) 한국양봉학회 2011 韓國養蜂學會誌 Vol.26 No.2
Israel acute paralysis virus (IAPV) was reported as a virus of colony collapse disorder (CCD) in honeybee. A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay allows one-step detection of gene amplification by simple analysis and requires only a simple incubator, such as water bath providing a constant temperature. RT-LAMP method was applied for simple and rapid detection of IAPV. This method was named IAPV-RT-LAMP. The four primers named IAPV-VP2-F3/B3/FIP/BIP were designed for specific amplification of VP2 capsid protein gene in IAPV. Especially, total diagnosis time was reduced until 40 miuntes including reverse transcription reaction from RNA stage. After reaction, LAMP products added fluorescent dye, SYBR Green I and GeneFinder™ were observed with naked eye under daylight without electrophoresis. IAPV-RT-LAMP is expected to be an importance diagnostic tool for detection of IAPV both in laboratory and in field.