• 제목/요약/키워드: Reverse transcription

검색결과 1,335건 처리시간 0.028초

Abundance and expression of denitrifying genes (narG, nirS, norB, and nosZ) in sediments of wastewater stabilizing constructed wetlands

  • Chon, Kyongmi;Cho, Jaeweon
    • Environmental Engineering Research
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    • 제20권1호
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    • pp.51-57
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    • 2015
  • As expected, the expression of denitrifying genes in a Typha wetland (relatively stagnant compared to other ponds), showing higher nitrogen removal efficiency in summer, was affected by temperature. The abundance and gene transcripts of nitrate reductase (narG), nitrite reductase (nirS), nitric oxide reductase (norB), and nitrous oxide reductase (nosZ) genes in seasonal sediment samples taken from the Acorus and Typha ponds of free surface flow constructed wetlands were investigated using quantitative polymerase chain reaction (Q-PCR) and quantitative reverse transcription PCR (Q-RT-PCR). Denitrifying gene copy numbers ($10^5-10^8$ genes $g^{-1}$ sediment) were found to be higher than transcript numbers-($10^3-10^7$ transcripts $g^{-1}$ sediment) of the Acorus and Typha ponds, in both seasons. Transcript numbers of the four functional genes were significantly higher for Typha sediments, in the warm than in the cold season, potentially indicating greater bacterial activity, during the relatively warm season than the cold season. In contrast, copy numbers and expression of denitrifying genes of Acorus did not provide a strong correlation between the different seasons.

해녀콩(Canavalia lineata THUNB. DC.) 추출물의 멜라닌 생성 억제 효과 (Anti-melanogenesis Effect of Canavalia lineata Extract)

  • 부희정;류기중;이선주
    • 대한화장품학회지
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    • 제30권4호
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    • pp.485-489
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    • 2004
  • 피부에서 melanin은 자외선 차단의 주요한 역할을 한다. Tyrosinase는 멜라닌 생합성과정에서 초기 단계에 관여하는 중요한 효소로서 이것의 조절을 통한 피부 멜라닌화 억제에 관해 많은 연구가 되어져왔다. 본 연구에서는 해녀콩 추출물에서 mushroom tyrosinase 활성억제, B16F10 melanoma 세포를 이용한 dopa oxidase 활성억제 및 멜라닌 합성 억제 효과를 확인하였다. Tyrosinase mRNA 발현에서의 억제 효과를 확인하기 위하여 RT-PCR을 이용하였으며, $CHCI_3$ 층에서 분리해낸 A 분획에서 tyrosinase mRNA 발현을 억제시킴을 확인하였다.

Identification of Genes Induced by Low Temperature in Rice

  • Choi, Kyong-Hee;Choi, Hack-Sun;Lee, Choon-Hwan;Kwon, Young-Myung;Rhew, Tae-Hyong
    • BMB Reports
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    • 제30권4호
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    • pp.292-295
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    • 1997
  • Exposure of seedling of rice (Oriza sativa cv.Dongin) to cold stress ($6^{circ}C$, 7day) induced differential gene expression. Differentially expressed polyadenylated RNA induced by low temperature were isolated and identified from the leaves of rice (Oriza sativa cv.Dongin) seedling by using the technique, differential display of reverse transcription through polymerase chain reaction (DDRT-PCR). Four bands of cDNAs were differentially displayed on the PAGE gel through DDRT-PCR, and among them three bands were those of overexpressed genes while one band was of an underexpressed gene One of the overexpressed cDNA was characterized. The size of the DDRT-PCR product was found to be about 200 bp. The sequence of the cloned DNA was compared with those of GenBank through a BLAST E-Mail server, and it was found to have no homologies in the nucleotide sequence with that of any known DNA: therefore, it was designated as RC101 The expression of the cold-stress induced-gene, RC101, was sustained with Northern Blot analysis by using the cloned DDRT-PCR product as a probe.

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Reverse Transcription Polymerase Chain Reaction-based System for Simultaneous Detection of Multiple Lily-infecting Viruses

  • Kwon, Ji Yeon;Ryu, Ki Hyun;Choi, Sun Hee
    • The Plant Pathology Journal
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    • 제29권3호
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    • pp.338-343
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    • 2013
  • A detection system based on a multiplex reverse transcription (RT) polymerase chain reaction (PCR) was developed to simultaneously identify multiple viruses in the lily plant. The most common viruses infecting lily plants are the cucumber mosaic virus (CMV), lily mottle virus (LMoV), lily symptomless virus (LSV). Leaf samples were collected at lily-cultivation facilities located in the Kangwon province of Korea and used to evaluate the detection system. Simplex and multiplex RT-PCR were performed using virus-specific primers to detect single- or mixed viral infections in lily plants. Our results demonstrate the selective detection of 3 different viruses (CMV, LMoV and LSV) by using specific primers as well as the potential of simultaneously detecting 2 or 3 different viruses in lily plants with mixed infections. Three sets of primers for each target virus, and one set of internal control primers were used to evaluate the detection system for efficiency, reliability, and reproducibility.

Mcl-PHAs Produced by Pseudomonas sp. Gl01 Using Fed-Batch Cultivation with Waste Rapeseed Oil as Carbon Source

  • Mozejko, Justyna;Wilke, Andreas;Przybylek, Grzegorz;Ciesielski, Slawomir
    • Journal of Microbiology and Biotechnology
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    • 제22권3호
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    • pp.371-377
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    • 2012
  • The present study describes medium-chain-length polyhydroxyalkanoates (mcl-PHAs) production by the Pseudomonas Gl01 strain isolated from mixed microbial communities utilized for PHAs synthesis. A two-step fed-batch fermentation was conducted with glucose and waste rapeseed oil as the main carbon source for obtaining cell growth and mcl-PHAs accumulation, respectively. The results show that the Pseudomonas Gl01 strain is capable of growing and accumulating mcl-PHAs using a waste oily carbon source. The biomass value reached 3.0 g/l of CDW with 20% of PHAs content within 48 h of cultivation. The polymer was purified from lyophilized cells and analyzed by gas chromatography (GC). The results revealed that the monomeric composition of the obtained polyesters depended on the available substrate. When glucose was used in the growth phase, 3-hydroxyundecanoate and 3-hydroxydodecanoate were found in the polymer composition, whereas in the PHAs-accumulating stage, the Pseudomonas Gl01 strain synthesized mcl-PHAs consisting mainly of 3-hydroxyoctanoate and 3-hydroxydecanoate. The transcriptional analysis using reverse-transcription real-time PCR reaction revealed that the phaC1 gene could be transcribed simultaneously to the phaZ gene.

Detection of Marine Birnavirus (MBV) from Rockfish Sebastes schlegeli Using Reverse Transcription and Nested PCR

  • Joh, Seong-Joon;Kim, Doo-Won;Kim, Jeong-Ho;Heo, Gang-Joon
    • Journal of Microbiology
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    • 제38권4호
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    • pp.260-264
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    • 2000
  • Reverse transcription (RT)-PCR and nested PCR methods (2-step PCR) were tested for their ability to detect marine birnavirus (MBV) in cultured rockfish, Sebastes schlegeli. One set of primers for RT-PCR was designed, based on a gene of infectious pancreatic necrosis virus (IPNV), and another set of primers for nested PCR was designed based on the VP2/NS junction region of MBV. This 2-step PCR method was specific for MBV and sensitivity was heightened when nested PCR was combined to RT-PCR. This 2-step PCR method was useful for detecting MBV not only in diseased fish, but also in asymptomatic fish. These results indicate that this 2-step PCR method is useful for detecting MBV in rockfish.

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Detection of RNA Mycoviruses in Wild Strains of Lentinula edodes in Korea

  • Kim, Eunjin;Park, Mi-Jeong;Jang, Yeongseon;Ryoo, Rhim;Ka, Kang-Hyeon
    • 한국균학회지
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    • 제49권3호
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    • pp.285-294
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    • 2021
  • In general, mycoviruses remain latent and rarely cause visible symptoms in fungal hosts; however, some viral infections have demonstrated abnormal mycelial growth and fruiting body development in commercial macrofungi, including Lentinula edodes. Compared to other cultivated mushrooms, L. edodes is more vulnerable to viral infections as it is still widely cultivated under near-natural conditions. In this study, we investigated whether Korean wild strains of L. edodes were infected by RNA mycoviruses that have previously been reported in other parts of the world (LeSV, LePV1, LeV-HKB, LeNSRV1, and LeNSRV2). Using specific primer sets that target the RNA-dependent RNA polymerase genes of each of the RNA mycovirus, reverse transcription-polymerase chain reaction (RT-PCR) was used to detect viral infection. Viral infection was detected in about 90% of the 112 wild strains that were collected in Korea between 1983 and 2020. Moreover, multiple infections with RNA mycoviruses were detected in strains that had normal fruiting bodies. This work contributes to our understanding of the distribution of RNA mycoviruses in Korea and the impact of multiple viral infections in a single strain of L. edodes.

Suitability of denervated muscle flaps as recipient sites for pancreatic islet cell transplantation

  • Park, Jong-Lim;Kim, Taewoon;Kim, Baek-Kyu
    • Archives of Plastic Surgery
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    • 제48권1호
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    • pp.133-143
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    • 2021
  • Background Extensive research has been conducted on islet transplantation as a possible cure for diabetes. Islet transplantation in the liver via the portal vein has shown remarkable results, but numerous other recipient sites are currently being investigated. We aimed to show the effectiveness of using a muscle flap as a recipient site for islet transplantation. Methods Islet cells were harvested from 12 isogenic Lewis rats, and then diabetes was induced in another 12 isogenic Lewis rats by streptozotocin injection. In six rats, 3,000 islets were transplanted into gastrocnemius muscle flaps, and in the other six rats, the same number of islets were transplanted into the gastrocnemius muscle. The transplanted islet cell function between the two groups was compared by means of blood glucose tests, glucose tolerance tests, immunohistochemistry, and real-time reverse transcription polymerase chain reaction. Results In the muscle flap group, blood glucose levels significantly decreased after islet transplantation. Blood glucose levels were significantly different between the two groups at 3 weeks after transplantation. The muscle flap group showed nearly normoglycemic results upon the glucose tolerance test, whereas the muscle group was hyperglycemic. Immunohistochemical evaluation showed positive results against insulin and glucagon in biopsies of both groups, and the islet cell density was higher in the muscle flap group. There were no statistically significant differences between the two groups in real-time reverse transcription polymerase chain reaction results. Conclusions Our results suggest that muscle flaps are promising candidates for islet cell transplantation.

Nucleic acid-based molecular diagnostic testing of SARS-CoV-2 using self-collected saliva specimens

  • Hwang, Eurim C.;Kim, Jeong Hee
    • International Journal of Oral Biology
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    • 제46권1호
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    • pp.1-6
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    • 2021
  • Since the outbreak of coronavirus disease 2019 (COVID-2019), the infection has spread worldwide due to the highly contagious nature of severe acute syndrome coronavirus (SARS-CoV-2). To manage SARS-CoV-2, the development of diagnostic assays that can quickly and accurately identify the disease in patients is necessary. Currently, nucleic acid-based testing and serology-based testing are two widely used approaches. Of these, nucleic acid-based testing with quantitative reverse transcription-PCR (RT-qPCR) using nasopharyngeal (NP) and/or oropharyngeal (OP) swabs is considered to be the gold standard. Recently, the use of saliva samples has been considered as an alternative method of sample collection. Compared to the NP and OP swab methods, saliva specimens have several advantages. Saliva specimens are easier to collect. Self-collection of saliva specimens can reduce the risk of infection to healthcare providers and reduce sample collection time and cost. Until recently, the sensitivity and accuracy of the data obtained using saliva specimens for SARS-CoV-2 detection was controversial. However, recent clinical research has found that sensitive and reliable data can be obtained from saliva specimens using RT-qPCR, with approximately 81% to 95% correspondence with the data obtained from NP and OP swabs. These data suggest that self-collected saliva is an alternative option for the diagnosis of COVID-19.

RT-RPA Assay Combined with a Lateral Flow Strip to Detect Soybean Mosaic Virus

  • Bong Geun Oh;Ju-Yeon Yoon;Ho-Jong Ju
    • The Plant Pathology Journal
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    • 제40권4호
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    • pp.337-345
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    • 2024
  • Soybean (Glycine max L.) is one of the most widely planted and used legumes in the world, being used for food, animal feed products, and industrial production. The soybean mosaic virus (SMV) is the most prevalent virus infecting soybean plants. This study developed a diagnostic method for the rapid and sensitive detection of SMV using a reverse transcription-recombinase polymerase amplification (RT-RPA) technique combined with a lateral flow strip (LFS). The RT-RPA and RT-RPA-LFS conditions to detect the SMV were optimized using the selected primer set that amplified part of the VPg protein gene. The optimized reaction temperature for the RT-RPA primer and RT-RPA-LFS primer used in this study was 38℃ for both, and the minimum reaction time was 10 min and 5 min, respectively. The RT-RPA-LFS was as sensitive as RT-PCR to detect SMV with 10 pg/µl of total RNA. The reliability of the developed RT-RPA-LFS assay was evaluated using leaves collected from soybean fields. The RT-RPA-LFS diagnostic method developed in this study will be useful as a diagnostic method that can quickly and precisely detect SMV in the epidemiological investigation of SMV, in the selection process of SMV-resistant varieties, on local farms with limited resources.