• 한국산학기술학회논문지
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• 제6권2호
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• pp.183-188
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• 2005

포유동물에서는 두가지의 receptor가 보고되었다. 각 sample을 RNA extraction, RT-PCR, Realtime-PCR을 실시하여 melatonin 1A, 1B의 발현 양을 분석하였다. MT1A는 cerebellum에서는 3 hours에서 약 1/8배로 감소를 보이고 6 hours에서는 정상치 9 hours에서는 16배정도로 많은 양의 증가율 보였다. 나머지 hippocampus, thalamus, hypothalamus에서는 공통적으로 3 hours에서 많게는 10배에서, 적게는 대조군과 거의 비슷한 1.5배정도의 증가율을 보이고 있으며, 6 hours에서는 모두 감소하는 것을 알 수 있다. MT1B에서는 4 group 모두 3 hours, 6 hours에서 receptor의 양이 확연히 줄어들었다. 9 hours의 경우에는 4 group모두에서 적게는 8배, 많게는 거의 1000배 가까이의 발현 양의 차이가 나타났다.

• 대한약침학회지
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• 제15권2호
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• pp.20-23
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• 2012

Panax ginseng is a well-known herbal medicine in traditional Asian medicine, and wild ginseng is widely accepted to be more active than cultivated ginseng in chemoprevention. However, little has actually been reported on the difference between wild ginseng and cultivated ginseng. Thus, to identify and analyze those differences, we used suppressive subtraction hybridization (SSH) sequences with microarrays, realtime polymerase chain reaction (PCR), and reverse transcription PCRs (RT-PCRs). One of the clones isolated in this research was the chloroplast rpoC1 gene, a ${\beta}$subunit of RNA polymerase. Real-time RT-PCR results showed that the expression of the rpoC1 gene was significantly upregulated in wild ginseng as compared to cultivated ginseng, so, we conclude that the rpoC1 gene may be one of the important markers of wild ginseng.

• The Plant Pathology Journal
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• 제35권2호
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• pp.164-171
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• 2019

An assay for detecting Apple scar skin viroid (ASSVd) was developed based on nucleic acid sequence based amplification (NASBA) in combination with realtime detection during the amplification process using molecular beacon. The ASSVd specific primers for amplification of the viroid RNA and molecular beacon for detecting the viroid were designed based on highly conserved regions of several ASSVd sequences including Korean isolate. The assay had a detection range of $1{\times}10^4$ to $1{\times}10^{12}$ ASSVd RNA $copies/{\mu}l$ with reproducibility and precision. Following the construction of standard curves based on time to positive (TTP) value for the serial dilutions ranging from $1{\times}10^7$ to $1{\times}10^{12}$ copies of the recombinant plasmid, a standard regression line was constructed by plotting the TTP values versus the logarithm of the starting ASSVd RNA copy number of 10-fold dilutions each. Compared to the established RT-PCR methods, our method was more sensitive for detecting ASSVd. The real-time quantitative NASBA method will be fast, sensitive, and reliable for routine diagnosis and selection of viroid-free stock materials. Furthermore, real-time quantitative NASBA may be especially useful for detecting low levels in apple trees with early viroid-infection stage and for monitoring the influence on tree growth.

• Restorative Dentistry and Endodontics
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• 제33권6호
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• pp.537-544
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• 2008

세균의 검출에 있어서 polymerase chain reaction (PCR) 방법은 기존의 plate counting과 달리 빠르게 세균을 검출할 수 있다. 하지만 세균이 죽은 후에도 DNA는 장기간 존재할 수 있기 때문에, DNA에 기초한 분석은 살아있는 세균과 죽은 세균을 구분할 수 없다. 최근에 DNA extraction전에 propidium monoazide (PMA)를 처리하여 살아있는 세균만 선택적으로 검출하는 방법이 제시되었다. PMA는 손상된 세포막만 통과하여 죽은 세포의 DNA와 빛 노출 하에서 결합하여 PCR이 증폭되는 것을 막는다. Enterococcus faecalis는 근관치료의 실패에 있어서 중요한 원인이 되는 세균으로 제시되어 왔다. 그리고 chlorhexidine (CHX)은 E. faecalis의 제거에 있어서 효과적인 약제임이 밝혀졌다. 이번 실험의 목적은 세균 수의 측정에 있어서, PMA 처리와 real-time PCR 방법의 적용 가능성을 기존의 plate counting과 비교하여 알아보는 것이다. 또한 E. faecalis에 대한 2% CHX의 살균 효과를 PMA 처리 후 real-time PCR 방법을 사용하여 알아보는 것이다. 실험 방법으로 먼저 살아있는 세균과 죽은 세균을 다른 비율로 섞어서 PMA를 처리한 후 real-time PCR을 시행하여 PMA가 빛 노출 하에서 죽은 세균의 DNA와 결합하는 효과를 나타내는지 알아보았다. 다음으로 PMA 처리 후 realtime PCR 방법을 이용하여 살아있는 세균의 양을 측정한 것을 plate counting으로 얻은 CFU와 비교하였다. 마지막으로 2% CHX의 처리시간을 다르게 하였을 때 E. faecalis에 대한 살균 효과를 PMA 처리 후 real-time PCR 방법을 사용하여 알아보았다. 실험 결과로 살아있는 E. faecalis의 비율이 감소할수록 Ct value는 증가하였다. 그리고 PMA 처리 후 real-time PCR 방법을 이용하여 세균의 양을 측정한 것과 plate counting으로 얻은 CFU 사이에는 Optical density (OD) 값이 1.0일 때까지는 상관관계가 있었다. 하지만 OD 값이 1.5일 때는, PMA를 처리한 후 real-time PCR을 시행했을 때 측정된 살아있는 세균의 양이 감소하였음에 반해서 plate counting에 의한 CFU는 계속 증가하였다. 마지막으로 2% CHX을 오래 적용할수록 살아있는 E. faecalis의 상대적인 양이 감소하는 것을 PMA 처리와 real-time PCR 방법을 이용해 확인하였다.

• 한국수정란이식학회지
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• 제26권4호
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• pp.237-244
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• 2011

Ski protein is implicated in proliferation/differentiation in a variety of cells. We had previously reported that Ski protein is present in granulosa cells of atretic follicles, but not in preovulatory follicles, suggesting that Ski has a role in apoptosis of granulosa cells. The alternative fate of granulosa cells other than apoptosis is to differentiate to luteal cells, however, it is unknown whether Ski is expressed and has a role in granulosa cells undergoing luteinization. Thus, the aim of the present study was to examine whether the initiation of luteinization with luteinizing hormone (LH) directly regulates expression of Ski in the luteinized granulosa and luteal cells after ovulation by in vitro models. RT-PCR and real time PCR analysis respectively revealed that LH had no effect on c-Ski mRNA expression in the cultured granulosa cells regardless of LH treatment. Though Ski protein is absent in granulosa cells of preovulatory follicle, its mRNA (c-Ski) was expressed and the level was unchanged even after LH surge. Taken together, these results demonstrated that Ski protein expression is induced in granulosa cells upon luteinization, and suggested that its expression is regulated post-transcriptionally. Moreover, expression of mRNA of Arkadia, an E3 ubiquitin ligases, in luteinizing granulosa cells in vivo was assessed by realtime-PCR. The levels of Arkadia mRNA expression were unchanged during follicular growth and postovulatory luteinization. These findings suggest that Ski protein level may be regulated during luteinization at translational and/or post-translational level but not by Arkadia.

• The Plant Pathology Journal
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• 제34권4호
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• pp.286-296
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• 2018

Maintenance of a beneficial microbial community, especially in the rhizosphere, is indispensable for plant growth and agricultural sustainability. In this sense, plant growth-promoting rhizobacteria (PGPR) have been extensively studied for their role in plant growth promotion and disease resistance. However, the impact of introducing PGPR strains into rhizosphere microbial communities is still underexplored. We previously found that the Proteus vulgaris JBLS202 strain (JBLS202) promoted growth of Kimchi cabbage and altered the relative abundance of total bacteria and Pseudomonas spp. in the treated rhizosphere. To extend these findings, we used pyrosequencing to analyze the changes in bacterial communities in the rhizosphere of Kimchi cabbage after introduction of JBLS202. The alterations were also evaluated by taxon-specific realtime PCR (qPCR). The pyrosequencing data revealed an increase in total bacteria abundance, including specific groups such as Proteobacteria, Acidobacteria, and Actinobacteria, in the treated rhizosphere. Time-course qPCR analysis confirmed the increase in the abundance of Acidobacteria, Actinobacteria, Alphaproteobacteria, and Betaproteobacteria. Furthermore, genes involved in nitrogen cycling were upregulated by JBLS202 treatment indicating changes in ecological function of the rhizosphere soil. Overall, these results indicate that introduction of JBLS202 alters both the composition and function of the rhizosphere bacterial community, which can have direct and indirect effects on plant growth. Therefore, we propose that long-term changes in bacterial composition and community-level function need to be considered for practical use of PGPRs.

• Journal of Microbiology and Biotechnology
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• 제23권7호
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• pp.993-1003
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• 2013

Methanotrophs are the most important sink of $CH_4$, which is a more highly potent greenhouse gas than $CO_2$. Methanotrophic abundance and community diversity in cover soils from two typical semi-aerobic landfills (SALs) in China were detected using real-time polymerase chain reaction (real-time-PCR) and denaturing gradient gel electrophoresis (DGGE) based on 16S rRNA genes, respectively. Real time-PCR showed that Type I methanotrophs ranged from $1.07{\times}10^6$ to $2.34{\times}10^7$ copies/g soil and that of Type II methanotrophs from $1.51{\times}10^7$ to $1.83{\times}10^8$ copies/g soil. The ratio of Type II to Type I methanotrophic copy numbers ranged from 5.61 to 21.89, indicating that Type II methanotrophs dominated in SAL. DGGE revealed that Type I methanotrophs responded more sensitively to the environment, changing as the community structure varied with different soil types and locations. Methylobacter, Methylosarcina, and Methylomicrobium for Type I, and Methylocystis for Type II were most prevalent in the SAL cover layer. Abundant interflow $O_2$ with high $CH_4$ concentration in SALs is the reason for the higher population density of methanotrophs and the higher enrichment of Type II methanotrophs compared with anaerobic landfills and other ecosystems, which proved a conclusion that increasing the oxygen supply in a landfill cover layer would greatly improve $CH_4$ mitigation.

• 대한치과의료관리학회지
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• 제6권1호
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• pp.11-18
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• 2018

The aim of this study was to evaluate the competency of the Easyperio test, a genetic test method based on real time PCR for the detection of bacteria that cause dental caries and periodontal disease. To verify the validity of this text, various dental health evaluations were administered to 33 boys between the ages of 12 to 14, as this age group commonly experiences dental caries. These evaluations included a dental caries experience survey, a first molar health evaluation, the Dentocult Streptococcus mutans (SM) strip mutans, the Dentocult Lactobacillus spp (LB) test, and the Easyperio test. The correlation coefficients between the level of the Dentocult SM strip mutans and the dental caries experience were DT (R=0.570, p=0.001), DMFT (R=0.376, p=0.031), and first molar health (R=-0.395, p=0.023). The correlation coefficients between the amount of SM in the Easyperio test and dental caries experience were DT (R=0.528, p=0.002), DMFT (R=0.369, p=0.035), and first molar health (R=-0.426, p=0.013). The correlation coefficients between the level of the dentocult SM strip mutans and the SM amounts of the Easyperio test were S.mi (R=0.564 p=0.001) and S.mu (R=0.621, p=0.002). The correlation coefficients between the level of the Dentocult LB test and the SM amount of Easyperio test was S.mi (R=0.495, p=0.003). In conclusion, Easyperio test may be an easy and effective method for the differentiation and diagnosis of dental caries through quantitative and qualitative analysis of oral bacteria.

• 생약학회지
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• 제52권4호
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• pp.228-233
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• 2021

This research was carried out to investigate the moisturizing effects of Agarum cribrosum extract on normal human epidermal keratinocytes (NHEKs). Moisturizing effects of A. cribrosum extract on NHEKs were measured by quantitative realtime RT-PCR to verify the gene expressions related to skin hydration, hyaluronic acid (HA)-ELISA to detect HA production, and cell viability assays. A. cribrosum extract increased the mRNA levels of the AQP3 and HAS2 genes and HA production in NHEKs. On the other hand, A. cribrosum extract decreased the mRNA level of the KRT1 and KRT10 genes known as differentiated keratinocyte marker in NHEKs. This research showed the moisturizing effects of A. cribrosum extract. The results indicate that A. cribrosum extract can be a potent functional ingredient for skin hydration and anti-aging products. Further study is warranted regarding the use of A. cribrosum extract to develop not only cosmetics but also food and medicine.

• Clinical and Experimental Pediatrics
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• 제54권10호
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• pp.405-408
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• 2011

Purpose: In autumn 2009, the swine-origin influenza A (H1N1) virus spread throughout South Korea. The aims of this study were to determine the clinical characteristics of children infected by the 2009 H1N1 influenza A virus, and to compare the rapid antigen and realtime polymerase chain reaction (PCR) tests. Methods: We conducted a retrospective review of patients ${\geq}18$ years of age who presented to Soonchunhyang University Hospital in Seoul with respiratory symptoms, including fever, between September 2009 and January 2010. A real-time PCR test was used to definitively diagnose 2009 H1N1 influenza A infection. Medical records of confirmed cases were reviewed for sex, age, and the time of infection. The decision to perform rapid antigen testing was not influenced by clinical conditions, but by individual factors such as economic conditions. Its sensitivity and specificity were evaluated compared to real-time PCR test results. Results: In total, 934 patients tested positive for H1N1 by real-time PCR. The highest number of patients (48.9%) was diagnosed in November. Most patients (48.2%) were aged between 6 and 10 years. Compared with the H1N1 real-time PCR test results, the rapid antigen test showed 22% sensitivity and 83% specificity. Seventy-eight patients were hospitalized for H1N1 influenza A virus infection, and fever was the most common symptom (97.4%). Conclusion: For diagnosis of 2009 H1N1 influenza A virus infection, the rapid antigen test was inferior to the real-time PCR test in both sensitivity and specificity. This outcome suggests that the rapid antigen test is inappropriate for screening.