• 한국환경농학회지
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• 제26권4호
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• pp.319-324
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• 2007

오이모자이크바이러스(cucumber mosaic virus, CMV)에 저항성을 가지도록 개발된 형질전환 CMVP0-CP 고추를 이용하여 포장 조건에서 CMVP0-CP 고추 도입유전자의 지속성을 조사하였다. CMVP0-CP 고추의 잎을 토양 표면에 올려놓거나 토양 내에 묻은 뒤 일정 시간 간격으로 PCR 및 실시간 PCR법을 이용하여 도입유전자를 정성, 정량 분석하였다. CMVP0-CP 고추에 도입된 유전자의 양은 10 cm 깊이의 토양 속 및 토양 표면에서 1개월 후 28.3-42.7%, 2개월 후 0.9-3.3%로 감소하였으며, 3-4개월 후에는 검출되지 않았다. 또한 이러한 유전자는 지중보다 지표에서 약8% 더 오래 지속되었다. CMVP0-CP 고추의 잎이 토양 내에서 분해되는 과정에서 유출된 DNA가 주변 토양으로 전이되었는지를 조사하기 위해 낙엽주머니에 부착된 토양으로부터 DNA를 추출하여 PCR분석을 수행하였다. PCR 분석 결과 CMVP0-CP 고추에 도입된 유전자가 검출되지 않았다.

• Asian Pacific Journal of Cancer Prevention
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• 제13권3호
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• pp.1019-1024
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• 2012

Background: Infections with human papillomavirus (HPV) are highly prevalent among sexually active young women in India. However, not much is known about the incidence of type-specific human papillomavirus (HPV) infections and their patterns of persistence, especially in the Indian context. Objective: The objective of this study was to evaluate the rate of acquisition and persistence of HPV types in young women. Methods: Women residing in an urban slum in Delhi (n=1300) were followed for 24 months at 6 monthly intervals. Exfoliated cervical cells collected at each visit were tested for the presence of HPV DNA. Genotyping was performed using the reverse line blot assay. Results: The incidence rate for any HPV type was calculated to be 5 per 1000 women-months. Among high risk HPV types, HPV16 had the highest incidence rate followed by HPV59, HPV52 and HPV18, i.e., 3.0, 0.58, 0.41 and 0.35 women per 1000 women-months respectively. The persistence rate was higher for high-risk than low-risk HPV types. Among low-risk types, HPV42, HPV62, HPV84 and HPV89 were found to persist. Whereas almost all high risk types showed persistence, the highest rate was found in women with HPV types 16, 45, 67, 31, 51 and 59. The persistence rate for HPV16 infection was 45 per 1000 women-months. Conclusion: Incident HPV infections and high risk HPV type-specific persistence were found to be high in our study population of young married women. Understanding the patterns of HPV infection may help plan appropriate strategies for prevention programs including vaccination and screening.

• Molecules and Cells
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• 제45권1호
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• pp.33-40
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• 2022

The various DNA-protein interactions associated with the expression of genetic information involve double-stranded DNA (dsDNA) bending. Due to the importance of the formation of the dsDNA bending structure, dsDNA bending properties have long been investigated in the biophysics field. Conventionally, DNA bendability is characterized by innate averaging data from bulk experiments. The advent of single-molecule methods, such as atomic force microscopy, optical and magnetic tweezers, tethered particle motion, and single-molecule fluorescence resonance energy transfer measurement, has provided valuable tools to investigate not only the static structures but also the dynamic properties of bent dsDNA. Here, we reviewed the single-molecule methods that have been used for investigating dsDNA bendability and new findings related to dsDNA bending. Single-molecule approaches are promising tools for revealing the unknown properties of dsDNA related to its bending, particularly in cells.

• 환경생물
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• 제18권2호
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• pp.205-213
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• 2000

실험실에서 형질전환될 수 있는 세균들은 자연환경 조건에서도 형질전환 능력이 발달하는 것으로 알려져 있다. 따라서, 환경 내에서 형질전환 능력이 있는 세균의 존재는 확실한 것으로 여겨진다. DNA는 무기물에 부착된 상태에서는 핵산분해효소에 의한 분해로부터 보호되는 것으로 알려져 있다. 비록 DNA가 토양 속에 분산되어져 일정 비율로 가수분해되더라도, 수 주일 후에도 낮은 비율로 감지될 수 있다. 따라서 free DNA는 자연적 형질전환을 할 수 있을 만큼 충분히 지속될 수 있다. 실험실 조건에서는 세균의 형질전환이 여러 경우 보고되었지만, 자연상태에서 형질전환과 관련된 자료는 매우 적다. 생태학적으로 GMMs로부터 재조합 DNA가 토착 미생물에 전이될 수 있는 잠재력에 대한 문제가 주요 현안이 되었는데, 이는 전이된 DNA가 방출된 세균의 생태학적인 적응력을 변화시켜 생물학적 안전성의 문제를 야기할 수 있기 때문이다. 물론, 방출된 GMMs로부터 재조합 DNA가 토착 미생물에 전이되는 율은 아주 낮은 빈도로 일어나지만, 빈도가 낮다는 것은 그리 중요하지 않다. 왜냐하면, 비록 낮은 빈도로 전이되더라도 유리한 조건을 만나게 되면 전이된 유전자는 선택될 수 있기 때문이다. 이제까지 GMMs는 실험실이나 제한된 환경에서 주로 사용되었지만 앞으로는 개방된 자연 생태계에서 이루어질 전망이다. 그러므로 GMMs가 토착세균에 미치는 영향에 대해서도 연구되어야 하고 동시에 GMMs가 생태계에 방출될 경우 그에 따른 영향평가를 반드시 수행해야 한다.

• Journal of Microbiology and Biotechnology
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• 제25권8호
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• pp.1246-1256
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• 2015

Chlamydophila psittaci is an important intracellular pathogen. Persistent infection is an important state of the host-parasite interaction in this chlamydial infection, which plays a significant role in spreading the organism within animal populations and in causing chronic chlamydiosis and serious sequelae. In this study, a C. psittaci persistent infection cell model was induced by penicillin G, and real-time quantitative PCR was used to study the transcriptional levels of 10 C. psittaci genes (dnaA, dnaK, ftsW, ftsY, grpE, rpsD, incC, omcB, CPSIT_0846, and CPSIT_0042) in acute and penicillin-G-induced persistent infection cultures. Compared with the acute cultures, the penicillin-G-treated cultures showed a reduced chlamydial inclusion size and a significantly decreased number of elementary body particles. Additionally, some enlarged aberrant reticulate body particles were present in the penicillin-G-treated cultures but not the acute ones. The expression levels of genes encoding products for cell division (FtsW, FtsY) and outer membrane protein E encoding gene (CPSIT_0042) were downregulated (p < 0.05) from 6 h post-infection onward in the persistent infection cultures. Also from 6 h post-infection, the expression levels of DnaA, DnaK, IncC, RpsD, GrpE, and CPSIT_0846 were upregulated (p < 0.05); however, the expression level of OmcB in the persistent infection was< almost the same as that in the acute infection (p > 0.05). These results provide new insight regarding molecular activities that accompany persistence of C. psittaci, which may play important roles in the pathogenesis of C. psittaci infection.

• 한국토양비료학회지
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• 제48권5호
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• pp.442-450
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• 2015

This study was conducted to investigate the effect of Psy-2A-CrtI (PAC), a genetically modified (GM) rice with enhanced ${\beta}$-carotene, on the soil microbial community. The soil used to cultivate GM rice and its wild-type, Nakdong, was analyzed for population density, denaturing gradient gel electrophoresis (DGGE), and pyrosequencing. It was found that the bacterial, fungal and actinomycetes population densities of the PAC soils were within the range of those of the non-GM rice cultivar, Nakdong. The DGGE banding patterns of the GM and non-GM soils were also similar, suggesting that the bacterial community structures were stable within a given month and were unaffected by the presence of a GM plant. The pyrosequencing result showed a temporal difference in microorganism taxon and distribution ratio, but no significant difference between GM and non-GM was found. The persistence of the transgene DNA in the plant and surrounding soil were investigated for different time periods. There were differences in the persistence within the plant depending on the gene, but they could not be detected after 5 weeks. Also the transgenes were not detected in the surrounding soil. These results indicate that soil microbial communities are unaffected by the cultivation of a PAC rice within the experimental time frame.

• Asian Pacific Journal of Cancer Prevention
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• 제16권10호
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• pp.4297-4302
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• 2015

Background: To evaluate HPV testing by Hybrid Capture II (HCII) in conjunction with cytology in detecting the residual/recurrence disease after treatment of high-grade cervical intraepithelial neoplasia (CIN II-III) with loop electrosurgical excision procedure (LEEP). Materials and Methods: A retrospective review of 158 patients with histologically confirmed CIN II-III who underwent LEEP between January 2011 and October 2012 was conducted. Post-treatment control was scheduled at the 3rd, 6th, 12th and 18th month. All patients were followed up by Pap smear and HR-HPV genotype and viral load testing. Results: Pre-treatment, HR-HPV DNA, was detected in all specimens of the patients. At follow-up, 25 patients were diagnosed as the residual/recurrent disease during the FU visit, among whom, 16 patients with positive margin: 13 patients (52%) with HR-HPV DNA+/cytology+, 2 patients (8%) with HR-HPV DNA+/cytology-, 1 patient (4%) with cytology+/HR-HPV DNA-; 9 patients with clean margin - 5 patients (55.6%) with HR-HPV DNA+/cytology+; 2 patients (22.2%) with HRHPV DNA+/cytology-, 2 patients (22.2%) with cytology+/HR-HPV DNA-. None of them persisting HR-HPV DNA-/cytology-with positive or negative margin was identified as the residual/recurrent disease. The majority of residual/recurrent disease was detected at the 12th and 18th month FU, and there was almost no difference in the sensitivity and negative predictive value (NPV) between at the 3rd month and the 6th month FU visits. 14 residual/recurrence disease (14/46:30.4%) had pre-treatment high viral load (>5 000 RUL/PC) and 11 (11/112, 9.8%) with pre-treatment low viral load, P<0.05. Conclusions: (1) The persistence HR-HPV DNA is the root cause of the residual/recurrent disease for the women treated for high-grade CIN; the pre-treatment viral load and margin can be seen as the predictor. (2) The FU visit beginning at the 6th month post-treatment and lasting at least 24 months with the combination of cytology and HPV testing. (3) Patients with high pre-treatment HPV load, which is considered as one risk of developing the residual/recurrent disease, should be paid more attention (especially above 500RUL/PC) to by clinicians.

• 한국미생물학회:학술대회논문집
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• 한국미생물학회 2000년도 International Meeting 2000
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• pp.23-36
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• 2000

Japanese encephalitis virus (JEV) is the causative agent of a mosquito-borne encephalitis and is transmitted to human via persistently infected mosquito vectors. Although the virus is known to cause only acute infection, there were reports that showed neurological sequelae, latent infection in peripheral mononuclear cells, and recurrence of the disease after acute encephalitis. Innate resistance of certain cell lines, abnormal SN1 expression of the virus, and anti-apoptotic effect of cullular bcl-2 have been suggested as probable causes of JEV persistence even in the absence of defective interfering (DI) particles. Although possible involvement of DI particles in JEV persistence was suggested, neither has a direct evidence for DI presence nor its molecular characterization been made. Two questions asked in this study are whether the DI virus plays any role in JEV persistent infection if it is associated with and what type of change(s) can be made in persistently infected cells to avoid apoptosis even with the continuous virus replication, DI-free standard stock of JEV was infected in BHK-21, Vero, and SW13 cells and serial high multiplicity passages were performed in order to generate DI particles. There different-sized DI RNA species which were defective in both structural and nonstructural protein coding genes. Rescued ORFs of the DI genome maintained in-frame and the presence of replicative intermediate or replicative form RNA of the DI particles confirmed their replication competence. On the other hand, several clones with JEV persistent infection were established from the cells survived acute infections during the passages. Timing of the DI virus generation during the passages seemed coincide to the appearance of persistently infected cells. The DI RNAs were identified in most of persistently infected cells and were observed throughout the cell maintenance. One of the cloned cell line maintained the viral persistence without DI RNA coreplication. The cells with viral persistence released the reduced but continuous infectious JEV particle for up to 9 months and were refractory to homologous virus superinfection but not to heterologous challenges. Unlike the cells with acute infection these cells were devoid of characteristic DNA fragmentation and JEV-induced apoptosis with or without homologous superinfection. Therefore, the DI RNA generated during JEV undiluted serial passage on mammalian cells was shown to be biologically active and it seemed to be responsible, at least in part, for the establishment and maintenance of the JEV persistence in mammalian cells. Viral persistence without DI RNA coreplication, as in one of the cell clones, supports that JEV persistent infection could be maintained with or without the presence of DI particles. In addition, the fact that the cells with JEV persistence were resistant against homologous virus superinfection, but not against heterologous one, suggests that different viruses have their own and independent pathway for cytopathogenesis even if viral cytopathic effect could be converged to an apoptosis after all.

• 생명과학회지
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• 제13권1호
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• pp.99-104
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• 2003

Aromatic compounds are of major concern among environmental pollutants due to their toxicity and persistence. To monitor aromatic compounds in a real time with a better sensitivity, a new method of SPR (surface plasmon resonance) based on DNA chip (Biacore 3000) was developed here. It is thought that CapR regulatory protein as a complex with phenol, could bind to their corresponding promoter, Po. Biotinylated DNA oligomers for the promoter was synthesized by PCR and coupled onto streptoavidin-linked CM5-chip. CapR regulatory proteins were purified after cloning their genes in pET21a (+) vector and expressing proteins. The interaction was assessed by the system where the regulatory proteins flowed with or without phenol through the cells of DNA chip. CapR regulatory protein even in the presence of phenol had no response to its promoter, Po, suggesting that other factor(s) might be required for the activation of Po promoter. The present work reveals a promising possibility of the SPR-based DNA chip in monitoring specific environmental pollutants in a real time.

• Asian Pacific Journal of Cancer Prevention
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• 제15권19호
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• pp.8063-8067
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• 2014

Background: Cervical cancer is a major public health problem in Bangladesh. Persistence of high risk human papillomavirus (HRHPV) influences the progression of the disease, with an important role in followup for cervical intraepithelial neoplasia (CIN). Objective: To establish application of high risk HPV DNA test in the follow-up of women after treatment of CIN. Materials and Methods: This cross-sectional and hospital based study was carried out among 145 CIN treated women during the previous six months to three years at the colposcopy clinic of Bangabandhu Sheikh Mujib Medical University, Dhaka, between January 2011 and June 2012. Pap smear and HPV samples were collected and colposcopy was performed to find out the persistence of the disease. Cervical samples obtained were tested for HPV DNA using the Hybrid Capture II (HC-II) test. A cervical biopsy was collected whenever necessary. The results were compared to assess the efficacy of different methods during follow up such as Pap smear, HPV test and colposcopy. Results: Mean age of the recruited women (n=145) was 33.6 (${\pm}7.6$), mean age of marriage was 16.8 (${\pm}2.9$) and mean age of 1st delivery was 18.8 (${\pm}3.5$) years. More than half had high grade CIN before treatment and 115 (79.3%) women were managed by LEEP and 20.7% were managed by cold coagulation. Among the 145 treated women, 139 were negative for HPV DNA and six of them (4.1%) were HPV positive. Sensitivity of Pap smear (40.0) and HPV DNA test (40.0) was poor, but specificity was quite satisfactory (>93.0) for all the tests. Conclusions: The high risk HPV DNA test can be an effective method of identifying residual disease. It can be added to colposcopy and this should be applied to all treated women attending for their first or second post-treatment follow-up visit at 6 months to one year, irrespective of the grade of treated CIN.