• 환경생물
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• 제17권3호
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• pp.279-284
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• 1999

토양으로부터 직접 추출한 DNA를 cross hybridization하는 방법을 통해서 서로 다른 토양 환경 간에 미생물 군집의 유전형적 유사성과 상대적 다양성을 비교하였다. 그 결과 소나무삼림토양이 다른 토양에 비해 상대적 다양성이 높은 것으로 밝혀졌으며, 경작지, 나지, 초지, 신갈삼림 순으로 다양성 정도를 나타내었다. 또한 유전형적 유사성의 정도에 따른 집괴 분석 결과 소나무삼림과 경작지 토양, 신갈나무삼림과 초지 토양 그리고 나지 등 세 부류로 구분되었다.

• BMB Reports
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• 제37권4호
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• pp.439-444
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• 2004

Double stranded targets on the cDNA microarray contain representatives of both the coding and noncoding strands, which will introduce hybridization competition with probes. Here, the effect of double and single strands of targets on the signal intensity and the ratios of Cy5/Cy3 within the same slide were compared. The results show that single stranded targets can increase the hybridization efficiency without changing the Cy5/Cy3 ratio. Based on these results, a new strategy was established by generating cDNA targets with asymmetric PCR, instead of conventional PCR, to increase the sensitivity of the cDNA microarray. Furthermore, the feasibility of this approach was validated. The results indicate that the cDNA microarray system based on asymmetric PCR is more sensitive, with no decrease in the reliability and reproducibility as compared with that based on conventional symmetric PCR.

• 한국미생물·생명공학회지
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• 제14권2호
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• pp.133-137
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• 1986

원생동물인 Tetrahymena thermophila의 17S-rDNA구조 및 hygromycin 내성 기구에 대한 연구의 일부로서 hygromycin 내성변이주 hmr3의 17S-rDNA를 대장균의 vector pBR 322에 cloning하였다. 우선 rDNA는 hot phenol-cresol 용액으로 추출하여 제한효소 Hind III 처리로서 약 2.2kbp의 17S-rDNA를 agarose 전기영동상에서 분리하였다. 이를 pBR 322에 cloning하여 wild type의 17S-rDNA probe와 colony hybridization시켜 선별하였다. 그중 5-19 균주의 recombinant plasmid로부터 17S-rDNA 의 전사 orientation위치가 pBR322의 tetracyline내성 유전자 쪽으로 삽입되어 있는 것을 확인하였다.

• The Plant Pathology Journal
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• 제37권3호
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• pp.291-298
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• 2021

False smut caused by Ustilaginoidea virens is an important rice fungal disease that significantly decreases its production. In the recent past, conventional methods have been developed for its detection that is time-consuming and need high-cost equipments. The research and development in nanotechnology have made it possible to assemble efficient recognition interfaces in biosensors. In this study, we present a simple, sensitive, and selective oxidized graphene-based geno-biosensor for the detection of rice false smut. The biosensor has been developed using a probe DNA as a biological recognition element on paper electrodes, and oxidized graphene to enhance the limit of detection and sensitivity of the sensor. Probe single-stranded DNA (ssDNA) and target ssDNA hybridization on the interface surface has been quantitatively measured with the electrochemical analysis tools namely, cyclic voltammetry, and linear sweep voltammetry. To confirm the selectivity of the device, probe hybridization with non-complementary ssDNA target has been studied. In our study, the developed sensor was able to detect up to 10 fM of target ssDNA. The paper electrodes were employed to produce an effective and cost-effective platform for the immobilization of the DNA and can be extended to design low-cost biosensors for the detection of the other plant pathogens.

• 한국수의병리학회지
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• 제7권1호
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• pp.1-4
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• 2003

Malignant catarrhal fever (MCF) is a systemic disease of ruminants caused by a gamma herpesvirus, ovine herpesvirus 2 (OvHV-2). Dot blot hybridization (DBH) protocols for detecting and differentiating this MCF virus were developed. OvHV-2 specific primer pairs, 556/555, were used for the amplification of target DNA. Then, the amplified DNA was labeled with incorporation of digoxigenin (DIG). The Dig-labeled probe was able to detect and differentiate specifically OvHV-2 DNA. This DBH technique can be applied to confirm the presence of MCF virus on clinical samples and to differentiate specifically between OvHV-2 infection and other viral infections.

• 대한의생명과학회지
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• 제21권1호
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• pp.32-39
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• 2015

High-risk (HR) human papillomavirus (HPV) genotypes are strongly associated with cervical cancer, whereas other HPV genotypes are not. To identify the various HPV genotypes in clinical samples, we conducted HPV genotyping using a DNA chip test and reverse blot hybridization assay (REBA) in normal cytology samples and atypical squamous cells of undetermined significance (ASCUS) cytology samples. We also investigated the HPV infection rate and HPV genotype prevalence in women with normal cytology and ASCUS cytology. Liquid-based cytology preparations were used for the initial screening of 205 subjects with normal cytology and ASCUS cytology. The HPV infection rate was 49.8% when using the DNA chip assay and 61.0% when using the REBA test. In patients with normal cytology, the HR-HPV positive rate was 21.9% with the DNA chip assay and 43.9% with the REBA test. In contrast, 8.3% of patients with ASCUS were HR-HPV positive when using the DNA chip assay, and 13.6% were positive when tested with the REBA test. The infection rate of HR-HPV in the 40~50-year age group was significantly higher than that of the other age groups. Based on the cytological analysis of the normal and ASCUS samples, the five most prominent HPV genotypes were HPV 16, 18, 68, 33, and 58 using the DNA chip test, and they were HPV 16, 18, 53, 33, and 66 when using the REBA test. In conclusion, the findings show that the results of the REBA test are comparable to those of the DNA chip test. Most strikingly, the REBA test detected the HR-HPV genotype associated with cervical carcinoma similar to that detected with the DNA chip method. Therefore, the REBA test is a useful method to detect clinically important HR-HPV genotypes.

• 생명과학회지
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• 제13권4호
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• pp.522-528
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• 2003

본 연구는 출아형 효모 Saccharomyces cerevisiae에서 자외선의 상해 시 이를 정상으로 회복시키는 절제회복 (excision repair) 유전자로 알려진 RADA4의 특성 규명을 위하여 균류 Coprinus cinereus에서 이와 유사한 유전자를 분리하였다. RAD4 유사 유전자를 분리하기 위하여 균류 C. cinereus의 염색체 DNA를 전기영동하여 분리한 다음 효모 RAD4 DNA를 probe로하여 이와 hybridization하였다. 이 결과 RAD4 유사 유전자는 3.2 kb의 insert DNA를 갖고 있었다. 또한 Southern hybridization으로 이 유사 유전자는 fungus C. cinereus의 염색체에 존재함을 확인하였다. 분리한 RAD4 유사 유전자의 전사체 크기는 2.5 kb 였으며, 자외선의 상해 시 전혀 'inducibility가 없음을 Northern hybridization으로 확인하였다. 또한 유사유전자 부분을 삭제하였을 때 이 부분이 없는 세포는 전혀 생존을 못하였다. 이 결과 분리한 RAD4 유사유전자는 세포의 생존에 관여함을 알 수 있었다.

• 한국발생생물학회지:발생과생식
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• 제21권3호
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• pp.237-247
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• 2017

Mammalian spermatogenesis occurs in a precise and coordinated manner in the seminiferous tubules. One of the attempts to understand the detailed biological process during mammalian spermatogenesis at the molecular level has been to identify the testis specific genes followed by study of the testicular expression pattern of the genes. From the subtracted cDNA library of rat testis prepared using representational difference analysis (RDA) method, a complimentary DNA clone encoding type III member of a DnaJ family protein, DnaJC18, was cloned (GenBank Accession No. DQ158861). The full-length DnaJC18 cDNA has the longest open reading frame of 357 amino acids. Tissue and developmental Northern blot analysis revealed that the DnaJC18 gene was expressed specifically in testis and began to express from postnatal week 4 testis, respectively. In situ hybridization studies showed that DnaJC18 mRNA was expressed only during the maturation stages of late pachytene, round and elongated spermatids of adult rat testis. Western blot analysis with DnaJC18 antibody revealed that 41.2 kDa DnaJC18 protein was detected only in adult testis. Immunohistochemistry study further confirmed that DnaJC18 protein, was expressed in developing germ cells and the result was in concert with the in situ hybridization result. Confocal microscopy with GFP tagged DnaJC18 protein revealed that it was localized in the cytoplasm of cells. Taken together, these results suggested that testis specific DnaJC18, a member of the type III DnaJ protein family, might play a role during germ cell maturation in adult rat testis.

• 대한전기학회:학술대회논문집
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• 대한전기학회 2006년도 제37회 하계학술대회 논문집 C
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• pp.1322-1323
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• 2006

This research aims to develop the multiple channel electrochemical DNA chip that has the above characteristic and be able to solve the problems. At first, we fabricated a high integration type DNA chip array by lithography technology. It is able to detect a plural genes electrochemically after immobilization of a plural probe DNA and hybridization of non-labeling target DNA on the electrodes simultaneously.

• 한국미생물·생명공학회지
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• 제34권2호
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• pp.101-108
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• 2006

Phytophthora sp.의 균사성장을 특이적으로 저해하는 토양 미생물인 YNB54 균주의 정확한 분류적 위치를 밝히기 위해 Biolog GN2, API 20E와 같은 상업적 생화학 kit, 16S rDNA, DAN-DNA hybridization, GC함량, MIDI 등의 분석을 수행하였다. 다양한 생화학적 kit를 사용한 동정 결과는 이 균주가 다른 어떤 종보다 Enterobacter cloacae와 E. cancerogenus에 보다 더 가까움을 보여주었다. 또한 DAN-DNA hybridization, GC함량, MIDI 분석의 결과들 역시 다른 속 (Citerobacter, Klebsiella, Leclercia)보다 Enterobacter 속에 더 유사함을 암시해 주었다. 그러나 16S rDNA분석에서 이 균주는 Citrobacter freundii(99.4%)와 동일 그룹으로 구분되었지만 Enterobacter, Leclecia, Klebsiella 속 등과도 98%이상의 상동성을 보여주는 polyphyletic 특성을 보였다. 결론적으로 YNB54의 분류 동정을 위한 우리의 조사들은 이 균주가 유전적으로 다양하고 지금까지 아는 것보다 분류학적으로 더 복잡함을 암시해줌에도 불구하고 Enterobacter속임이 가장 유력하다는 것을 보여 주었다.