• 분석과학
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• 제12권1호
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• pp.80-83
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• 1999

사람에게서 유래된 DNA시료의 X,Y 특이의 alphoid gene을 PCR법으로 분석하면 성별을 확인할 수 가 있다. PCR법으로 alphoid gene을 분석한바 매우 예민도가 높아 genomic DNA 약 60pg까지 성별을 분석할 수 있었다. 그리고 성별이 혼합되어 있는 DNA에서 female DNA의 1/10비 까지는 male DNA를 분석할 수 있었다. 따라서 이 결과는 혼합된 DNA에서 X,Y 특이의 alphoid gene을 분석하는데 기준으로 활용할 수가 있다.

• 한국육종학회지
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• 제43권5호
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• pp.448-456
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• 2011

적색 과육 '홍양' 품종에서 차등발현하는 유전자를 찾기 위하여 mirror orientation selection (MOS)과 결합된 suppression subtractive hybridization (SSH) 실험을 수행하였다. 그 결과, 288개의 cDNA clone을 확보하였으며, colony PCR을 통해 192개의 positive clone을 선발하였고, 이들을 sequencing하였다. NCBI/Genbank 데이터베이스의 BLAST 검색를 이용하여 염기서열을 분석한 결과, 30개의 clone에서 기존에 알려진 유전자기능과의 유사성을 확인할 수 있었으며, 10개의 clone이 특이유전자였다. 그 중 3개의 clone(AcF21, AcF42, AcF106)은 과실 후숙과 관련된 ACC-oxidase와 상동성이 있었다. SSH의 결과를 통해 얻어진 이 유전자들의 차등발현양상을 확인하기 위하여 reverse transcription PCR(RT-PCR)과 quantitative real-time PCR(qReal-time PCR) 분석을 실시하였다. qReal-time PCR분석과 RT-PCR분석에서 모두 동일한 결과를 확인할 수 있었으며, 3개 clone 모두 '홍양'에서의 유전자발현수준이 '헤이워드'보다 더 높았다. AcF21은 다른 유전자들보다 가장 높은 발현수준을 나타내었는데, 만개 후 120일과 160일 모두 '홍양'에서의 발현수준이 높았다.

• Fisheries and Aquatic Sciences
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• 제5권1호
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• pp.54-58
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• 2002

The polymerase chain reaction (PCR) amplification technique was used for comparison of Listeria monocytogenes serotypes. PCR primers for the fragment of invasion-associated protein (iap) gene were highly specific for all the serotypes of L. monocytogenes. Other Listeria spp., such as Listeria ivanovii and Listeria innocua were not produced the PCR fragments by above primer set. The nucleotide sequences of PCR products showed high homologies in comparison of all the isolated serotypes except unknown type II-2. The deduced amino acid sequences of the PCR products also showed similar to one another. The various region of the PCR products, called a Thr-Asn repeat region was presented. All of isolated L. monocytogenes serotypes possessed 16 to 20 Thr-Asn repeats.

• 한국식물병리학회지
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• 제14권4호
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• pp.308-313
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• 1998

Reverse transcription and polymerase chain reaction (RT-PCR) techiniques were used to identification and differentiation of cucumber mosaic virus isolated from Forsythia koreana (CMV-Fk). RT-PCT used by two set of 20-mer primers one was CMV-common primers and another was CMV subgroup I-specific primers designed in a conserved region of the 3' end of CMV RNA3, amplified about 490 bp and 200 bp DNA fragments from CMV-Fk, respectively. CMV could be detected by RT-PCR at a dilution as low as 10-4 in forsythia crude sap extracts. Restriction enzyme analysis of RT-PCR products using EcoRI and MspI showed that CMV-Fk belonged to CMV subgroup I. But, analysis of RNA fingerprinting by arbitrarily primed polymerase chain reaction (RAP-PCR) showed heterogeneity of RNA3 between CMV-Fk and CMV-Y as a member of subgroup I.

• 한국치위생학회지
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• 제17권2호
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• pp.319-330
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• 2017

Objectives: The purpose of this study was to compare SYBR Green qPCR, TaqMan, and bacterial selective medium cultures for accurate quantitative analysis of oral microorganisms. Methods: The SYBR Green method is widely used to analyze the total amount of oral microorganisms in oral saliva. However, in this study, MTR-PCR method based on TaqMan method was performed using newly developed primers and probes. In addition, it was designed to confirm the detection agreement of bacteria among bacteria detection method. Results: As a result of MRT-PCR and SYBR Green qPCR analysis, more than 40 times (0.9-362.9 times) bacterium was detected by MRT-PCR. In addition, more bacteria were detected in saliva in the order of MRT-PCR, SYBR Green qPCR, and bacterium culture, and the results of MRB-PCR and SYBR Green qPCR showed the highest agreement. The agreement between the three methods for detecting P. intermedia was similar between 71.4 and 88.6%, but the agreement between MRT-PCR and SYBR Green qPCR was 80% for S. mutans. Among them, the number of total bacteria, P. intermedia and S. mutans bacteria in saliva was higher than that of SYBR Green qPCR method, and bacterium culture method by MRT-PCR method. P. intermedia and S. mutans in saliva were detected by MRT-PCR and MRT-PCR in 88.6% of cases, followed by the SYBR Green qPCR method (80.0%). Conclusions: The SYBR Green qPCR method is the same molecular biology method, but it can not analyze the germs at the same time. Bacterial culturing takes a lot of time if there is no selective culture medium. Therefore, the MRT-PCR method using newly developed primers and probes is considered to be the best method.

• 미생물학회지
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• 제37권1호
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• pp.56-60
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• 2001

탄저균의 분자적 다양성 분석은 다양한 DNA표지의 부족으로 쉬운 일이 아니어서, 본 연구에서는 random amplified polymorphic DNA (RAPD)-PCR을 이용하여 Bacillus 속으로부터 탄저균을 구별할 수 있는 새로운 DNA 표지를 개발하고자 하였다. RAPD-PCR을 이용한 분석은 다양한 Bacillus 종으로부터 탄저균을 동정할 수 있었으며, 아울러 Bacillus 종 사이에서 확실한 유전적인 변이를 확인할 수 있었다. 이러한 분석은 간단, 신속하고, 그리고 정확하게 탄저균을 진단하는데 활용할 수 있다고 본다.

• 식물병연구
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• 제17권2호
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• pp.169-176
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• 2011

우리나라에서 분리한 P. katsurae의 유전적 특성을 구명하기 위하여 국내에서 분리한 P. katsurae를 대상으로 nuclear DNA(nDNA)의 ${\beta}$-tubulin (BTU)과 Elongation facter 1 alpha (EF1A) 그리고 rDNA ITS 부위의 PCR-SSCP 분석을 실시하여, P. katsurae와 Phytophthora 속에 속하는 다양한 종들의 각 부위를 대상으로 유전적 유연관계를 비교분석 하고 동정에 이용하고자 하였다. 각각의 Phytophthora 속에서 변이가 가장 많이 발생하는 부위를 포함하여 증폭 시킬 수 있도록 각 부위의 공통 염기배열로부터 제작된 primer는 Phytophthora 종에 특이적인 반응을 나타냄으로서 동정 및 진단에도 유용하게 활용될 수 있을 것으로 판단되었다. SSCP 분석 결과는 국내 P. katsurae 균주와 공시한 다른 Phytophthora 속 균주들과의 구분이 가능하였으며, Phytophthora 종 간의 구분도 가능하였다. 그러나 한 가지 부위만을 이용한 PCR-SSCP 분석은 Phytophthora 종 간의 구분이 어려운 경우도 있었다. 따라서 보다 정확하고 명확한 Phytophthora 종의 유전적 다양성 분석 및 동정을 위하여서는 단일 부위에 의한 PCR-SSCP보다는 복수 부위에 의한 PCR-SSCP를 실시하는 것이 바람직한 것으로 확인되었다.

• Fisheries and Aquatic Sciences
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• 제7권3호
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• pp.130-135
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• 2004

The marine dinoflagellate genus Alexandrium, which produces PSP toxins, has a global distribution. As human-assisted dispersal of the species has been suggested, it is important to develop molecular tools to trace the dispersal pathway. To screen population-specific DNA sequences that differentiate Korean and Japanese A. catenella, we targeted the area downstream of the chloroplast psbA gene using PCR with population-specific DNA primers followed by RFLP (restriction fragment length polymorphism) analysis and sequencing. The RFLP patterns of the PCR products divided Korean and Japanese A. catenella regional isolates into three types: Korean, Japanese, and type CMC3, isolated from Korea. We sequenced the PCR products, but found no similar gene in a homology search. The molecular phylogeny inferred from the sequences separated the Korean and Japanese A. catenella strains, as did the RFLP patterns. However, the Japanese isolates included two slightly different sequences (types J and K), while the Korean sequence was the same as the Japanese K type. In addition, a unique sequence was found in the Korean strains CMC2 and CMC3. Population-specific PCR amplification with Japanese A. catenella type-specific PCR primers designed from the type J sequence yielded PCR products for Japanese strains only, showing that the unknown gene can be used for a population analysis of Korean and Japanese A. catenella.

• 대한임상검사과학회지
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• 제41권4호
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• pp.153-157
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• 2009

The production of extended-spectrum ${\beta}$-lactamases ($ESBL_S$) of the TEM or SHV type by bacterial pathogens is a major threat to the use of the clinically important expanded-spectrum cephalosporins. The characterization of the SHV ESBLs producing Klebsiella pneumoniae strains present in clinical isolates is time-consuming processes. We describe here in the development of a novel system, which consists of a real time PCR. We found 11 K. pneumoniae strains to be presumptive strains ESBLs producers by clinical and laboratory standards institute (CLSI) guidelines. The double disk synergy test showed 8 ESBL positive and conventional PCR showed 10 SHV ESBL positive, which were K. pneumoniae strains isolates. By real time PCR analysis, SHV gene in 11 of 11 strains were identified. When sequencing analysis was compared with real time PCR, both analysis were presented 99% similarity. In this study, we used a rapid, sensitive, and specific real-time PCR (RT-PCR) method for detection of the assay SHV ESBL producing K. pneumoniae strains in clinical isolates.

• 생태와환경
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• 제37권1호통권106호
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• pp.122-129
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• 2004

In an effort to develop the biomarker for monitoring the contamination of xenoestrogen in the freshwater environment of Korea, reverse transcription-polymerasechain reaction (RT-PCR) analysis of vitellogenin (VTG) gene expression was optimized in Hearisarsus Iaseo, Based on the homology of the VTG cDNA sequences between the common carp and zebra fish, a set of PCR primers for VTG mRNA amplification for H; labo was designed. VTG mRNA level in livers from female and male fishes was analyzed by RT-PCR following single injection of 17 beta estradiol($E_2$ 10 mg $kg^{-1}$ B.W.). As an internal control, beta actin mRNA was amplified. One us of total liver RNA was subjected to RT-PCR. In female the amount of PCR productof VfC gradually increased in the range from 16 to 34 cycles of amplification. On the contrary, in control male, PCR product first detected at 32 cycles of amplification and linearly increased up to 40 cycles of amplification. In $E_2$ injected male liver, the VTC mRNA level was similar to that in the female. Taken together, this result suggests that liver of male H. labo expresses minute amount of VTG mRNA which are2-l6 equivalent of female and that induction of VTG mRNA occurs in male liver after estrogen treatment. In conclusion, the optimized protocol for RT-PCR analysis of VTG mRNA expression in liver of male H. labo will provide the environmental monitoring method for the xenoestrogen contamination in the rivers in Korea.