• 아시안잔디학회지
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• 제23권2호
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• pp.307-316
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• 2009

크리핑벤트그래스는 골프코스의 퍼팅그린에서 주로 사용하고 있는 한지형잔디 초종이다. 크리핑벤트그래스 품종들은 형태적인 특성이나 유전적인 다양성이 협소하여 품종간 구분이 매우 어렵다. 따라서 이들 품종간 유전적인 다양성이나 골프코스 그린의 인터씨딩율에 대한 조사를 위해 SCAR marker를 개발하고자 본 연구를 수행하였다. penncross, penn A-4, crenshaw, L-93, CY-2, T-1 공시품종들에 대한 SCAR marker 개발을 위해 5개 RAPD primer에 대한 품종 간 구분이 가능한 특이적 band를 선발하였다. 선발한 특이적 band의 염기서열을 분석하여 품종별 SCAR primer를 제작하였다. 각 품종의 SCAR primer별로 특이적 band가 나타나는지에 대한 여부를 검정한 결과, penncross, penn A-4, crenshaw 품종의 SCAR primer는 6개 공시품종에서 동일한 크기의 band가 나타나 SCAR marker로써 활용이 어려웠다. 하지만 CY-2의 CY850F/R primer는 850bp, T-1의 T700F/R primer는 700bp, L-93의 L2900F/R는 2.9kb에서 특이적 band가 나타나 3개 품종별 SCAR marker로서 활용이 가능하였다. 따라서 본 연구에서 개발한 3개 품종별 SCAR marker를 활용하여 크리핑벤트그래스의 품종별 유전적 다양 및 골프코스 그린의 인터씨딩율 조사에 활용이 가능할 것으로 기대된다.

• 한국동물위생학회지
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• 제35권4호
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• pp.289-294
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• 2012

Mycoplasmas are highly fastidious bacteria, difficult to culture and slow growing. Infections with Mycoplasma species can cause a variety of problems in living organisms and in vitro cell cultures. In this study, we investigated the usefulness of a genus-specific consensus PCR analysis method to detect Mycoplasma species. The developed consensus primer pairs MycoF and MycoR were designed specifically to amplify the 16S ribosomal RNA gene (rRNA) of Mycoplasma species by the optimized PCR system. The developed consensus PCR system effectively amplified 215 bp of Mycoplasma genus-specific region of 16S rRNA. In conclusion, we recommend this consensus PCR for monitoring Mycoplasma species in animals, human and cell culture system.

• Journal of Microbiology and Biotechnology
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• 제17권5호
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• pp.806-811
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• 2007

Bacillus anthracis is a soil pathogen capable of causing anthrax that is closely related to several environmental species, including B. cereus, B. mycoides, and B. thuringiensis. DNA homology studies showed that B. anthracis, B. cereus, B. mycoides, and B. thuringiensis are closely related, with a high sequence homology. To establish a method to specifically detect B. anthracis in situations such as environmental contamination, we initially performed RAPD-PCR with a 10-mer random primer and confirmed the presence of specific PCR bands only in B. anthracis species. One region specific for B. anthracis was cloned and sequenced, and an internal primer set was designed to amplify a 241-bp DNA fragment within the sequenced region. The PCR system involving these specific primer sets has practical applications. Using lyses methods to prepare the samples for PCR, it was possible to quickly amplify the 241-bp DNA segment from samples containing only a few bacteria. Thus, the PCR detection method developed in this study is expected to facilitate the monitoring of environmental B. anthracis contamination.

• 산업기술연구
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• 제28권A호
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• pp.141-147
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• 2008

Several contaminated bacteria such as Lactobacillus brevis and Pediococcus damnosus in beer production cause beer spoilage by producing off flavours and turbidity. Detection of these organisms is complicated by the strict anaerobic conditions and lengthy incubation times required for their cultivation, consequently there is a need for more rapid detection methods. Recently, two contaminated strains were isolated from vessel of beer production and identified as Lactobacillus species by API kit identificaton as well as 16S-23S ITS sequencing analyses. Two isolated strains were named as Lactobacillus sp. HLA1 and Lactobacillus HLB2, respectively. A polymerase chain reaction (PCR) method was developed for the rapid and specific detection of Lactobacillus sp.. Two sets of primer pairs (HLA1-F/HLA1-R and HLB2-F/HLB2-R) were designed for the amplification of a 1576 base pair (bp) fragment of the HLA1 16S-23S rRNA gene and 1888 bp fragement of the HLB2 16S-23S rRNA. Amplified PCR products were highly specific to detect corresponding bacteria when other contaminated strains were used as PCR templates. However, detection of both strains were limited when $100{\mu}{\ell}$ of cultured samples were mixed with $100m{\ell}$ of beer sample in arbitrary manner. The sensitivity of the assay still needs to be improved for direct detection of the small amounts of bacteria present in beer.

• The Plant Pathology Journal
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• 제15권5호
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• pp.287-294
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• 1999

A technique based on the polymerase chain reaction (PCR) for the specific detection of genus Phytophthora and Phytophthora cryptogea-P. drechsleri complex group was developed using nucleotide sequence information of ribosomal DNA (rDNA) regions. The internal transcribed spacers (ITS) including 5.8S were sequenced for P. cryptogea-P. drechsleri complex group and its related species. Two pairs of oligonucleotide primers were designed. Primer pair ITS1/Phy amplified ca. 240 bp fragment in 12 out of 13 specie of Phytophthora, but not in Pythium spp., Fusarium spp.and Rhizoctonia solani. Primer pair rPhy/Pcd amplified 549 bp fragment only in P. cryptogea-P. drechsleri complex group, but not in other Phytophthora spp.and other genera. Specific PCR amplification using the primers was successful in detecting Phytophthora and P. cryptogea-P. drechsleri complex group in diseased plants.

• International Journal of Oral Biology
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• 제38권2호
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• pp.43-49
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• 2013

The objective of this study was to develop PCR primers that are specific for Streptococcus sanguinis, Streptococcus parasanguinis, and Streptococcus gordonii. We designed the S. sanguinis-, S. parasanguinis-, and S. gordonii-specific primers, Ssa21-F3/Ssa21-R2, Spa17-F/Spa17-R, and Sgo41-F1/Sgo41-R1 respectively, based on the nucleotide sequences of the Ssa21, Spa17, and Sgo41 DNA probes that were screened using inverted dot blot hybridization (IDBH). The species-specificity of these primers was assessed against 43 strains of mitis group streptococci, including clinical strains of S. sanguinis, S. parasanguinis, and S. gordonii. The resulting PCR data revealed that species-specific amplicons had been obtained from all strains of the target species tested, and that none of these amplicons occurred in any other strains from other species. These results suggest that the Ssa21-F3/Ssa21-R2, Spa17-F/Spa17-R, and Sgo41-F1/Sgo41-R1 primers may be useful in detecting S. sanguinis, S. parasanguinis, and S. gordonii at the species level, respectively.

• 식물병연구
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• 제18권2호
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• pp.73-79
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• 2012

Phytophthora katsurae는 밤나무 잉크병의 원인이 되는 병원성 균류이다. P. katsurae를 검출하기 위하여 2개의 duplex PCR primer set을 제작하였다(SOPC 1F/1R+KatI 3F/5R, SOPC 1-1F/1-1R+KatI 3F/5R). SOPC 1F/1R과 SOPC 1-1F/1-1R primer pair는 sequence characteristic amplification regions(SCAR)를 이용하여 제작하였고, KatI 3F/5R primer pair는 internal transcribed spacer(ITS) 부위로부터 제작하였다. 추출한 genomic DNA를 1 mg/ml에서 1 ng/ml까지 10배씩 순차적으로 희석하여 균사량에 따른 Duplex PCR의 민감도를 확인한 결과, 2개의 primer set은 각각 1 ${\mu}g/ml$와 100 ng/ml까지 검출이 가능하였다. 유주포자를 $1{\times}10^6$부터 $1{\times}10^2$ cells/ml까지 10배씩 순차적으로 희석하고 genomic DNA를 추출하여 P. katsurae의 유주포자량에 의한 검출 한계를 확인한 결과, 2개의 primer set 모두 $1{\times}10^5$ cells/ml까지 검출할 수 있었다. 각각의 primer set은 PCR 검출을 실시하였을 때 P. katsurae 균주에서만 PCR 산물이 증폭된 반면에, Phytophthora 16종에서는 P. katsurae에 특이적인 PCR 증폭산물이 생성되지 않았다. 따라서, 2개의 primer set을 이용한 Duplex PCR은 P. katsurae의 신속하고 효과적인 검출에 유용하게 활용될 수 있을 것이다.

• Fisheries and Aquatic Sciences
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• 제6권1호
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• pp.13-19
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• 2003

Genomic DNA from the muscle of marsh clam (Corbicula leana) from Gochang, Muan and a Chinese site was extracted to identify genetic differences and similarity by randomly amplified polymorphic DNAs-polymerase chain reaction (RAPD- PCR). Out of 20 primers, seven primers produced amplified fragments which were consistently polymorphic. A total of 1,246 amplified products were produced of which 530 were polymorphic $(42.5\%)$. The number of polymorphic bands produced per primer varied from 40 to 122 with an average of 75.7 in marsh clam from Gochang. 3.28 of the 23.0 polymorphic bands per lane were found to be polymorphic. Also, about $4.34\%$ of total polymorphic bands were specific to marsh clam from Gochang. The major common bands of 0.28 kb generated by primer OPB-15 (GGAGGGTGTT) were present in every individuals, which were polymorphic. This common bands in every individuals should be diagnostic of specific strains, species and/or their relatedness. Primer OPB-19 (ACCCCCGAAG) produced the highest number of 12 specific bands. The intra-population variation was revealed in the band patterns identified by this primer. The random primer OPB-12 (CCTTGACGCA) yielded the amplified fragments which were consistently polymorphic between the marsh clams from Gochang and from Muan. This primer produced a total of 77 polymorphic bands: 31 bands from Gochang, 14 from Muan and 32 from the Chinese populations. An average of polymorphic bands were 1.8 from Gochang and 2.5 from the Chinese populations. This value obtained from the Chinese population was higher than those from the two domestic populations. Generally, the RAPD polymorphism generated by these primers may be useful as a genetic marker for strain or population identification of marsh clam.

• 생명과학회지
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• 제25권8호
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• pp.903-909
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• 2015

LAMP (Loop-mediated Isothermal Amplification)법은 PCR를 기반으로 등온에서 autocycling 가닥 변위 DNA 합성에 의존하며, Bst polymerase를 사용하여 진단하는 방법이다. 이것은 대상 DNA의 여섯 개의 배열을 인식하는 4개의 특정 primer의 도움을 받아 단시간 안에 병원체를 식별하는 높은 특이성을 지니고 있다. 본 연구에서는 LAMP로 수생에서 위험한 병원체인 Vibrio alginolyticus의 특별한 LAMP primer를 제작하였으며, 신속한 진단을 위해 MgSO₄, dNTP, Betaine, Bst polymerase의 최적 반응 조건의 특이성 및 기존의 PCR보다 10배 정도의 민감하다는 것을 확인하였다. 또한, 디자인 되어진 LAMP primer가 다른 Vibrio 종들 중 오직 V. alginolyticus에서만 반응한 것을 확인 할 수 있었다. 본 논문에서는 병원체 세균인 V. alginolyticus의 빠르고 민감한 효과적인 진단으로 양식 질병들을 조기에 발견할 수 있도록 개발하였다.

• Journal of Microbiology and Biotechnology
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• 제28권12호
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• pp.1992-1998
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• 2018

In 2015, Bacillus paralicheniformis was separated from B. licheniformis on the basis of phylogenomic and phylogenetic studies, and urease activity was reported as a phenotypic property that differentiates between the two species. Subsequently, we have found that the urease activity of B. paralicheniformis is strain-specific, and does not reliably discriminate between species, as strains having the same urease gene cluster were identified in B. licheniformis and B. sonorensis, the closest relatives of B. paralicheniformis. We developed a multilocus sequence typing scheme using eight housekeeping genes, adk, ccpA, glpF, gmk, ilvD, pur, spo0A, and tpi to clearly identify B. paralicheniformis from closely related Bacillus species and to find a molecular marker for the rapid identification of B. paralicheniformis. The scheme differentiated 33 B. paralicheniformis strains from 90 strains formerly identified as B. licheniformis. Among the eight housekeeping genes, spo0A possesses appropriate polymorphic sites for the design of a B. paralichenofomis-specific PCR primer set. The primer set designed in this study perfectly separated B. paralicheniformis from B. licheniformis and B. sonorensis.