• Mycobiology
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• 제30권4호
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• pp.202-207
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• 2002

This study was carried out to develop specific primers for PCR detection of Phellinus linteus. Diverse genomes of 15 Phellinus spp. including five Phellinus linteus isolates were fingerprinted by Primer Universal rice primer(URP)1F. The URP-PCR pattern differentiated P. linteus isolates from other phellinus spp. A polymorphic band(2.8 kb), which is unique for P. linteus isolates, was isolated and sequenced. Twenty four-oligonucleotide primer pairs were designed based on information of DNA sequence. The primer set(PLSPF2/PLSPR1) amplified single band(2.2 kb) of expected size with genomic DNA from seven Phellinus linteus, but not with that of other Phellinus species tested. The primers could be used identically in both DNA samples from mycelium and fruit bodies. This specific primers could offer a useful tool for detecting and identifying P. linteus rapidly.

• 한국양식학회지
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• 제19권2호
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• pp.67-76
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• 2006

Genomic DNA isolated from two Porphyra species, P. tenera and P. dentate from Wando located on the southern coast of Korean peninsula was amplified by PCR reaction. The amplified products were separated by agarose gel electrophoresis (AGE) with decamer primer and stained with ethidium bromide. The eight arbitrarily selected primers OPA-04, OPA-06, OPB-01, OPB-08, OPB-10, OPB-11, OPB-14 and OPC-10 generated the shared loci, polymorphic, and specific loci. The size of DNA bands varies from 100 bp to 2,200 bp. The complexity of the banding patterns varies dramatically between the primers and two Porphyra species. A total of 528 loci observed were identified in P. tenera and 443 in P. dentata: 22 polymorphic loci (4.2%) in P. tenera and 30 (6.8%) in P. dentata. 154 shared loci observed, the average 19.3 per primer, were identified in P. tenera and 143 loci, the aver-age 17.9 per primer, in P. dentata species. The number of specific loci in P. tenera and P. dentata was 73 and 77, respectively. The average bandsharing value was $0.623{\pm}0.008$ with P. tenera and $0.560{\pm}0.009$ within P. dentata. The average bandsharing value between two Porphyra species was $0.408{\pm}0.004$, ranged from 0.305 to 0.564. The dendrogram obtained by the eight primers indicates four genetic clusters. The genetic distance between two Porphyra species ranged from 0.076 to 0.627. The individual no. 02 of P. tenera was genetically closely related to no. 01 of P. tenera(genetic distance=0.082). Especially, two entities between the individual DENTATA no.21 and DENTATA no. 19 of P. dentata showed the longest genetic distance (0.627) in comparison with other individuals used. In this study, RAPD-PCR analysis has revealed the significant genetic distance between two Porphyra species pairs (P<0.001).

• Journal of Microbiology and Biotechnology
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• 제8권3호
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• pp.295-299
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• 1998

Oligonucleotide primers were designed and successfully applied to amplify DNA fragments of P450 hydroxylase genes from actinomycetes which produce a large variety of medically important metabolites. Primers were designed based on several regions of strong similarities in amino acid sequence of P450 hydroxylases from a variety of actinomycetes, primarily in the regions of an oxygen binding site and a heme ligand pocket. These primers were used to amplify DNA fragments from seven different actinomycetes species producing a variety of different compounds. The deduced amino acid sequences of the isolated fragments revealed significant similarities to known P450 hydroxylase including the product of the suaC or subC genes from Streptomyces griseolus that is capable of metabolizing a number of sulfonylurea herbicides, and to the product of the $P450_{sca2}$ from S. carbophilus that produces a specific HMG-CoA reductase inhibitor. This method should help researchers in cloning the P450 hydroxylase genes involved in the biosynthesis of useful compounds.

• Journal of Microbiology and Biotechnology
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• 제15권3호
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• pp.502-509
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• 2005

Sexual reproduction plays an important role in the biology and epidemiology of oomycete plant pathogens such as the heterothallic species Phytophthora infestans. Recent worldwide dispersal of A2 mating type strains of P. infestans resulted in increased virulence, gene transfer, and genetic variation, creating new challenges for disease management. To develop a genetic assay for mating type identification in P. infestans, we used the Amplified Fragment Length Polymorphism (AFLP) technique. The primer combination E+AT/M+CTA detected a fragment specific to A1 mating type (Mat-A1) of P. infestans. This fragment was cloned and sequenced, and a pair of primers (INF-1, INF-2) were designed and used to differentiate P. infestans Mat-A1 from Mat-A2 strains. The Mat A1-specific fragment was detected using Southern blot analysis of PCR products amplified with primers INF-1 and INF-2 from genomic DNA of 14 P. infestans Mat-A1 strains, but not 13 P. infestans Mat-A2 strains or 8 other isolates representing several Phytophthora spp. Southern blot analysis of genomic DNAs of P. infestans isolates revealed a 1.6 kb restriction enzyme (EcoRI, BamHI, AvaI)-fragment only in Mat-A1 strains. The A1 mating type-specific primers amplified a unique band under stringent annealing temperatures of $63^{\circ}C-64^{\circ}C$, suggesting that this PCR assay could be developed into a useful method for mating type determination of P. infestans in field material.

• 미생물학회지
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• 제39권3호
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• pp.166-174
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• 2003

Microcystin은 부영양화된 하천과 호수에서 cyanobacteria에 의해 생성되며 인간과 야생 동물에게 독성 물질로 작용하여 심각한 환경문제를 일으킨다. Microcystin은 mcy 유전자에 의해 암호화되는 microcystin synthetase로 알려진 multi-functional enzyme complex에 의해 리보좀의 관여 업이 합성된다. 따라서 mcy유전자의 PCR 증폭을 통해 독소를 생성하는 Microcystis를 효과적으로 검출할 수 있다. 본 연구에서는 microcystin을 생성하는 균주를 구별하기 위해 설계된 7종의 primer쌍의 유용성을 검증하기 위하여, 17주의 cyanobacteria와 국내 호수 시료에서 추출된 핵산을 대상으로 PCR 증폭을 하였다. TOX4E-TOX4R, FAA-RAA, FP-RP primer쌍에 의해서는 독소를 생성하는 Microcystis 균주 중 일부가 검출되지 않았다. NSZW2-NSZW1 primer쌍을 사용한 PCR의 경우 microcystin을 생성하는 국내 균주에서 예상하지 못한 크기의 산물이 관찰되었다. TOX1P-TOX1F primer쌍으로 PCR을 한 결과, 증폭된 산물을 관찰할 수 없었다. MSF-MSR과 TOX2P-TOX2F primer쌍만이 독소를 생성하는 11주의 Microcystis로부터 mcy유전자의 증폭을 성공하였다. 20개의 국내호수 시료에 각 Primer쌍에 의한 증폭여부를 확인한 결과, TOX2P-TOX2F primer쌍을 사용한 경우에만 모든 호수 시료에서 증폭된 산물을 관찰할 수 있었다. 본 연구 결과 TOX2P-TOX2F primer쌍이 국내 환경에서 독소를 생산하는 Microcystis를 검출하는데 가장 우수한 primer임을 확인할 수 있었다. 또한 Microcystis aenginosa NIER10010의 mcy 유전자의 염기서열 분석을 통해 국내 분리 균주의 유전적 다양성을 확인할 수 있었다.X> 을 일부 함유하고 있기 때문인 것으로 여겨진다. 궁극적으로 이 연구결과는 벤토나이트의 품위 산정에는 XRD 정량분석법이 적용되는 것이 합리적이고 CEC와 관련된 품질 특성은 전적으로 ‘Total CEC’ 개념에 의거하여 평가되어야 한다는 것을 시사한다.세균은 부유세균과는 다른 다양성을 이루고, 다른 천이과정을 거치는 것으로 확인되었다.로 interlayer된 것을 보여준다. 이와 같은 결과는 백운모, 녹니석 및 흑운모와 같은 변성 광물들이 비평형 광물 반응으로 만들어 졌으며 그 결과 불균질한 광물로 되었다는 것을 암시한다. led to the highest durability of all tested here. The reason of the improvement is due to thin MgF$_2$, which can prevent the $Mg_2$Ni electrode from forming Mg(OH)$_2$layer that is the main cause of degradation.platin에 의한 직접적 폐 독성은 발견되지 않았다이 낮았으나 통계학적 의의는 없었다[10.0%(4/40) : 8.2%(20/244), p>0.05]. 결론: 비디오흉강경술에서 재발을 낮추기 위해 수술시 폐야 전체를 관찰하여 존재하는 폐기포를 놓치지 않는 것이 중요하며, 폐기포를 확인하지 못한 경우와 이차성 자연기흉에 대해서는 흉막유착술에 더 세심한 주의가 필요하다는 것을 확인하였다. 비디오흉강경수술은 통증이 적고, 입원기간이 짧고, 사회로의 복귀가 빠르며, 고위험군에 적용할 수 있고, 무엇보다도 미용상의 이점이 크다는 면에서 자연기흉에 대해 유용한 치료방법임에는 틀림이 없으나 개흉술에 비해 재발율이 높고 비용이 비싸다는 문제가 제기되고 있는 만큼 더 세심한 주의와 장기 추적관찰이 필요하리라 사료된다.전 도부타민 심초음파는 관상동맥우회로술 후 동면심근의

• 한국균학회지
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• 제31권3호
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• pp.121-128
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• 2003

국내 유통균주에 대한 Phellinus속의 분류체계를 확립하고 종간구별을 위한 신속동정법을 개발하기 위해 계통학적 정보를 지니고 있는 ITS1, 5.8S rRNA및 ITS2부위의 염기서열을 밝히고 ITS1과 ITS2 부위의 다양한 염기서열을 이용하여 종 특이적인 유전자 탐침을 개발하였다. 본 연구에서 조사된 바에 의하면, 상황버섯으로 시판되고 있는 국내유통균주는 총 9균주로서 P. pini가 4종, P. baumii가 3종, P. igniarius와 P. linteus가 각각 1종으로 확인되었으며 P. gilvus은 조사되지 알았다. 그 중 P. pini는 대부분 중국에서 수입된 제품이었고 P. igniarius를 포함한 P. baumii는 주로 북한산 제품으로 조사되었다. 한편 P. linteus와 P. baumii는 변이가 높은 ITS1 , ITS2 부위에서도 다른 종에 비하여 높은 상동성을 나타내어 종 특이적인 유전자 탐침이 유효하지 않은 반면에 P. igniarius, P. pini, P. gilvus종에 대한 detection primer로 각각 IF1-IR3, PF1-PF3, GF2-FR4 primer는 유전자 탐침이 유효함을 확인하였다. 유연관계가 아주 가까운 P. linteus와 P. baumii에 대한 정확한 분석을 위해서는 변이가 심한 ITS 부위와 보존성이 높은 18S rDNA 부위의 염기서열을 함께 분석, 비교하여야 가능하리라 사료된다.

• 한국약용작물학회지
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• 제20권4호
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• pp.277-285
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• 2012

In this study, Expressed Sequence Tag-Simple Sequence Repeat (EST-SSR) analyses were used to clarify the genetic polymorphisms among Korean ginseng cultivars and breeding lines and to classify them into distinct genetic groups. Polymorphic and reproducible bands were produced by 14 primers out of total 30 primers used in this study. Fourteen EST-SSR loci generated a total of 123 bands. Amplified PCR products showed the highly reproducible banding patterns at 110~920 bp. The number of amplified bands for each EST-SSR primers ranged from 2 to 19 with a mean of 8.8 bands. P26 and P35 primers showed 13 and 12 banding patterns, respectively. The number of alleles for each EST-SSR locus ranged from 1.67 to 2.00 with a mean of 1.878 alleles. P34 and P60 primers showed the highest and the lowest genetic polymorphism, respectively. Cluster analysis based on genetic similarity estimated by EST-SSR markers classified Korean cultivars and breeding lines into 4 groups. Group included Gopoong and Chunpoong and 9 breeding lines (55%), group included 2 breeding lines (10%), group included 3 breeding lines (15%), group included Gumpoong and 3 breeding lines (20%). Consequently, the EST-SSR marker developed in this study may prove useful for the evaluation of genetic diversity and differentiation of Korean ginseng cultivars and breeding lines.

• 대한치과보철학회지
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• 제39권4호
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• pp.417-432
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• 2001

The pcf metal primers on the bond strength and durability of 4-META/MMA-TBB resins adhered to an Au-Ag-Pd alloy. For this study, the specimens were divided into 8 groups as follows: Thermocyle 0 : (1) control group : sandblast, (2) Group I : sandblast + Cesead Opaque Primer; (3) Group II : sandblast + Metal Primer; (4) Group III : sandblast + V-Primer; Thermocyle 10,000 (5) control sandblast: (6) Group I : sandblast + Cesead Opaque Primer: (7) Group II : sandblast + Metal Primer; (8) Group III sandblast + V-Primer. The shear bond strength was determined using an Instron were observed with the use of scanning electron microscope. Finally, the strengths of bonded joints were evaluated with regard to their adherence energy using a wedge test. The results obtained were as follows ; (1) The shear bond strength of 4-META/MMA-TBB resin to the Au-Ag-Pd alloy was significantly improved in all the groups treated with the primers (p<0.05). (2) Regardless of the adhesive primers used, a significant difference was observed in the bond strength of the thermocycle 0 groups and 10,000 groups (p<0.05). (3) Both before and after thermocycling, the strongest bond strength between the resin an the alloy was obtained after treatment with a metal primer containing MEPS (p<0.05). (4) In the wedge test, the adherence energies of the control group and Group III decreased more rapidly than those of Group I and II during the 2nd day of storage in water.

• Mycobiology
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• 제42권1호
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• pp.46-51
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• 2014

A degenerated strain of Pleurotus eryngii KNR2312 was isolated from a commercial farm. Random amplified polymorphic DNA analysis performed on the genomic DNA of the normal and degenerated strains of this species revealed differences in the DNA banding pattern. A unique DNA fragment (1.7 kbp), which appeared only in the degenerated strain, was isolated and sequenced. Comparing this sequence with the KNR2312 genomic sequence showed that the sequence of the degenerated strain comprised three DNA regions that originated from nine distinct scaffolds of the genomic sequence, suggesting that chromosome-level changes had occurred in the degenerated strain. Using the unique sequence, three sets of PCR primers were designed that targeted the full length, the 5' half, and the 3' half of the DNA. The primer sets P2-1 and P2-2 yielded 1.76 and 0.97 kbp PCR products, respectively, only in the case of the degenerated strain, whereas P2-3 generated a 0.8 kbp product in both the normal and the degenerated strains because its target region was intact in the normal strain as well. In the case of the P2-1 and P2-2 sets, the priming regions of the forward and reverse primers were located at distinct genomic scaffolds in the normal strain. These two primer sets specifically detected the degenerate strain of KNR2312 isolated from various mushrooms including 10 different strains of P. eryngii, four strains of P. ostreatus, and 11 other wild mushrooms.

• Journal of Animal Science and Technology
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• 제47권4호
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• pp.615-624
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• 2005

본 연구는 배양초기 pH 조건이 F. succino- genes의 섬유소 부착과 섬유소 소화에 미치는 영향을 보고자 실시하였다. 선정된 specific primer를 이용하여 F. succinogenes의 genomic DNA로부터 445bp의 16S rDNA 절편을 증폭하여 205bp의 internal control을 제작하였고, cPCR 결과로부터 박테리아 수를 계산할 수 있는 표준곡선의 회귀식($r^2$>0.99)을 얻을 수 있었다. In vitro 배양초기 pH에 따른 F. succ- inogenes의 cellulose 부착을 cPCR로 모니터링한 결과, 발효과정 전 기간동안 초기 pH가 6.8과 6.2일 때 cellulose 건물 g당 부착 균주의 수가 pH 5.6일 때 보다 높았으나, pH 6.8과 6.2 사이에서는 큰 차이는 없었다(p>0.05). Cellulose 분해는 배양시간이 진행됨에 따라 증가 되었으며, 분해 정도는 pH가 증가함에 따라 더 높았다. 배양초기 pH가 6.8, 6.2 그리고 5.8일 때 48시간동안 감소한 pH는 각각 0.24, 0.58 그리고 0.16 이었다. 가스 생산량은 발효 시간이 경과함에 따라 pH가 높을수록 더 많았다. 결론적으로 발효 초기 pH는 F. succinogenes에 의한 cellulose 소화, 가스 생산, pH 변화 및 cellulose 부착에 큰 영향을 주었으며, 특히, 낮은 pH(5.8)에서는 섬유소 소화 및 박테리아 부착을 현저한 감소 시켰다.