• BMB Reports
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• 제39권1호
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• pp.26-36
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• 2006

Differential Display PCR technique (DD-PCR) was used for the analysis of altered gene expression in hemocytes of Vibrio harveyi-infected Penaeus monodon. Forty-four combinations of arbitrary and oligo(dT) primers were used to screen for differentially expressed genes. A total of 79 differentially expressed bands could be identified from 33 primer combinations. These included 48 bands (61%) whose expression level increased and 31 bands (39%) decreased after V. harveyi challenge. Subsequently, forty-eight differential display fragments were successfully reamplified and cloned. A total of 267 clones were randomly selected and sequenced. The sequence analysis showed that 85 (31%) out of 267 clones were matched with sequences in the GenBank database which represented 24 different genes with known functions. Among the known genes, glucose transporter 1, interferon-related developmental regulator 1, lysozyme, profilin, SERPINB3, were selected for further confirmation of their differentially expression patterns by real-time PCR. The results showed increasing in expression level of the selected genes in shrimp hemocytes after microbial challenge suggesting the involvement of such genes in bacterial response in shrimp. The anti-lipopolysaccharide factor type 3 (ALFPm3) gene, previously reported in P. monodon (Supungul et al., 2002) was found among the up-regulated genes but diversity due to amino acid changes was observed. Increase in ALFPm3 transcripts upon V. harveyi injection is in accordance with that found in the previous study.

• The Journal of Advanced Prosthodontics
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• 제10권4호
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• pp.308-314
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• 2018

PURPOSE. Plasma activation of hydrophobic zirconia surfaces might be suitable to improve the bond strength of luting materials. The aim of this study was to analyze the influence of nonthermal argon-plasma on the shear bond strength (SBS) between zirconia and different combinations of 10-MDP adhesive systems and luting composites after artificial aging. MATERIALS AND METHODS. Two hundred forty Y-TZP specimens were ground automatically with $165{\mu}m$ grit and water cooling. Half of the specimens received surface activation with nonthermal argon-plasma. The specimens were evenly distributed into three groups according to the adhesive systems ([Futurabond U, Futurabond M, Futurabond M + DCA], VOCO GmbH, Germany, Cuxhaven) and into further two subgroups according to the luting materials ([Bifix SE, Bifix QM], VOCO GmbH). Each specimen underwent artificial aging by thermocycling and water storage. SBS was measured in a universal testing machine. Statistical analysis was performed using ANOVA and $Scheff{\grave{e}}$ procedure with the level of significance set to 0.05. RESULTS. Surface activation with nonthermal plasma did not improve the bond strength between zirconia and the tested combinations of adhesive systems and luting materials. The plasma-activation trended to reveal higher bond strength if the self-etch luting material (Bifix SE) was used, irrespective of the adhesive system. CONCLUSION. Plasma-activation seems to be suitable to improve bond strength between zirconia and self-etch resin materials. However, further research is necessary to identify the influence of varying plasma-parameters.

• Journal of Microbiology and Biotechnology
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• 제27권7호
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• pp.1223-1232
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• 2017

Cordyceps militaris, a member of Ascomycota, a mushroom referred to as caterpillar Dong-chung-ha-cho, is commercially valuable because of its high content of bioactive substances, including cordycepin, and its potential for artificial cultivation. Cordycepin (3'-deoxyadenosine) is highly associated with the pharmacological effects of C. militaris. C. militaris is heterothallic in that two mating-type loci, idiomorph MAT1-1 and MAT1-2, exist discretely in two different spores. In this study, nine C. militaris strains were mated with each other to prepare newly bred strains that produced a larger amount of cordycepin than the parent strains. Nine strains of C. militaris were identified by comparing the internal transcribed spacer sequence, and a total of 12 single spores were isolated from the nine strains of C. militaris. After the MAT idiomorph was confirmed by PCR, 36 mating combinations were performed with six single spores with MAT1-1 and the others with MAT1-2. Eight mating combinations were successfully mated, producing stroma with perithecia. Cordycepin content analysis of all strains by high-performance liquid chromatography revealed that the KASP4-bred strain produced the maximum cordycepin among all strains, regardless of the medium and stroma parts. Finally, universal rice primer-PCR was performed to demonstrate that the bred strains were genetically different from the parental strains and new C. militaris strains. These results may be related to the recombination of genes during mating. The newly produced strains can be used to meet the industrial demand for cordycepin. In addition, breeding through mating suggests the possibility of producing numerous cordycepin-producing C. militaris strains.

• Parasites, Hosts and Diseases
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• 제34권3호
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• pp.191-196
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• 1996

중합반응(W_R)은 극미량의 핵산을 찾아내어 소수의 감염병원체를 확인하는 매우 민감한 진단법이다. 폐포자충 같이 다수의 숙주세포에 소수의 병원제가 섞여 있는 가검물에서 핵산의 정제 여부에 따른 중합반응의 민감도를 관찰하고. 특이도가 높은 시발제(primer)를 개발하기 위하여 이 연구를 수행하였다 흰쥐를 실험적으로 감염시키고 폐 폐포세척액. 혈청을 화보하여 현미경적 검사와 중합반응을 실시하였다 또한 사람과 횐쥐의 핵산을 위시하여 여러 미생물과 기생충. 이스트 의 핵산을 절제하여 이 시발체의 특이도를 검증하였다. 그 결과 여러 시발체 중에서 rRNA의 염기 서열 중에서 선택한 #24 주서열과 #27 대서열 쌍이 가장 우수한 민감도와 특이도를 보였다. 형태학적으로 양성인 폐포세척액의 세포용해액으로 반응시킨 경우 민감도가 57.7%이며 핵산을 정제한 경우 84.6%로 증가하였다 병원체 음성인 경우와 다른 병원체와 숙주의 핵산과는 반응하지 않았다. 혈청을 이용한 경우 20개 양성 표본 중 2개가 양성이고 6개의 감염된 흰쥐의 혈액은 모두 음성이었다. 충합반응을 폐포자충증의 진단에 활용하기 위하여는 폐포세척액 보다는 가래나 기관지 분비물. 혈청이나 혈액같은 비침습적인 가검물을 이용하고 핵산시료를 준비하는 과정이 간편하고 재현성이 있도록 개발되어야 할 것이다.

• Journal of Plant Biotechnology
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• 제44권3호
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• pp.303-311
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• 2017

본 연구는 참다래(Actinidia spp.) 유전자원의 유전적 다양성을 평가하기 위하여 재배품종 및 국내 외에서 수집한 참다래 61점을 대상으로 RAPD와 SRAP 분석을 수행하였다. RAPD 분석에서 40종의 선발 primer를 이용하여 230개의 다형성 밴드를 얻었으며, 평균 다형성 밴드 수는 5.75개였다. 32종의 primer 조합을 이용한 SRAP 분석에서 204개의 다형성 밴드를 획득하였고, 평균 다형성 밴드 수는 6.38 개였다. RAPD와 SRAP 분석에서 획득된 434개의 다형성 밴드를 이용하여 비가중 평균결합 방식으로 집괴분석한 결과 유전적 유사도 지수 0.680을 기준으로 3개의 그룹으로 분류되었다. 제1그룹에는 A. deliciosa와 A. chinensis에 속하는 품종과 A. deliciosa ${\times}$ A. arguta, A. chinensis ${\times}$ A. arguta, A. chinensis ${\times}$ A. deliciosa의 교잡종에 속하는 46점의 유전자원이 포함되었다. 제2그룹에는 A. arguta에 속하는 유전자원 7점과 A. arguta ${\times}$ A. deliciosa의 교잡종인 '스키니그린'이 포함되었다. 제3그룹에는 A. rufa, A. hemsleyana, A. macrosperma, A. polygama, A. eriantha 종에 속하는 유전자원 7점이 포함되었다. 참다래 유전자원 간 유전적 유사도 값은 0.479~0.991의 범위로 평균 유전적 유사도는 0.717이었다. 가장 높은 유사도 값(0.991)을 나타낸 유전자원은 NHK0038(A. deliciosa)과 NHK0040(A. deliciosa) 간이었고 가장 낮은 유사도 값(0.479)을 나타낸 유전자원은 '헤이워드'(A. deliciosa)와 K5-1-22(A. arguta) 간이었다.

• 한국자원식물학회지
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• 제17권3호
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• pp.227-238
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• 2004

본 연구는 자생식물 유전자원의 체계적 보존 및 관리를 위한 유전자원 수집과 보존 전략에 유용한 정보를 제공할 목적으로 강원 및 경북 지역에 자생하고 있는 꼬리 진달래 의 수집 계통들에 대하여 분자마커를 이용한 유전적 다형성 및 유연관계 분석을 수행하였다. RAPD분석에 이용된 10개의 primer에서 총 48개의 band가 관찰되었으며 이중에서 15개(31.3%)가 다형화 band였고, 1개의 primer당 평균 1.5개의 다형화 band가 관찰되었다. 반면에 AFLP 분석에서는 4개의 primer조합으로부터 총 62개의 band가 관찰되었으며 이중에서 33개(53.2%)가 다형화 band였고, 1개의 Primer조합당 평균 8.3개의 다형화 band가 관찰되었다. RAPD 및 AFLP band들을 이용한 UPGMA방법에 따라 작성된 dendrogram작성에서는 유전적 유사성 73.3%수준에서 크게 3개의 그룹으로 분리되었는데, 첫 번째 그룹에는 강원도 및 경북지역에서 수집된 대부분의 계통들이, 두 번째 그룹에는 lIII번 지역에서 수집된 7계통 모두가 포함되어 있었고 그리고 세 번째 그룹에는 경북 봉화에서 수집된 IV지역의 계통들 중 2계통(35, 36)이 각각 포함되었다. 따라서III번 지역과 IV번 지역의 일부 계통들을 제외하면 DNA 수준에서 꼬리 진달래 대부분의 계통들은 수집 지역에 따른 유전적 유연 관계를 명확히 나타내지 못하였다. RAPD및 AFLP분석 결과를 기초로 한 수집된 집단들 사이에서의 다형성 빈도는 II지역과 IV지역의 계통들에서 각각 45%로 가장 높게 관찰되었고, 반면에 Ⅵ지역의 계통들이 33%로 가장 낮게 관찰되었다. 그리고 집단들 사이에서의 유전적 유사성은 Ⅵ지역의 계통들이 평균 0.87로 가장 높게 나타났고, 반면에 I지역의 계통들은 평균 0.78을 나타내어 가장 낮게 나타났다. 본 연구의 결과에 의하면, 비록 III번 지역과 IV번 지역에서 수집된 일부 계통들은 다른 지역에서 수집된 대부분의 계통들과는 유전적으로 명확하게 구별되었지만, 수집 지역의 집단들 사이에서의 유전적 다양성은 현저한 차이를 나타내지 못하였으므로, 꼬리진달래의 현지외 보존을 통한 유전자원 보존원 조성을 효율적으로 수행하기 위해서는 유전자원 수집은 III번 지역과 IV번 지역을 중심으로 각 지역별로 균등하고 가급적 광범위하게 수집하는 것이 가장 효과적일 것으로 판단되었다.

• Mycobiology
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• 제31권2호
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• pp.68-73
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• 2003

Roots of Glycine max and Miscanthus sinensis and soil samples were collected from various field sites at Goesan, Chungbuk in Korea. Microscopic observations of the roots indicated high colonization rates of both arbuscular mycorrhizal fungi(AMF) and other fungi. The partial small subunit of ribosomal DNA genes were amplified with the genomic DNA extracted from their roots by nested polymerase chain reaction(PCR) with universal primer NS1 and fungal specific primers AML Restriction fragment length polymorphism(RFLP) was analyzed using the combinations of three restriction enzymes, HinfI, AluI and AsuC21. Nucleotides sequence analysis revealed that ten sequences from Miscanthus sinensis and one sequence from Glycine max were close to those of arbuscular mycorrhizal fungi. Also, 33% of total clones amplified with NS31-AM1 primers from M. sinensis and 97% from G. max were close to Fusarium oxysporum or other pathogenic fungi, and they were successfully distinguished from AME Results suggested that these techniques could help to distinguish arbuscular mycorrhizal fungi from root pathogenic fungi in the plant roots. Especially, DNA amplified by these primers showed distinct polymorphisms between AMF and plant pathogenic species of Fusarium when digested with AsuC21.

• Asian-Australasian Journal of Animal Sciences
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• 제16권2호
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• pp.277-281
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• 2003

Amplified fragment length polymorphism (AFLPs) markers were used to investigate the genetic variation in six autochthonous goat populations distributed in the middle and lower Yangtze River valley. The goat populations were Chengdu Grey Goat (CGG), Chuandong White Goat (CWG), Banjiao Goat (BG), Matou Goat (MG), Hui Goat (HG) and Yangtze River Delta White Goat (YRDWG). A total of 180 individuals (30 per population) were analysed using ten selected AFLP primer combinations that produced 78 clear polymorphism loci. The variability at AFLP loci was largely maintained within populations, as indicated by the average genetic similarity, and they were ranged from 0.745 to 0.758 within populations and 0.951 to 0.970 between populations. No breed specific markers were identified. Cluster analysis based on Nei' genetic distance between populations indicated that Chengdu Grey Goat is the most distant population, while CWG and YROWG were the closest populations, followed by BG, HG and MG. Genetic diversity of the goat populations didn' confirm what was expected on the basis of their geographical location, which may reflect undocumented migrations and gene flows and identify an original genetic resource.

• International Journal of Industrial Entomology and Biomaterials
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• 제17권2호
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• pp.211-215
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• 2008

To select entomopathogenic fungi controlling aphids effectively, several isolates were screened against second instars of Myzus persicae nymphs in the glasshouse using conidia suspension at $1.0{\times}10^5$ conidia/ml. Among these isolates, SFB-205 conidia showed the highest insecticidal activity about 32.7% efficacy to M. persicae at 4 days after application in the glasshouse. The attachment of SFB-205 conidia on the surface of M. persicae nymphs, and germination and penetration were observed using scanning electron microscopy. SFB-205 was identified as Beauveria bassiana species through the comparison of 5.8 s rRNA genes. There were 24 polymorphisms between SFB-205 and the previously reported isolate, B. bassiana ATCC74040 using six kinds of primer combinations in amplified fragment length polymorphism (AFLP) analysis. The B. bassiana SFB-205 might be used as a practical biological control agent for the green peach aphid, M. persicae in the field.

• Asian-Australasian Journal of Animal Sciences
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• 제19권8호
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• pp.1106-1110
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• 2006

In the meat industry, correct breed information in food labeling is required to assure meat quality. Genetic markers provide corroborating evidence to identify breed. This paper describes the development of DNA markers to discriminate between Japanese Black and F1 (Japanese Black${\times}$Holstein) breeds. The amplified fragment length polymorphism method was employed to detect candidate markers absent in Japanese Black but present in Holstein. The 1,754 primer combinations yielded eleven markers that were converted into single nucleotide polymorphism markers for high-throughput genotyping. The allele frequencies in both breeds were investigated for discrimination ability using PCR-RFLP. The probability of identifying F1 was 0.9168 and probability of misjudgment was 0.0066 using four selected markers. The markers could be useful for discriminating between Japanese Black and F1 and would contribute to the prevention of falsified breed labeling of meat.