• Asian-Australasian Journal of Animal Sciences
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• 제19권11호
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• pp.1519-1523
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• 2006

Through a screening and selection approach method, decamer random markers were used in a technique called random amplified polymorphic DNA (RAPD) assay with 252 genomic DNAs isolated from four major Haimen chicken populations: Rugao (62), Jiangchun (62), Wan-Nan (63) and Cshiqishi (65). A total of 3-score decamer random primers (S241-S260, S1081-S1100 and S1341-S1360) were employed in the preliminary RAPD-polymerase chain reaction (RAPD-PCR) assay with 50 random template DNA samples from all the populations. Four (6.67%) of the primers that produced obvious polymorphic patterns, interpretable and reproducible bands were selected and used with both the individual DNAs from each population and with pooled DNA samples of the four populations in subsequent analyses. The selected primers produced a total of 131 fragments with molecular size ranging from 835 to 4,972 base pairs (bp) when used with the individual DNAs; 105 (80.15%) of these fragments were polymorphic. With the pooled DNAs, 47 stable and characteristic bands with molecular size ranging from 840 to 4,983 bp, of which 23 (48.94%) polymorphic, were also generated. The band-sharing coefficient (BSC) calculated for the individuals in the population and among populations of bulked samples was between 0.8247 (Rugao) and 0.9500 (Cshiqishi); for pairwise populations, it was between 0.7273 (Rugao vs. Wan-Nan) and 0.9367 (Jiangchun vs. Cshiqishi) chicken populations. Using the BSC for individual and pairwise populations, the Nei's standard genetic distances between the chicken populations were determined and ranged from 0.0043 (Jiangchun vs. Cshiqishi) to 0.1375 (Rugao vs. Cshiqishi). The reconstructed dendrogram linked the Jiangchun and Cshiqishi chickens as closely related populations, followed by Wan-Nan, while the Rugao was the most genetically distant among the populations.

• Parasites, Hosts and Diseases
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• 제35권4호
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• pp.295-298
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• 1997

요코가와흡충과 미야타흡충의 genomic DNA를 RAPD 분석을 이용하여 비교하였다. 상업적으로 구 입한 60-70%의 G+C 성분을 가진 무작위 10-mer oligonucleotide 표지자 (Kit A, Operon Technologies Inc., CalifDmia, USA) 20개 중에서 다음의 8개를 이용하여 두 홉충간에 구별이 가능한 밴드양상을 관찰할 수 있었다: OPA-02,5-TCCCGAGCTG-3; OPA-09,5-GGGTAACGCC-3; OPA-10, 5-GTGATCGCAG-3; OPA-11, 5-CAATCGCCGT-3; OPA-13, 5-CAGCACCCAC-3; OPA-17, 5-GACCGCTGT-3; OPA-19,5-CAAACGTCGG-3; OPA-20, 5-GTrCCGATCC-3. 이 연구의 결과로 미야타흡충은 요코가와흡충과 서로 다른 유전자 염기 서열을 가지고 있음이 암시 되었다.

• Mycobiology
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• 제38권1호
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• pp.17-25
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• 2010

The common split-gilled mushroom, Schizophyllum commune is found throughout the world on woody plants. This study was initiated to evaluate conditions for favorable vegetative growth and to determine molecular phylogenetic relationship in twelve different strains of S. commune. A suitable temperature for mycelial growth was obtained at $30^{\circ}C$. This mushroom grew well in acidic conditions and pH 5 was the most favorable. Hamada, glucose peptone, Hennerberg, potato dextrose agar and yeast malt extract were favorable media for growing mycelia, while Lilly and glucose tryptone were unfavorable. Dextrin was the best and lactose was the less effective carbon source. The most suitable nitrogen sources were calcium nitrate, glycine, and potassium nitrate, whereas ammonium phosphate and histidine were the least effective for the mycelial growth of S. commune. The genetic diversity of each strain was investigated in order to identify them. The internal transcribed spacer (ITS) regions of rDNA were amplified using PCR. The size of the ITS1 and ITS2 regions of rDNA from the different strains varied from 129 to 143 bp and 241 to 243 bp, respectively. The sequence of ITS1 was more variable than that of ITS2, while the 5.8S sequences were identical. A phylogenetic tree of the ITS region sequences indicated that the selected strains were classified into three clusters. The reciprocal homologies of the ITS region sequences ranged from 99 to 100%. The strains were also analyzed by random amplification of polymorphic DNA (RAPD) with 20 arbitrary primers. Twelve primers efficiently amplified the genomic DNA. The number of amplified bands varied depending on the primers used or the strains tested. The average number of polymorphic bands observed per primer was 4.5. The size of polymorphic fragments was obtained in the range of 0.2 to 2.3 kb. These results indicate that the RAPD technique is well suited for detecting the genetic diversity in the S. commune strains tested.

• Asian-Australasian Journal of Animal Sciences
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• 제19권9호
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• pp.1234-1239
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• 2006

Murrah and NiliRavi are the important North Indian buffalo breeds occupying the prominent position of being the highest milk producers. These breeds are more or less similar at morphological as well as physiological levels. The technique of RAPD-PCR was applied in the present study to identify a battery of suitable random primers to detect genetic polymorphism, elucidation of the genetic structure and rapid assessment of the differences in the genetic composition of these two breeds. A total of 50 random primers were screened in 24 animals each of Murrah and NiliRavi buffaloes to generate RAPD patterns. Of these, 26 (52%) primers amplified the buffalo genome generating 263 reproducible bands. The number of polymorphic bands for the 26 chosen RAPD primers varied from 3 (OPG 06 and B4) to 26 (OPJ 04) with an average of 10.1 bands per primer and size range of 0.2 to 3.2 kb. DNA was also pooled and analyzed to search for population specific markers. Two breed specific RAPD alleles were observed in each of Murrah (OPA02 and OPG16) and NiliRavi (OPG09) DNA pools. RAPD profiles revealed that 11 (4.2%) bands were common to all the 48 individuals of Murrah and NiliRavi buffaloes. Pair-wise band sharing calculated among the individual animals indicated considerable homogeneity of individuals within the breeds. Within breed, band sharing values were relatively greater than those of interbreed values. The low genetic distance (Nei's) value (0.109) estimated in this study is in accordance with the origin and geographical distribution of these breeds. The RAPD analysis indicated high level of genetic similarity between these two important North Indian buffalo breeds.

• 생명과학회지
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• 제20권10호
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• pp.1538-1543
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• 2010

본 연구는 우리나라 겨자 17집단에 대한 유전적 다양도와 집단구조를 조사하였다. 60개의 다형성 좌위와 18개 단형성 좌위가 발견되었다. 다형성 밴드의 비율은 전남 진도 집단이 가장 높았으며 재배종이 가장 낮았다. 대립유전자좌위의 수는 1.221이였으며 유효한 대립유전자좌위의 수는 1.167이였다. 이 종의 전형적인 집단은 작고 격리되어 낮은 유전적 다양도를 가지고 있었다. 전체 다양도는 0.347이였으며 집단 내 다양도는 0.141이였다. 집단간분화를 나타내는 척도는 0.589였다. 아는 58.9%의 다양도가 집단간에 있음을 시사한다. 세대 간 이주하는 개체수는 0.617로 낮았다. RAPD는 겨자 집단을 구분하는데 유익하였다.

• International Journal of Industrial Entomology and Biomaterials
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• 제29권2호
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• pp.169-178
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• 2014

In this study, we investigated genetic diversity of 22 microsporidian isolates infecting tropical tasar silkworm, Antheraea mylitta collected from various geographical forest locations in the state of Jharkhand, India, using polymerase chain reaction (PCR)-based marker assay: random amplified polymorphic DNA (RAPD). A type species, NIK-1s_mys was used as control for comparison. The shape of mature microsporidians was found to be oval to elongate, measuring 3.80 to $5.10{\mu}m$ in length and 2.56 to $3.30{\mu}m$ in width. Of the 20 RAPD primers screened, 16 primers generated reproducible profiles with 298 polymorphic fragments displaying high degree of polymorphism (97%). A total of 14 RAPD primers produced 45 unique putative genetic markers, which were used to differentiate the microsporidians. Calculation of genetic distance coefficients based on dice coefficient method and clustering with un-weighted pair group method using arithmetic average (UPGMA) analysis was conducted to unravel the genetic diversity of microsporidians infecting tasar silkworm. The similarity coefficients varied from 0.059 to 0.980. UPGMA analysis generated a dendrogram with four microsporidian groups, which appear to be different from each other as well as from NIK-1s_mys. Two-dimensional distribution based on Euclidean distance matrix also revealed considerable variability among different microsporidians identified from the tasar silkworms. Clustering of few microsporidian isolates was in accordance with the geographic origin. The results indicate that the RAPD profiles and specific/unique genetic markers can be used for differentiating as well as to identify different microsporidians with considerable accuracy.

• 한국양식학회지
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• 제17권4호
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• pp.251-257
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• 2004

상업적 가치가 높고 양식 가능성이 있는 백합과 5종을 대상으로 RAPD방법을 이용한 개체간의 유전적 유연관계를 조사하였다. DNA 추출은 protein K-phenol방법을 사용하여 각 종의 후폐각근에서 추출하였다. RAPD-PCR결과 15개의 primer들이 증폭되었고, 그들로부터 각각 1-3개의 band를 볼 수 있었다. 백합과 5종간 유전적 유사도는 살조개와 바지락간에는 0.84, 개조개와 백합간에는 0.87로 각각의 두 종간에는 놓은 유사도를 보였고, 가무락조개는 살조개와 바지락간에 0.78의 유사도를 보였으며, 개조개와 백합, 살조개, 바지락, 가무락조개간에는 0.46의 비교적 낮은 유전적 유사도를 보였다. RAPD 방법으로 종내개체 유전변이를 파악하기는 힘들어도, 양식이나 자원증식을 위한 패류의 우수형질 선택에 있어 기본 정보를 제공 할 수는 있을 것이다. 또한, 살조개 대량종묘생산의 방법은 유전적 유사도가 가장 가까운 바지락을 중심으로 이루어져야 할 것으로 사료된다.

• Parasites, Hosts and Diseases
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• 제39권1호
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• pp.43-48
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• 2001

We have studied the genetic differences among four isolates of Trichinella including a new strain of Trichinella spiralis (ISS 623) recently found from a human case who took a badger in Korea. Because they have a different host origin and came from geographically separated regions, we supposed the genetic pattern of the isolates might be different as had been previously reported. It was analysed by PCR-RFLP analysis of the rDNA repeat that can readily distinguish a species or strain from others. Isolated genomic DNA of each isolate of Trichinella larvae was amplified with ITSl specific primers and digested with restriction endonucleases. The PCR product of ITSl was confirmed using Southern blot analysis to be a 910 Up fragment. The restriction fragments of each isolate had variable patterns when it was digested with Rsa I only. According to the RFLP patterns, the estimated genetic divergence between each isolate was different. In conclusion, four isolates of Thichinella including a new strain of T. spiralis obtained from a Korean patient may have genetic differences in the ITSl region and the Shanghai isolate was genetically more similar to the Japanese unknown isolate than others in the ITSl region.

• The Plant Pathology Journal
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• 제29권4호
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• pp.357-363
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• 2013

The fungus Venturia nashicola is the causal agent of scab on Asian pears. For the rapid and reliable identification as well as sensitive detection of V. nashicola, a PCR-based technique was developed. DNA fingerprints of three closely related species, V. nashicola, V. pirina, and V. inaequalis, were obtained by random amplified polymorphic DNA (RAPD) analysis. Two RAPD markers specific to V. nashicola were identified by PCR, after which two pairs of sequence characterized amplified region (SCAR) primers were designed from the nucleotide sequences of the markers. The SCAR primer pairs, designated as D12F/D12R and E11F/E11R, amplified 535-bp and 525-bp DNA fragments, respectively, only from genomic DNA of V. nashicola. The specificity of the primer sets was tested on strains representing three species of Venturia and 20 fungal plant pathogens. The nested PCR primer pair specific to V. nashicola was developed based on the sequence of the species-specific 525-bp DNA fragment amplified by primer set E11F/E11R. The internal primer pair Na11F/Na11R amplified a 235-bp fragment from V. nashicola, but not from any other fungal species tested. The nested PCR assay was sensitive enough to detect the specific fragment in 50 fg of V. nashicola DNA.

• 생명과학회지
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• 제26권4호
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• pp.406-413
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• 2016

전복은 전복속(Haliotis)에 속하며 전 세계적으로 식품산업에서 중요한 복족류 연체동물이다. 우리나라에는 6개종; 북방전복(Haliotis discus hannai), 둥근전복(Haliotis discus discus), 왕전복(Haliotis madaka), 말전복(Haliotis gigantea), 오분자기(Haliotis diversicolor diversicolor), 마대오분자기(Haliotis diversicolor supertexta)가 보고되어 있다. 이 연구에서는 우리나라 해역에 서식하는 중ㆍ대형 전복과 4종인 북방전복, 둥근전복, 왕전복, 말전복의 유전학적 유연관계를 분석하기 위하여 미토콘드리아의 cytochrome c oxidase subunit I (COI) 유전자와 Random Amplified Polymorphic DNA (RAPD) 분석법을 실시 하였다. 본 연구의 결과 COI 유전자 분석과 RAPD 분석을 활용하면 4종의 전복 중 북방전복, 둥근전복, 왕전복을 한 그룹으로 나머지 한 그룹을 말전복으로 구분하는 종 분류는 명확히 구분할 수 있었다. 이러한 결과는 전복 교잡육종을 이용한 수출용 전복 신종자 개발에 있어 주요 대상종인 전복과 4종에 대한 유전적 근연 관계를 규정함으로써 향후 교잡육종 연구의 기초 자료를 제공할 수 있을 것으로 사료된다.