• Parasites, Hosts and Diseases
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• 제47권4호
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• pp.369-375
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• 2009

Diphyllobothrium nihonkaiense was first described by Yamane in 1986 but the taxonomical features have been obscure due to lack of critical morphologic criteria in its larval and adult stages. In Korea, this tapeworm had long been known as Diphyllobothrium latum. In this study, we observed 62 specimens collected from Korean residents and analyzed them by morphological features and nucleotide sequences of mitochondrial cox1 gene as well as the ITS1 region. Adult tapeworms were examined after carmine or trichrome stain. Longitudinal sections of the gravid proglottids showed an obtuse angle of about 150 degree between the cirrus sac and seminal vesicle. This angle is known as a major differential point compared with that of D. latum. Nucleotide sequence differences between D. latum and the specimens from Koreans represented 17.3% in mitochondrial DNA cox1 gene. Sequence divergence of ITS1 among 4 Korean isolates was 0.3% and similarity was 99.7% with D. nihonkaiense and D. klebanovskii. All of the Korean specimens analyzed in this study were identified as being D. nihonkaiense (n = 62). We propose its Korean name as "Dong-hae-gin-chon-chung" which means 'long tapeworm of the East Sea' for this newly analyzed diphyllobothriid tapeworm in Korea.

• Animal Systematics, Evolution and Diversity
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• 제9권2호
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• pp.171-179
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• 1993

한반도의 8개 지역에서 채집한 39마리의 등줄쥐(Apodemus agrarius coreae Thomas)의 미토콘드리아 DNA(mtDNA)를 8가지의 제한효소로 digestion시켰으며, 이들 mtDNA 절단단편들을 분석하였다. 전부 31개의 절단단편들이 나타났고, 7개의 mtDNA clone이 밝혀졌으며, 전체 8개 지역의 39마리 중에서 8개 지역의 32마리(속초, 1중에 1; 치악산, 5중에 4; 월악산, 3중에 3; 속리산, 2중에 2; 덕유산, 2중에 2; 지리산, 4중에 3; 해남, 4중에 2; 청주, 18중에 15)가 동일한 한 clone에 속했다. 또한 이들 7개 mtDNA clone들 간의 nucleotide-sequence divergence (p)의 범위는 0.2%-2.3%였으며, 이들 clone들은 뚜렷한 차이를 보이는 subgroup으로 구분되지 않았다. 한반도산 등줄쥐는, mtDNA genotype에 있어서도 뚜렷한 차이를 보이는 집단으로 나뉘어지지 않았으므로, 단일 아종인 Apodemus agrarius coreae임을 확인하게되었다.

• 대한본초학회지
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• 제28권5호
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• pp.1-6
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• 2013

Objectives : This study was conducted to identify the original Sinomini Caulis et Rhizoma plant among Stephania tetrandra, Cocculus trilobus, and Aristolochiae fangchi to develop the genetic marker for Sinomini Caulis et Rhizoma. Methods : Sinomenium acutum was identified by the classification and identification committee of the National Center for Standardization of Herbal Medicines. The chloroplast ndhF gene was amplified. We performed sequences alignment analysis of Sinomenium acutum, Stephania tetrandra, C. trilobus, and A. fangchi using BioEdit program. The SFR markers designed were consisted of SF01, SR04, and SR05 primers. Results : Many variations of Sinomeni Caulis et Rhizoma are currently commercialized as herbal medicine. We compared the base sequences of the ndhF intergenic space of chloroplast DNA with Sinomenium acutum, Stephania tetrandra, C. trilobus, and A. fangchi. According to the results, it showed that the nucleotide variations were seen in 30 genes of four species. Phylogenetic analysis revealed that 4 species were classified into five groups based on an inter-group divergence in nucleotide sequence of 9%. We developed SFR marker nucleotides enough to authenticate respective species and confirmed its application on the band size at 419 base pair. These sequence differences at corresponding positions were available genetic markers to identity the Sinomeni Caulis et Rhizoma. Conclusions : Base on these results, the ndhF region was effective in distinguishing Sinomini Caulis et Rhizoma The SFR genetic marker was useful for identifying Sinomini Caulis et Rhizoma with other species.

• 식물병연구
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• 제28권2호
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• pp.98-107
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• 2022

The incidence of Tomato brown rugose fruit virus (ToBRFV) and biological and molecular characterization of the Palestinian isolates of ToBRFV are described in this study. Symptomatic leaf samples obtained from Solanum lycopersicum L. (tomatoes) and Nicotiana tabacum L. (cultivated tobacco) plants were tested for tobamoviruses infection by reverse transcription polymerase chain reaction. Tomato leaf samples collected from Tulkarm and Qalqilia are infected with ToBRFV-PAL with an infection rate of 76% and 72.5%, respectively. Leaf samples collected from Jenin and Nablus were found to be mixed infected with ToBRFV-PAL and Tobacco mosaic virus (TMV) (100%). Sequence analysis of the ToBRFV-PAL genome showed that the net average nucleotide divergence between ToBRFV/F48-PAL strain and the Israeli and Turkish strains was 0.0026398±0.0006638 (±standard error of mean), while it was 0.0033066±0.0007433 between ToBRFV/F42-PAL and these two isolates. In the phylogenetic tree constructed with the complete genomic sequence, all the ToBRFV isolates were clustered together and formed a sister branch with the TMV. The sequenced Palestinian isolates of ToBRFV-PAL shared the highest nucleotide identity with the Israeli ToBRFV isolate suggesting that the virus was introduced to Palestine from Israel. The findings of this study enhance our understanding of the biological and molecular characteristics of ToBRFV which would help in the management of the disease.

• 미생물학회지
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• 제44권3호
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• pp.212-220
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• 2008

본 연구는 Prevotella nigrescens ATCC $33563^T$에 대한 균주 특이 DNA 프로브라고 보고된 Pn10 프로브의 균주 특이성을 한국인에서 분리된 P. nigrescens의 임상분리 균주를 이용하여 검증하고, P. nigrescens ATCC $33563^T$ 균주 특이 PCR 프라이머를 개발하고자 시행되었다. P. nigrescens와 유전학적으로 가장 가까운 Prevotella intermedia를 포함한 구강 내 치주질환 원인균종인 5균종의 표준균주 및 참고균주, 그리고 P. nigrescens와 P. intermedia의 임상분리 균주를 이용하여 Southern blot 분석법을 시행하였다. Southern blot 분석 결과 Pn10 DNA 프로브에 P. nigrescens ATCC $33563^T$ 및 ChDC KB6 두 균주 지놈 DNA가 검출되었다. P. nigrescens KB6 균주에서 Pn10 DNA 프로브와 상동성이 있는 부위를 PCR법으로 증폭(KB6-Pn10)하여 클로닝한 다음 Pn10 DNA프로브와 같이 핵산 염기서열을 결정하여 상동성을 비교하였다. 그 결과 Pn10과 KB6-Pn10의 핵산염기서열간의 Percent identity는 98.8%였으며, divergence는 0.6%였다. Pn10 DNA 프로브의 핵산염기서열을 바탕으로 두 중류 프라이머 쌍(Pn10-F-AC/Pn10-R-AC 및 Pn10-F-A/Pn10-R-A)을 설계 및 제작하여 P. nigrescens ATCC $33563^T$에 대한 균주 특이성을 PCR법으로 검증하였다. 이들 프라이머 쌍들의 민감도(sensitivity) 조사 결과, 이들은 P. nigrescens ATCC $33563^T$ 지놈 DNA 4 pg까지 검출할 수 있음을 알았다. 이상의 연구 결과를 종합하면, Pn10 DNA 핵산염기서열을 바탕으로 설계된 Pn10-F-AC/Pn10-R-AC 및 Pn10-F-A/Pn10-R-A 프라이머 쌍들은 P. nigrescens ATCC $33563^T$를 신속 정확하게 검출하는 수 있어, 균주의 보존적 측면에서 유용하게 이용될 수 있을 것으로 생각된다.

• 한국응용곤충학회지
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• 제56권4호
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• pp.377-385
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• 2017

멸구과(Delphacidae) 8종의 internal transcribed spacer 2 (ITS2) DNA 염기서열로 종간 차이 추정값을 비교하고 분자계통수를 추론하였다. ITS2 DNA 염기서열 길이는 종(species)마다 550 bp (흰등멸구)에서 699 bp (겨풀멸구)까지 차이를 보였다. 같은 Nilaparvata 속의 겨풀멸구와 벼멸구붙이 사이의 염기서열 차이 추정값($d{\pm}S.E.$)은 $0.001{\pm}0.001$로 가장 낮았으며, 사슴멸구와 일본멸구 사이는 $0.579{\pm}0.021$로 가장 높았다. 벼멸구와 다른 멸구류들과의 종간 염기서열 차이 추정값은 $0.056{\pm}0.008$ (겨풀멸구)에서부터 $0.548{\pm}0.021$ (일본멸구)로 구분되었다. 반면, Neighbor-joining 방법으로 추론된 분자계통수에서는 겨풀멸구와 벼멸구붙이를 제외하고 나머지 멸구류들은 독립된 다른 그룹으로 분지되었다. 벼멸구의 ITS2 염기서열을 참고하여 벼멸구 특이 고리매개등온증폭(loop-mediated isothermal amplification, LAMP) 프라이머 4 세트(BPH-38, BPH-38-1, BPH-207 및 BPH-92)를 제작하였다. 이들 각각을 벼멸구, 흰등멸구 및 애멸구의 게놈 DNA와 $65^{\circ}C$에서 60분간 반응시켰을 때, 벼멸구 시료에서만 증폭 산물들이 관찰되었다. BPH-92 LAMP 프라이머 세트로 $65^{\circ}C$에서 벼멸구 DNA의 양(0.1 ng, 1 ng, 10 ng, 100 ng)과 반응시간(20분, 30분, 40분, 60분)을 달리하여 형광반응을 관찰하였을 때, 20분과 30분 반응에서는 100 ng 까지에서도 발광여부 구별이 어려웠다. 그러나 40분 반응에서는 10 ng 이상에서, 60분 반응에서는 0.1 ng 이상에서 발광여부가 명확히 구별되었다.

• 생명과학회지
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• 제12권6호
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• pp.821-834
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• 2002

알렉산드륨 적조생물의 리보소옴 알엔에이 유전자의 ITS1, 2 및 5.8S부위를 대상으로 종간 혹은 종내의 유전적 다양도를 조사하기 위하여 지리적으로 격리된 33 스트레인 유전자의 염기서열를 비교했다. 진해만에서 분리된 AT-2, AT-6, AT-10, AT-A, AT-B 5클론은 일본종 OFX151-A과 동일한 유전자임을 발견했다. ITS부위에서 가장 짧은 종은 A. margalefi로 481 bp이며 가장 긴 종은 A. affine으로 528 bp로 나타났다. ITS1과 ITS2 염기서열에 대한 상호관계는 역으로 나타낸 반면에, G＋C 함량에 대한 상호관계는 플러스로 나타났다. 유전적 변이율은 0.3% (1 bp)에서 53% (305 bp)였다. A. tamarense과 가장 적게 유전적 변이율을 보인 종은 A. fundyense(1.2-2.3% ＝ 6-12 bp)인 반면에, A. catenella와는 큰 변이율 (19.8% ＝ 102 bp)을 보였고, A. catenella와 A. fundyense은 19.7% 상이하였다. 알렉산드륨 적조생물의 bootstrap은 약하게 지지되는 데도 불구하고, A. catenella 분리종은 독립적인 그룹으로 형성하여 상호간에는 강력한 bootstrap 값은 PAUP과 NJ 분석에서 보였다. A. cohorticula와 A. frateculus 적조생물은 항상 sub-group 내에서 높은 bootstrap을 가졌다. 결론적으로 ITS부위의 염기서열 분석은 알렉산드륨 적조생물의 집단내 혹은 집단간의 계통분류을 밝히는데 유용한 것으로 보였다.

• Parasites, Hosts and Diseases
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• 제42권4호
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• pp.159-167
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• 2004

Sequences of the complete protein-coding portions of the mitochondrial (mt) genome were analysed for 6 species of cestodes (including hydatid tapeworms and the pork tapeworm) and 5 species of trematodes (blood flukes and liver- and lung-flukes). A near-complete sequence was also available for an additional trematode (the blood fluke Schistosoma malayensis). All of these parasites belong to a large flatworm taxon named the Neodermata. Considerable variation was found in the base composition of the protein-coding genes among these neodermatans. This variation was reflected in statistically-significant differences in numbers of each inferred amino acid between many pairs of species. Both convergence and divergence in nucleotide, and hence amino acid, composition was noted among groups within the Neodermata. Considerable variation in skew (unequal representation of complementary bases on the same strand) was found among the species studied. A pattern is thus emerging of diversity in the mt genome in neodermatans that may cast light on evolution of mt genomes generally.

• 대한미생물학회지
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• 제34권6호
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• pp.561-571
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• 1999

Diagnosis of mycobacterial infection is dependent upon the isolation and identification of causative agents. The procedures involved are time consuming and technically demanding. To improve the laborious identification process mycobacterial systematics supported by gene analysis is feasible, being particularly useful for slowly growing or uncultivable mycobacteria. To complement genetic analysis for the differentiation and identification of mycobacterial species, an alternative marker gene, rpoB encoding the ${\beta}$ subunit of RNA polymerase, was investigated. rpoB DNAs (342 bp) were amplified from 52 reference strains of mycobacteria including Mycobacterium tuberculosis H37Rv (ATCC 27294) and clinical isolates by the PCR. The nucleotide sequences were directly determined (306 bp) and aligned using the multiple alignment algorithm in the MegAlign package (DNASTAR) and MEGA program. A phylogenetic tree was constructed with a neighborhood joining method. Comparative sequence analysis of rpoB DNA provided the basis for species differentiation. By being grouped into species-specific clusters with low sequence divergence among strains belonging to same species, all the clinical isolates could be easily identified. Furthermore RFLP analysis enabled rapid identification of clinical isolates.

• Parasites, Hosts and Diseases
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• 제54권3호
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• pp.369-373
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• 2016

The 2 species of the genus Anoplocephala (Anoplocephalidae), A. perfoliata and A. magna, are among the most important equine cestode parasites. However, there is little information about their differences at the molecular level. The present study revealed that the mitochondrial (mt) genome of A. magna was 13,759 bp in size and 700 bp shorter than that of A. perfoliata. The 2 species includes 2 rRNA, 22 tRNA, and 12 protein-coding genes each. The size of each of the 36 genes was the same as that of A. perfoliata, except for cox1, rrnL, trnC, trnS2(UCN), trnG, trnH, trnQ, and trnP. In the full mitochondrial genome, the sequence similarity was 87.1%. The divergence in the nucleotide and amino acid sequences of individual protein-coding genes ranged from 11.1% to 16% and 6.8% to 16.4%, respectively. The 2 non-coding regions of the mt genome of A. magna were 199 bp and 271 bp in length, while the equivalent regions in A. perfoliata were 875 bp and 276 bp, respectively. The results of this study support the proposal that A. magna and A. perfoliata are separate species, consistent with previous morphological analyses.