• 한국양식학회:학술대회논문집
• /
• 한국양식학회 2003년도 추계학술발표대회 논문요약집
• /
• pp.134-135
• /
• 2003

The geographic expansion of the toxic dinoflagellates genus Alexandrium has been shown to be world wide ranging. The members of the genus Alexandrium ocnstituted of 20-30 species did not show substantial differences in their morphology, which is mostly referred in the 'tamarensis species complex', except some species. Though rDNA sequences variations are very few and pseudogene types are so diverse that it is difficult to use them as the specific markers. In this study, we outlined Korean and Japanese A, tamarense and A. catenella regional isolates by phylogenetic analysis inferred from no cutting alignments of LSU rDNA D1-D2 and SSU rDNA sequences to group these regional isolates. The results were compared to RFLP patterns of PCR products targeted chloroplast DNA. Lastly screening of highly repeated microsatellite DNA which is frequently used for population analysis in eukaryotes was conducted. A. catenella regional strains identified by the sequencing of rDNA D1-D2 domain were divided into at least 3 groups of type E, CMC and Chinese type, divergence root may not be deep comparing with that of A. tamarense whose pseudogenes are very variable. Results of RFLP pattern and the phylogeny of the unknown gene targeting chloroplast showed that Korean and Japanese A. catenella regional isolates were divided into 3 types: Korean, Japanese and the third CMC types. Population-specific PCR amplification with Japanese A. catenella type-specific PCR primers was useful method for population analysis of A. catenella. Various types of satellite sequences such as 5 nucleotides repeats were obtained from A. tamarense and A. catenella. The 5 nucleotides repeats were primed at the both 3'and 5' ends, and these repeats were prominent as longer repeated motifs. This repeated DNA was intercalated as internal sequences containing various types subrepeats. It is expected that these satellite DNA would be a useful molecular population marker through detail comparison among Alexandrium regional isolates to trace their transferring pathway and to prevent their human-associated their regional extents.

• Asian-Australasian Journal of Animal Sciences
• /
• 제18권12호
• /
• pp.1691-1695
• /
• 2005

The bovine lymphocyte antigen (BoLA)-DRB3 gene encodes cell surface glycoproteins that initiate immune responses by presenting processed antigenic peptides to CD4 T helper cells. DRB3 is the most polymorphic bovine MHC class II gene which encodes the peptide-binding groove. Since different alleles favour the binding of different peptides, DRB3 has been extensively evaluated as a candidate marker for associations with various bovine diseases and immunological traits. For that reason, the genetic diversity of the bovine class II DRB3 locus was investigated by polymerase chain reaction-restriction fragment length polymorphism method (PCR-RFLP). This study describes genetic variability in the BoLA-DRB3 in Iranian Golpayegani Cattle. Iranian Golpayegani Cows (n = 50) were genotyped for bovine lymphocyte antigen (BoLA)-DRB3.2 allele by polymerase chain reaction and restriction fragment length polymorphism method. Bovine DNA was isolated from aliquots of whole blood. A two-step polymerase chain reaction followed by digestion with restriction endonucleases RsaI, HaeIII and BstYI was conducted on the DNA from Iranian Golpayegani Cattle. In the Iranian Golpayegani herd studied, we identified 19 alleles.DRB3.2${\times}$16 had the highest allelic frequency (14%), followed by DRB3.2${\times}$7 (11%). Six alleles (DRB3.2${\times}$25, ${\times}$24, ${\times}$22, ${\times}$20, ${\times}$15, ${\times}$3) had frequencies = 2%. Although additional studies are required to confirm the present findings, our results indicate that exon 2 of the BoLA-DRB3 gene is highly polymorphic in Iranian Golpayegani Cattle.

• 한국해양학회지:바다
• /
• 제15권1호
• /
• pp.25-35
• /
• 2010

플랑크톤의 종다양성은 특정 지역의 수계환경 변화 모니터링에 있어 중요 생태지표로써, 환경 평가에 유용한 정보로 사용되고 있다. 기존의 종다양성 평가는 주로 형태학적 형질에 근거한 종동정을 통해 이루어졌으나, 많은 시간과 전문성을 필요로 하고 연구자의 주관적 판단에 의존하는 단점이 있다. 따라서, 본 연구에서는 채수된 환경시료에 대해 보다 빠르고 정확한 플랑크톤 종다양성을 파악하기 위하여 분자마커를 활용한 분자모니터링 기법을 도입하였다. 서낙동강(김해교)과 남해 연얀(남해도) 정점에서 각각 채수된 환경시료에서 DNA를 추출한 후 18S nuclear ribosomal RNA 유전자를 대상으로 중합효소연쇄반응을 수행하였다. 클로닝 과정을 통해 만들어진 각각의 클론 라이브러리에서 클론을 무작위로 선택하여 제한효소절편다형성 패턴분석을 한 후 특이성을 가지는 클론을 선별하였다. 김해교에서는 60개 블론을 대상으로 44개의 특이적 클론을 선별하였고 남해에서는 150개 클론을 대상으토 27개의 클론을 선별하였다. 이틀 클론틀에 대한 염기서열 분석결과 다양한 계통분류군에 속승하는 플랑크톤의 종조성 결과를 보여주었다(김해교: Heterokontophyta(7), Ciliophora(23), Dinophyta(l), Chytridiomycota(l), Rotifera(I), Arthropoda (11), 남해: Ciliophora( 4), Dinophyta(3), Crγptophyta(l)，Arthropoda(19)). 본 연구를 통하여 분자마커를 활용한 분자모니터링 기법이 기존 형태학적 형질에 근거한 분석이 가지는 한계를 보완하여 채수된 환경시료의 종조성 분석에 효율적으로 사용될 수 있다고 판단된다.

• 한국응용곤충학회지
• /
• 제46권2호
• /
• pp.175-182
• /
• 2007

국내 서식하는 복숭아순나방 (Grapholita molesta (Busck))과 복숭아심식나방 (Carposina sasakii (Matsumura))의 유충은 사과 과실내부를 섭식하여 피해를 주는 해충이다. 사과를 수출할 때 복숭아심식나방은 수출대상국들로부터 검역 대상해충이다. 반면에 복숭아순나방은 광범위한 분포로 비교적 수입국으로부터 검역 대상 해충은 아니지만, 사과 과실 내부에서 발견되는 경우 복숭아심식나방으로 오인될 수 있다. 이는 발견되는 유충을 가지고 형태적으로 두 종을 구분하기 어렵기 때문이다. 특별히 수입국 검역단계에서 이러한 불완전한 동정 실태는 수출 사과의 폐기 또는 반송과 수출중단 등과 같은 막대한 경제적 손실을 초래하게 된다. 이에 이들을 구분할 수 있는 분자지표 개발이 요구되었다. 두 종의 미토콘드리아 DNA를 대상으로 다형을 보이는 여러 영역의 염기서열을 분석하였다. 이 서열을 바탕으로 진단용 제한위치가 결정되고 종 특이적 프라이머가 제작되었다. 본 연구는 세 부위의 종 특이적 제한효소 위치에 따라 PCR-RFLP 기술과 종 특이적 프라이머를 이용하여 진단용 PCR 기술을 개발하였다.

• Molecules and Cells
• /
• 제26권3호
• /
• pp.314-318
• /
• 2008

Korean long-tailed goral (Nemorhaedus caudatus) is one of the most endangered species in South Korea. However, detailed species distribution and sex ratio data on the elusive goral are still lacking due to difficulty of identification of the species and sex in the field. The primary aim of this study was to develop an economical PCR-RFLP method to identify species using invasive or non-invasive samples from five Korean ungulates: goral (N. caudatus), roe deer (Capreolus pygargus), feral goat (Capra hircus), water deer (Hydropotes inermis) and musk deer (Moschus moschiferus). The secondary aim was to find more efficient molecular sexing techniques that may be applied to invasive or non-invasive samples of ungulate species. We successfully utilized PCR-RFLP of partial mitochondrial cytochrome b gene (376 bp) for species identification, and sex-specific amplification of ZFX/Y and AMELX/Y genes for sexing. Three species (goral, goat and water deer) showed distinctive band patterns by using three restriction enzymes (Xbal, Stul or Sspl). Three different sexing primer sets (LGL331/335 for ZFX/Y gene; SE47/48 or SE47/53 for AMELX/Y gene) produced sex-specific band patterns in goral, goat and roe deer. Our results suggest that the molecular analyses of non-invasive samples might provide us with potential tools for the further genetic and ecological study of Korean goral and related species.

• 대한한의학회지
• /
• 제20권4호
• /
• pp.11-15
• /
• 2000

Radix Angelicae Gigantis is sweet and pungent in flavor, warm in property. Its effects are tonifying the blood, promoting blood circulation, relieving pain and moistening the bowels. Its indications are blood deficiency syndrome characterized by sallow complexion, dizziness, irregular menstruation, amenorrhea, pains due to blood stasis, and rheumatic arthralgia. Using genes of A. gigas, A. acutiloba, and A. sinensis, the origin of which is identified, as criteria, we analysed many kinds of Angelica with RAPD and RFLP on ITS region, in order to compare and discriminate genes extracted from crude drugs ‘Dang-gui’, that are produced in Korea on the one hand and imported on the other hand. We reached the following conclusion. 1. We could extract DNA from both original plant and dried plant. 2. Especially Uniprimer #1, Uniprimer #2, Uniprimer #4 and Uniprimer ＃9 were useful. 3. Among the restriction enzymes Sma I, Msp I, Hae III, and Hinf I, used in this experiment, four restriction enzymes except Hinf I could be used properly in discriminating all samples used as A. gigas. We think that this result can be used as a method of discriminating crude drug of Angelica L. related drugs, and used in controlling quality and circulation.

• 한국해양학회지:바다
• /
• 제17권3호
• /
• pp.160-180
• /
• 2012

본 연구는 메타게놈 분석법을 기초로 낙동강 하류 담수 환경의 물금과 기수 환경의 을숙도대교 정점에서 채수된 환경 시료내의 진핵 플랑크톤 종다양성 및 군집 구조를 비교 분석하고자 하였다. 수환경 시료에서 추출된 DNA에 대한 environmental Polymerase Chain Reaction(PCR)을 수행하여 18S rDNA 클론라이브러리를 구축하였고, colony PCR, PCR-Restriction Fragment Length Polymorphism(RFLP), 염기서열 결정 및 유사도 분석을 통하여 종다양성을 분석하였다. 물금 및 을숙도대교 정점에서 338개의 클론들을 분석하였고(170 clones, 물금; 168 clones, 을숙도대교), 그 결과 총 74개의 phylotype을 발굴하였다(49개, 물금; 25개, 을숙도대교). 발굴된 phylotype에 대한 계통 분석 결과, Stramenopiles, Cryptophyta, Viridiplantae, Alveolata, Rhizaria, Metazoa 및 Fungi 등의 분류군에 속하는 다양한 생물종이 발굴되었으며, 국내 미기록종 및 신종 후보 가능 생물종과 속(genus)이상의 새로운 분류학적 처리가 필요한 생물종의 존재를 확인하였다. 특히 Stramenopiles의 Pirsonia 및 Alveolata의 Perkinsea에 속하는 phylotypes 등 국내 미기록 생물종을 포함한 숨은 종다양성(cryptic species diversity)의 발굴은 분자모니터링 기법이 낙동강 하구역 수생태계 변화 모니터링을 위한 새로운 유용한 생물학적 정보를 제공할 수 있음을 제시하고 있다.

• Asian-Australasian Journal of Animal Sciences
• /
• 제21권4호
• /
• pp.465-470
• /
• 2008

This study describes genetic variability in the BoLA-DRB3 gene in Iranian native cattle (Bos Indicus and Taurus) and relationships between these breeds. This is the first study of genetic polymorphism of the BoLA-DRB3 gene in Iranian native cattle. We examined exon 2 of the major histocompatibility complex (MHC) class II DRB3 gene from 203 individuals in four populations of Iranian native cattle (52 Sarabi, 52 Najdi, 49 Sistani, 50 Golpayegani cattle) using the hemi-nested PCR-RFLP method. We identified the 36 previously reported alleles and one novel pattern (*eac). Analysis of the frequencies of the various BoLA-DRB3.2 alleles in each breed indicated that DRB3.2*52 in Sarabi cattle (23%), DRB3.2 *14 and *24 alleles in Najdi cattle (13%), DRB3.2 *8 allele in Sistani cattle (22%) and DRB3.2*16 allele in Golpayegani cattle (14%), were the most frequent alleles. Allelic frequencies ranged from 1 to 23% among the 36 alleles and there were some alleles that were found only in Iranian cattle. Effective number of alleles in the four breeds was estimated to be 7.86, 11.68, 7.08 and 3.37 in Sarabi, Najdi, Sistani and Golpayegani, respectively. Observed heterozygosities were the highest in Sarabi (94%) and Najdi (94%). A population tree based on the frequency of BoLA-DRB3.2 alleles in each breed suggested that Najdi, Sarabi and Golpayegani cattle clustered together and Najdi and Sarabi were the closest breeds. Sistani cattle differed more from these three breeds. These new data suggest that allele frequencies differ between Iranian cattle breeds.

• 한국가금학회지
• /
• 제35권4호
• /
• pp.335-339
• /
• 2009

초저밀도 아포지단백(Very Low Density Apolipoprotein-II)은 조직의 지방단백질 분비 조성과 밀접한 관련이 있다고 알려진 유전자로서 최근 닭에서 성장 및 체구성과 매우 높은 연관이 있다고 알려져 있다. 본 연구는 닭의 3품종에서 PCR-RFLP 방법을 통해 apoVLDL-II 유전자의 유전자형을 분석하고 Broiler 형(B)과 Fayoumi 형(F)을 결정하였다. 각 품종간 유전자형 빈도는 한국 재래계에서 BB가 0.37, BF가 0.43, FF가 0.2로 검출되었고, 오계에서 BB가 0.2, BF가 0.6, FF가 0.2로 검출되었으며, 백색레그혼에서 BB가 0.16, BF가 0.84로 검출되었다. Broiler 형은 성장이나 체구성을 높이는 방향으로 작용한다는 기존의 보고를 바탕으로 본 연구에서 나타난 집단내 다양한 유전자형 분포는 재검증 작업을 거쳐 육종에 의해 이 형질들을 개량할 수 있음을 의미한다.

• Journal of Microbiology
• /
• 제44권1호
• /
• pp.29-34
• /
• 2006

The purpose of this study was to develop molecular identification method for medical mushrooms and their preparations based on the nucleotide sequences of nuclear large subunit (LSD) rDNA. Four specimens were collected of each of the three representative medicinal mushrooms used in Korea: Ganoderma Incidum, Coriolus versicolor, and Fomes fomentarius. Fungal material used in these experiments included two different mycelial cultures and two different fruiting bodies from wild or cultivated mushrooms. The genomic DNA of mushrooms were extracted and 3 nuclear LSU rDNA fragments were amplified: set 1 for the 1.1-kb DNA fragment in the upstream region, set 2 for the 1.2-kb fragment in the middle, and set 3 for the 1.3-kb fragment downstream. The amplified gene products of nuclear large subunit rDNA from 3 different mushrooms were cloned into E. coli vector and subjected to nucleotide sequence determination. The sequence thus determined revealed that the gene sequences of the same medicinal mushroom species were more than $99.48\%$ homologous, and the consensus sequences of 3 different medicinal mushrooms were more than $97.80\%$ homologous. Restriction analysis revealed no useful restriction sites for 6-bp recognition enzymes for distinguishing the 3 sequences from one another, but some distinctive restriction patterns were recognized by the 4-bp recognition enzymes AccII and HhaI. This analysis was also confirmed by PCR-RFLP experiments on medicinal mushrooms.