• Journal of Ginseng Research
• /
• 제33권3호
• /
• pp.199-205
• /
• 2009

인삼에 대한 microsatellite 개발은 다른 분자적 마커들에 비해 늦게 이루어져, 최근에 와서야 인삼의 microsatellite 들이 보고되고 있는 실정이다. 본 연구에서는, 분리된 microsatellite들 중에서 5 개의 다형성 마커를 선별하여 국내 경작지나 시장에서 유통되는 인삼을 대상으로 유전적 다형성을 조사하고, 각 마커의 특성을 규명하였다. 유전자형 분석은 변성 PAGE와 silver staining법으로 하거나 형광표지 primer로 표지한 PCR 산물을 자동 염기서열 분석기로 분석하였다. 본 연구에서 개발한 5개의 microsatellite 마커들의 평균 대립유전자 수는 3.2 개였으며, 평균 GD는 0.367 였다. 전체적으로 볼 때, PG1419가 가장 높은 다형성을 보였으며 (PIC: 0.460, GD: 0.543), PG770은 가장 낮은 다형성을 나타내었다 (PIC: 0.070, GD: 0.078). 각 좌위들의 예상 이형접합도 (H$_{exp}$)는 0.077에서 0.541 (mean = 0.313)로 계산되었으나, 관측 이형접합도 (H$_{obs}$)는 0.040에서 0.130 (mean = 0.083)으로 훨씬 낮게 관찰되었으며, 유전자형의 분포는 Hardy-Weinberg 평형상태에서 벗어남을 보였다. 본 연구에서 개발한 인삼의 microsatellite 마커들은 인삼의 분자적 마커의 데이터베이스 확립의 기초 자료로 활용될 뿐 아니라, 인삼의 분자적 구별법 및 QTL 좌위의 염색체지도 작성에 유용하게 활용될 것이다.

• The Korean Journal of Physiology and Pharmacology
• /
• 제3권5호
• /
• pp.501-505
• /
• 1999

Tryptophan hydroxylase (TPH), the rate-limiting enzyme in serotonin biosynthesis, is primarily expressed in serotonergic neurons of the raphe nuclei. Simple tandem repeat polymorphisms, typically one to four nucleotides long, are tandemly repeated several times and often characterized by many alleles. To identify the presence of polymorphic repeats, we sequenced the 5'-upstream region of the mouse TPH gene. For the detection of any allelic variants, polymerase chain reaction, nonisotopic single-strand conformation polymophism, and DNA sequencing analyses of the tandem repeat sequences were performed using genomic DNA extracted from 60 ICR mice. Two dinucleotide repeats, $5'-(AC/TG)_{22}-3'$ and $5'-(GT/CA)_{17}3',$ were identified at approximately - 5.7 kb and - 3.4 kb upstream from the transcriptional initiation site of the mouse TPH gene, respectively. Minor allelic variants, $5'-(AC/TG)_{21}-3'$ and $5'-(GT/CA)_{18}-3',$ were observed in heterozygous pairs from 3 of 60 and 1 of 60 ICR mice, respectively. The identification of these microsatellites in the mouse TPH promoter raises the possibility that identical and/or other polymorphic sequences might exist in the upstream region of the human TPH gene.

• Animal cells and systems
• /
• 제16권1호
• /
• pp.50-56
• /
• 2012

Kelp grouper ($Epinephelus$ $bruneus$ Bloch 1793) is a commercially important fish in Korea. In recent years, the catch of kelp grouper in the coastal waters of Korea has significantly declined. Despite its importance, little is known about its genetic diversity and conservation efforts are hampered. In this study, we isolated and characterized 12 microsatellite loci using an enrichment method based on magnetic/biotin capture of microsatellite sequences from a size-selected genomic library. All loci were readily amplified and contained TG/CA denucleotide repeats. To characterize each locus, 30 individuals from a natural E. bruneus population in the coastal waters of Jeju Island, Korea, were genotyped. All loci except three, KEm118, KEm154, and KEm219, were polymorphic, with an average of 8.1 alleles per locus (range 2-18). The mean observed and expected heterozygosities were 0.47 (range 0.19-1.00) and 0.61 (range 0.29-0.92), respectively. A significant deviation from Hardy-Weinberg equilibrium was observed at three loci (KEm134, KEm184, and KEm283). These findings will be useful for effective monitoring and management of genetic variation of kelp grouper as well as for the implementation of a fisheries conservation program.

• Clinical and Experimental Reproductive Medicine
• /
• 제31권4호
• /
• pp.245-251
• /
• 2004

Objectives: To investigate the distribution and functional significance of $CYP11{\alpha}$ $(tttta)_n$ microsatellite polymorphism in Korean patients with polycystic ovary syndrome Materials and Methods: Analysis of $CYP11{\alpha}$ $(tttta)_n$ microsatellite polymorphism was carried out on DNA samples from 97 patients with polycystic ovary syndrome and 70 normal controls. Comparison were done between PCOS patients and controls concerning $CYP11{\alpha}$ $(tttta)_n$ microsatellite polymorphism genotype or allele frequencies. Results: The most frequent allele observed in the controls was an allele with six repeats (60.7%). Significant difference in the frequency of genotype (4R (-) genotype) having no copy of four-repeatallele were observed between PCOS patients and controls (66.0% vs 34.0%, p=0.038, OR=1.939). But no significant difference was observed in the serum levels of total testosterone or free testosterone between 4R (+) genotype and 4R (-) genotype among PCOS patients. However, hyperandrogenic PCOS patients with 4R (+) genotype showed a higher serum testosterone levels compared to controls (mean $\pm$ S.D: $0.49{\pm}0.21\;ng/ml$ vs $0.37{\pm}0.18\;ng/ml$, p=0.037). Conclusion: The alleleic distribution of $CYP11{\alpha}$ $(tttta)_n$ microsatellite polymorphism in Korean subjects were different from those reported in Caucasians. $CYP11{\alpha}$ $(tttta)_n$ microsatellite polymorphism was associated with polycystic ovary syndrome in the Korean population, and may play a role in the synthesis of androgens in patients with polycystic ovary syndrome.

• Tuberculosis and Respiratory Diseases
• /
• 제48권1호
• /
• pp.24-32
• /
• 2000

연구배경: MMR 유전자의 불활성화에 의해 야기되는 유전적 불안정성은 발암기전의 한 부류로 인정되고 있다. 저자들은 비소세포폐암의 발암과정에서의 MSI의 역할을 규명하기 위해 비소세포폐암에서 MSI의 빈도 및 MSI 유무에 따른 임상상의 차이를 조사하였다. 대상 및 방법: 근치적 절제술을 받은 비소세포폐암 20예를 대상으로 하였다. 동결된 폐암조직과 환자의 림프구에서 DNA를 추출한 후 3p와 9p의 15개의 marker들을 대상으로 PCR을 시행하고 7% polyacrylamide gel에서 전기영동한 후 silver 염색을 시행하였다. 암조직과 림프구 DNA의 PCR product의 band를 비교하여 MSI와 LOH를 판정하였다. 결 과: 1) 대상환자들은 남자 19예, 여자 1예였으며 모두 흡연자였고 평균 흡연력은 47 갑년이었다. 폐암의 조직형은 편평상피암 15예, 선암 4예, 대세포암 1예였고, 술후병리학적병기는 I기 6예, II기 5예, IIIA기 7예, IIIB 기 2예였다. 2) 20예 가운데 8예(40%)에서 MSI가 관찰되었으며 3예는 한 개의 marker에서, 5예는 2개 이상의 marker에서 MSI가 관찰되었다. 3) LOH는 10예(50%) 에서 있었으며, LOH 유무에 따른 병기 및 흡연력의 차이가 없었다. 4) 분석한 marker의 10% 이상에서 MSI가 관찰된 MSI-L 종양은 5 예였으며, 대부분의 marker에서 MSI 양성인 MSI-H 종양은 없었다. MSS 종양과 MSI-L 종양은 흡연력, 병기, 폐암 조직형 및 LOH 빈도의 유의한 차이가 없었다. 결 론: 비소세포폐암에서 MSI는 비교적 흔히 관찰되지만 MMR 유전자의 불활성화에 의한 MMP pathway는 비소세포폐암의 주요 발생기전은 아닐 것으로 생각된다. 향 후 비소세포폐암의 발암과정에 있어서 MMP pathway의 역할을 규명하기 위해서는 보다 많은 예를 대상으로 한 연구가 필요할 것으로 생각되며, MSI 발생기전에 관한 추가적인 연구가 필요할 것으로 생각된다.

• Asian-Australasian Journal of Animal Sciences
• /
• 제15권12호
• /
• pp.1822-1830
• /
• 2002

The microsatellites are the short tandem repeats of 1 to 6 bp long monomer sequences that are repeated several times. These short tandem repeats are considered to be generated by the slipped strand mispairing. Based on the unique capability of alternating purine-pyrimidine residues to form Z-DNA, the possible role of the microsatellites in gene regulation has been proposed. The microsatellites are highly polymorphic, follow Mendelian inheritance and are evenly distributed throughout the genomes of eukaryotes. They are easy to isolate and the polymerase chain reaction based typing of the alleles can be readily automated. These properties make them the preferred markers for comparison of the genetic structure of the closely related breeds/populations; very high-resolution genetic mapping and parentage testing etc. The microsatellites have rapidly replaced the restriction fragment length polymorphism (RFLP) and the random amplified polymorphic DNA (RAPD) in most applications in the population genetics studies in most species, including the various farm animals viz. cattle, buffalo, goat, sheep and pigs etc. More and more reports are now available describing the use of microsatellites in pigs ranging from measurement of genetic variation between breeds/populations, developing high resolution genetic maps to identifying and mapping genes of biological and economic importance.

• Biotechnology and Bioprocess Engineering:BBE
• /
• 제5권1호
• /
• pp.23-26
• /
• 2000

In order to develop the specific genetic marker for Korean native cattle (Hanwoo), randomly amplified polymorphic DNA (RAPD) analysis of 6 different cattle breeds was attempted by using 38 decamer primers. In comparison of RAPD patterns, two distinctive DNA bands specific for Hanwoo were detected. One was 296 bp of DNA fragment found to be specific only for female Hanwoo when primer GTCCACACGG was employed. In individual analysis of this RAPD marker was observed only in female individuals with the possibility of $85.3\%$. The other was 521 bp of RAPD marker amplified using TCGGCGATAG and AGCCAGCGAA primers, which showed $83.0\%$ of genetic frequency in 85 male and 68 female individuals tested. Nucleotide sequencing of these genetic markers revealed that 296 bp marker has a short micro satellite-like sequence, ACCACCACAC, and a tandem repeat sequence of microsatellite GAAAAATG in the determined sequence. Two distinctive tandem repeats of microsatellite sequences, MC and GAAGA, were also appeared in 521 bp DNA marker. In BLAST search, any gene having high homology with these markers was not found.

• Fisheries and Aquatic Sciences
• /
• 제14권4호
• /
• pp.303-310
• /
• 2011

We characterized a cytoplasmic actin gene locus in greenling Hexagrammos otakii (Scorpaeniformes). Genomic clones isolated from the greenling DNA library contained two homologous cytoplasmic actin gene copies (HObact2.1 and HObact2.2) in a tail-to-head orientation. Their gene structure is characterized by six translated exons and one non-translated exon. Exon-intron organization and the nucleotide sequences of the two actin gene isoforms are very similar. However, only the HObact2.1 isoform contains microsatellite-like, dinucleotide repeats in the 5'-flanking region (named HOms2002) and intron 1 following the non-translated exon 1 (named HOms769). One microsatellite locus (HOms769) was highly polymorphic while the other (HOms2002) was not. Based on bioinformatic analysis, different transcription factor binding motifs are related to stress and immune responses in the two actin isoforms. Semiquantitative and real-time reverse transcription-PCR assays showed that both isoform transcripts were detectable ubiquitously in all the tissues examined. However, the basal expression levels of each isoform varied across tissues. Overall, the two isoforms showed a similar, but not identical, expression pattern. Our data suggest that the cytoplasmic actin genes may be the result of a recent duplication event in the greenling genome, which has not experienced significant subfunctionalization in their housekeeping roles.

• Asian-Australasian Journal of Animal Sciences
• /
• 제31권2호
• /
• pp.167-179
• /
• 2018

Objective: The aim of this study was to create a set of microsatellite markers with high polymorphism for the genetic monitoring and genetic structure analysis of local goose populations. Methods: Novel microsatellite markers were isolated from the genomic DNA of white Roman geese using short tandem repeated probes. The DNA segments, including short tandem repeats, were tested for their variability among four populations of geese from the Changhua Animal Propagation Station (CAPS). The selected microsatellite markers could then be used to monitor genetic variability and study the genetic structures of geese from local geese farms. Results: 14 novel microsatellite loci were isolated. In addition to seven known loci, two multiplex sets were constructed for the detection of genetic variations in geese populations. The average of allele number, the effective number of alleles, the observed heterozygosity, the expected heterozygosity, and the polymorphism information content were 11.09, 5.145, 0.499, 0.745, and 0.705, respectively. The results of analysis of molecular variance and principal component analysis indicated a contracting white Roman cluster and a spreading Chinese cluster. In white Roman populations, the CAPS populations were depleted to roughly two clusters when K was set equal to 6 in the Bayesian cluster analysis. The founders of private farm populations had a similar genetic structure. Among the Chinese geese populations, the CAPS populations and private populations represented different clads of the phylogenetic tree and individuals from the private populations had uneven genetic characteristics according to various analyses. Conclusion: Based on this study's analyses, we suggest that the CAPS should institute a proper breeding strategy for white Roman geese to avoid further clustering. In addition, for preservation and stable quality, the Chinese geese in the CAPS and the aforementioned proper breeding scheme should be introduced to geese breeders.

• 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.