• Korean Journal of Environmental Biology
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• v.36 no.3
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• pp.352-358
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• 2018

This study aimed to develop a species identification method for the egg and fry of the three Korean bitterling fishes (Pisces: Acheilognathinae), including Acheilognathus signifer, Acheilognathus yamatsutae and Rhodeus uyekii based on the PCR-based Restriction Fragment Length Polymorphism (RFLP) markers. We conducted a field survey on the Deokchicheon River from the North Han River basin, where the three Acheilognathinae species co-occur, and also analyzed the existing sequence dataset available from the GenBank. We found coexistence of the three species at the study site. The egg and fry were obtained from the host mussels (Unio douglasiae sinuolatus) by hand from May to June 2015 and in May 2017. To develop PCR-based RFLP markers for species identification of the three Acheilognathinae fish species, restriction enzymes pinpointing species-specific single nucleotide variation (SNV) sites in mitochondrial DNA COI (cytochrome oxidase I) and cyt b (cytochrome b) genes were determined. Genomic DNA was extracted from the egg and fry and RFLP experiments were carried out using restriction enzymes Apal I, Stu I and EcoR V for A. signifer, A. yamatsutae and R. uyekii, respectively. Consequently, unambiguous discrimination of the three species was possible, as could be seen in DNA band patterns from gel electrophoresis. Our developed PCR-based RFLP markers will be useful for the determination of the three species for the young and would assist in studying the spawning patterns and reproductive ecology of Acheilognathinae fishes. Furthermore, we believe the obtained information will be of importance for future maintenance, management and conservation of these natural and endangered species.

• Journal of Life Science
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• v.15 no.6 s.73
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• pp.879-883
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• 2005

This study was differentiated between Korea and China arkshells using PCR-aided RFLP method which could identify the variation for inter-and intra-species of arkshell (Scapharca broughtonii Schrenck) at the level of DNA. The DNA fragment patterns were compared after digesting gene of mitochondrial 16S rDNA with 8 kinds of restriction enzymes. A 720 bp DNA fragment corresponding to 16S rDNA gene was amplified by PCR with primers ArkF-3 and ArkR-3. PCR products were cut by restriction enzymes (Pvull, BamHI, Hinfl, HaeIII, EcoRI, RsaI, Ksp221, and BstX21), and RFLP pattern was studied. A unique 275 bp DNA band was observed in the samples from Dukyang, Gamak, Namhae, Jinhae, and Taean in Korea when treated by Hinfl, but Chinese arkshell did not show. Treatment of HaeIII could discriminate the sample of Namhae and Jinhae from Dukyang/Gamak/Taean, as well as Korean and Chinese arkshell based on a 700 bp. However, PuvII, BamHI, EcoRI, RsaI, Ksp221, and BstX21 showed the same of 700 bp band in Korean and Chinese arkshell. The phylogenetic tree inferred from PCR-RFLP pattern comparsion in Korean arkshell was different that the distance between Dukyang/Gamak/Taean and Namhae/Jinhae was approximately 7. In particular, the distance between Korean and Chinese arkshell was 25. Consequently, HinfI and HaeIII played an important role in a reliable molecular tool for rapid discriminating Korean and Chinese arkshell, as well as a intra-species in Korea.

• The Korean Journal of Malacology
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• v.13 no.2
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• pp.91-96
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• 1997

한국산 논우렁이(CIpangopaludina chinensis malleata)와 큰논우렁이 (C. japomica)는 형태학적으로 유사하여 그 감별이 용이치 않다. 본 연구는 이 두 종을 대상으로 28S rDNA DI유전자를 7종의 제한효소로 처리하여 PCR-RDLP기법으로 그 절편을 비교하였다. 절편 상호간에는 차이점을 관찰할 수 없었으나, 두 종으로부터 분석된 28S rDNA DI 유전자의 염기서열에서는 4 부위에서 종간 차이를 보였다.

• Korean journal of applied entomology
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• v.37 no.1
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• pp.23-30
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• 1998

Restriction fragment length polymorphism (RFLP) of a DNA has been a useful tool for analyzing genetic variation. This research was performed to establish an RFLP analytic method on the mitochondrial DNA (mtDNA) of the beet armyworm, Spodoptera exigua (Hiibner). To do this, total size of the mtDNA was measured and polymerase chain reaction (PCR) primers were selected. Its mitochondrial genome size was ca. 16kb. From a serial PCR test of 29 primers refered to the compilation of Simon et al. (1994), 22 primers were selected to amplify its mtDNA fragments. These primers resulted in short (300-700 bp) or long (1000-2000 bp) DNA products which represented a total or partial sequence of each of CO-I, CO-11, Cyt-B, ND-1, 12s rRNA, 16s rRNA, and some tRNAs. PCR-RFLP was performed in some variable mtDNA regions with 8 kinds of 4bp recognizing restriction enzymes. Different populations from Andong, Kyungsan, and Sunchun did not show any restriction site polymorphisms but had some length variation in certain regions of mtDNA.

• Korean Journal of Biological Psychiatry
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• v.2 no.2
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• pp.248-251
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• 1995

정신분열병의 약물치료에서 중요한 역할을 하는 도파민의 기전은 지금까지 5종류의 수용체들이 발견되고 클론되면서, 새롭고 보다 근본적인 유전학적 접근을 통해 규명될 수 있게 되었다. 특히, 도파민 수용체 D4 (DRD4)는 막단백질의 세포질쪽 세번째굴곡에 48-bp반복 다형성배열을 가지고있다. 이러한 다형성 반복배열이 신호전달에 참여할 가능성이 높은 막단백질의 세포질쪽 굴곡에 있다는것은, 각 개인의 항정신병약물에 대한 민감성의 차이를 포함하여 정신분열병에 대한 개개인의 유전적 차이를 진단할 수 있는 가능성을 제시한다. 이러한 가능성을 검증하기 위하여 주로 손쉽고 빠른 중합효소연쇄반응(PCR)을 사용하여 DRD4의 아형들을 분류하게 되는데, DRD4의 PCR은 그 반복배열과 그 주변배열의 높은 GC함량(78% G+C) 때문에 일반적인 PCR 방법을 변형시켜 사용해야한다. DRD4의 아형을 분류하기 위해 변형된 PCR은 통상적으로 7-deaza dGTP와 10% DMSO를 사용하게된다. 이러한 DRD4 PCR은 대부분의 경우 성공하지만, 항상 모든 시료에서 PCR이 성공되는것은 아니었으며 반복적으로 시도하여 증폭시킬 수 있었다. 이러한 어려움은 대부분이 template DNA에 문제가 있을것으로 의심되며 DNA정제 또는 template DNA를 제한효소로 적절하게 무작위절단하여 성공율을 높일 수 있었다.

• Journal of Periodontal and Implant Science
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• v.29 no.2
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• pp.349-355
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• 1999

The present study has been performed to develop a PCR technology to identify human immunoglobulin(Ig) allotypes with restriction fragment length polymorphism(RFLP) using a probe. Genomic DNA were ampilified with PCR tecnology using primers from peripheral blood lymphocytes of 10 periodontal patiens, whose Ig allotypes have been pre-determined by serological tecnique using heagglutination technique. The result indicated that the RFLP patterns could successfully differentiate the Ig allotypes, which suggests that this technology can be developed as a tool useful for population genetics studies.

• The Korean Journal of Mycology
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• v.27 no.1 s.88
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• pp.20-26
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• 1999

The specific DNA band appeared in PCR-RAPD analysis using OPO-2 primer was a very important for the researching Korean pine-mushrooms, Tricholoma matsutake. This DNA band, sequenced to be the 770 base pairs, existed as only a single copy in the whole genomic DNA's of Korean pine-mushrooms. However, this band was not presenting from the PCR-RAPD bands of other ectomycorrhyzal fungi reacted with the OPO-2 primer or the dot blots. Also, this DNA sequence was not matched with those of the other genes known by NCBI and had low homology together with sequence of other proteins compared. Those results suggested that the specific DNA band can be used as probe for identification of T. matsutake and might be related to the informations rather than the gene for the proteins with analysis of protein sequence translated from the DNA sequence.

• Journal of Food Hygiene and Safety
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• v.23 no.4
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• pp.314-318
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• 2008

This research aimed to compare the detection methods of Anisakis simplex in Sea fish by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and macroscopic inspection. We examined 18 Trichiurus lepturus, 11 Scomber japonicus, and 65 Todarodes pacificus collected from the retail markets in the areas of Uljin, Kyuonggi province and Seoul. As the result of examinations, we found that detection rate of Anisakis simplex by macroscopic observation was 89% in Trichiurus lepturus, 90.9% in Scomber japonicus, 32.3% in Todarodes pacificus. The detection rate of Anisakis simplex by PCR-RFLP was 77.7% in Trichiurus lepturus, 81.8% in Scomber japonicus, 26.1% in Todarodes pacificus. We could conclude that PCR-RFLP method of Anisakis simplex was more specific rather than macroscopic observation.

• Korean Journal of Plant Taxonomy
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• v.41 no.2
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• pp.119-129
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• 2011

Phylogenetic studies were conducted to evaluate the taxonomic relationships among 27 taxa, including 2 outgroups of Korean Campanulaceae, using PCR-RFLP analysis and ITS sequences. In the PCR-RFLP analysis, 15 restriction endonucleases produced 244 restriction sites and size variations from the chloroplast DNA, and 59 restriction sites (24%) showed polymorphism. The length of the ITS regions ranged from 588 bp to 797 bp. The sequence divergence including the outgroups is 0-39.36%. Phylogenetic analyses based on PCR-RFLP and ITS data suggest that Campanulaceae is monophyletic; Codonopsis and Platycodon forms an independent clade; the Peracarpa and Asyneuma clade is a sister to the Adenophora-Hanabusaya clade; Campanula is monophyletic; and Wahlenbergia basally branches within the ITS tree, whereas they are placed between Campanula and the Codonopsis-Platycodon clade in the PCR-RFLP tree; Hanabusaya is placed within the Adenophora clade; and Adenophora is paraphyletic and shows discordance to the infrageneric classifications based on morphological data. The present results show two data sets, largely congruent at the generic level, but their phylogenetic positions, in particular the Wahlenbergia and Hanabusaya and the infrageneric classifications in Adenophora, show some incongruence.

• Applied Biological Chemistry
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• v.40 no.6
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• pp.495-500
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• 1997

To isolate a gene for cyclodextrin glycosyltransferase (CGTase) from alkalophilic Bacillus sp. E1, polymerase chain reaction (PCR) amplification was carried out. Direct molecular cloning of 1.2 kbp fragment and partial nucleotide sequence analysis of the PCR amplified clone, pH12, showed close homology with CGTases from Bacillus species. To investigate the genomic structure of the gene, Southern blot analysis of genomic DNA was carried out with the clone pH12 as a molecular probe. It showed that 5.3 kbp XbaI fragment was hybridized with the probe pH12. To isolate a genomic clone, genomic DNA library was constructed and a genomic clone for CGTase, pCGTE1, was isolated. Nucleotide sequence analysis of the clone pCGTE1 revealed that BCGTE1 contained 2,109 bp open reading frame encoding a polypeptide of 703 amino acids and showed over 94.3% amino acid sequence homology with CGTase of ${\beta}-cyclodextrin$ producer, Bacillus sp. KC201.(Received October 7, 1997; accepted October 20, 1997)