• Korean Journal of Fisheries and Aquatic Sciences
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• v.27 no.5
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• pp.613-619
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• 1994

To investigate the population differences of small yellow croaker (Pseudosciaena polyactis BLEEKER) in the Yellow Sea, five catching sites (three from China; Zoushan, Shanghai and Qingdao, two from Korea; Inchon and Mokpo) were selected for sampling. The populations of small yellow croaker from all five catching sites were investigated to analyze their mtDNA's restriction fragment length polymorphism (RFLP) using 18 kinds of restriction enzymes. The average molecular size of the entire mtDNA was estimated at $16.9{\pm}0.6\;kb$. According to the results of RFLP analysis, a total of 40 restriction sites were identified in every population surveyed and the overall cleavage patterns of mtDNA, based on the RFLP, showed similar tendencies. However, the five restriction enzymes such as ApaI, EcoRI, PstI, SmaI and SstII showed slightly different cleavage patterns which could have resulted from individual variations between the populations of Korea and China.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.30 no.1
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• pp.88-94
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• 1997

The mitochondrial DNA (mtDNA) restriction fragment length polymorphi는 (RFLPs) of five populations were analyzed to delineate the stocks of Penaeus chinensis (Osbeck) in the Yellow Sea. Comparison of P. chinensis with P. japonicus to clarify the nucleotide divergence between two species was also carried out. Based on the fragment patterns, three composite haplotypes were analyzed in P. chinensis mtDNA as four naplotypes were in P. japonicus. Most individuals of each P. chinensis population are shared by one dominant haplotype. Another two haplotypes haying variations at the C/a I and hull sites were also distributed evenly in the Korean and Chinese populations. It is suggested that the gene exchange occurring between populations in the Yellow Sea is frequent. Average length of the mtDNA molecule was estimated to be about 16.44 kb in P. chinensis and 16.31 kb in P. japonicus, Sequence divergence (p) of mtDNA between two species estimated by using Upholt's (1977) fomula was $13.7\%$.

• The Korean Journal of Mycology
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• v.28 no.2
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• pp.112-117
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• 2000

A similarity coefficient were analyzed by RFLP of fourteen species of entomopathogenic fungi, isolated from inhabiting pupa and adult insect at forest. Each rDNA ITS I and ITS II with primers of ITS 1 and ITS 4 was amplified by PCR. The amplified products were conserved to 500 bp were not demarcated between genus and species. Four Paeciliomyces tenuipes, two Beauveria bassiana and six Cordyceps militaris were treated by seven restriction enzymes and confirmed in species except JB3 by electrophoresis band. However, the band of C. scarabaeicola showed the identity with B. bassiana. The result of this experiment indicated that the teleomorph of C. scarabaeicola was the same as that of B. bassiana. CfoI and HpaII of seven restricted enzymes were easily discriminating in the genus between Paecilomyees and Cordyceps. Especially, CfoI was more effective to classify the genera of Paecilomyees, Cordyceps and Beauveria than other restriction enzymes. The band patterns of RFLP of P. tenuipes, C. militaris, C. scarabaeicola and B. bassiana were also analyzed by UPGMA program of NTSYS-pc and showed 100% significance. Thus, the similarity coefficient tended to be lower between genera by RFLP analysis, but was higher between species.

• Parasites, Hosts and Diseases
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• v.35 no.2
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• pp.119-126
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• 1997

Acanthnmoebn sp. YM-4 is simitar to A. culbertsoni based upon morphological characteristics of trophozoites and cysts. However, based on other characteristics, pathogenicity to mice, in uitro cytotoxicity and isoenzyme patterns, Acanthomoebo sp. YM- 4 was quite different from A. culbertsoni. Restriction fragment length polymorphism (RFLP) analysis of mtDNA is useful in the classification of members belonging to the genus Acanthcmoebn. Therefore, in this study, RFLP analysis of Acnnthcmoeba mtDNAs was accomplished using five restriction enzymes: Hnelll, Hinull, Clcl, Pudl and ScE. Each restriction enzyme produced approximately 3-15 fragments (range: from 0:6 kip to 34.4 kbp) . The mtDNA genome size, calculated by the summation of restriction fragments, averaged 46.4 kbp in Acnnthamoeba sp. YM-4,48.3 kbp in A. culbertsoni and 48.8 kbp in A. polyphaic, respectively. Digested mtDNA fragments of Accnthcmoeba sp. YM-4 contained nine and seven same size fragments, respectively, from a total of 67 and 69 fragments observed in A. culbertsoni and A. polyphcgn. An estimate of the genetic divergence was 10.1% between Acanthamoebc sp. YM-4 and A. culbertsoni, and 9.9% between Acanthamoebn sp. YM-4 and A. polyphcga.

• Korean Journal of Microbiology
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• v.32 no.4
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• pp.245-251
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• 1994

Ten strains of 7 species from the genus Ganoderma, G. lucidum ATCC 64251, FP-103561-T, and ES70701, G. applanatum ATCC 44053 and FP-57035-T. G. lobatum ATCC 42985, G. resinaceum ATCC 52416, G. subamboinense var. laevisporum ATCC 52420, G. meredithae ATCC 64492, and G. microsporum ATCC 76024, were studied to discuss their phylogenetic relationships by utilizing restriction fragment length polymorphisms (RFLPs) of mitochondrial DNAs (mtDNAs). Six restriction enzymes, BamHI, BglII, EcoRI, HindIII, PvuII, and XbaI which digested mtDNAs into adequate numbers of restriction fragments for cluster analysis, were used in this study. Restriction profiles of strains for each restriction enzyme were treated as analysis characters to calculate similarity coefficients, which were converted into nucleotide sequence divergence values whose mean values were then arranged in a matrix table. This table was utilized for a phylogenetic analysis using the Neighborjoining method of the PHYLIP package to construct phylogenetic tree. Three strains of G. lucidum and two strains of G. applanatum exhibited different lineages each but one of G. applanatum strains showed a close relationship with G. lobatum, which reflected the species complexity of these species whose strains were phenotypically indistinguishable but genetically distinct. The present results suggest that the natural classification of Ganoderma needs to be considered from the viewpoints of molecular biology-based systematics as well as morphological classifications and cultural identifications for better phylogenetic conclusions.

• 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.

• Journal of Science Education
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• v.46 no.3
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• pp.293-311
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• 2022

In this study, a restriction enzyme mapping inquiry experiment was developed for cultivating basic knowledge on molecular biology and the effects on inquiry experiment ability and positive experience on science through student-centered molecular biology inquiry experiment class for second graders of a general high school was analyzed. First of all, it was found that the experimental class through the inquiry experiment was significantly effective as the percentage of high school students who answered 'yes' or higher in the positive science experience of general high school students was higher after than before the test. As a result of developing and applying a series of five classes for the creation of restriction enzyme maps, not only did the students' interest in science studies, but also their class participation increased. They were also used as effective specific science learning motives, science career aspirations and experience data. The science environment of the inquiry experiment class led to the improvement of students' learning attitudes and positive science experience, which had a positive effect on the importance of class concentration and class quality, active communication and mutual cooperation among students. In addition, inquiry and experiment classes will provide opportunities for career experience, which will become the foundation for cultivating basic knowledge on molecular biology and advancing to science and engineering.

• Korean Journal Plant Pathology
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• v.12 no.2
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• pp.169-175
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• 1996

콩 불마름병균 Xanthomonas campestris pv. glycines 8ra는 X. c. pv. vesicatoria에 길항력이 있는 bacteriocin인 glycinecin을 생성 분비한다. Bacteriocin 생성 분비 능력이 있는 콩 불마름병균을 효과적인 생물학적 방제원으로 활용하기 위해서는 좀더 체계적인 연구가 필요하여, bacteriocin 생성에 관계되는 유전자의 분리를 시도하였다. 약 2,000개의 Xanthomonas campestris pv. glycines 8ra cosmid library에서 bacteriocin의 생성 분비 능력을 조사하여 다섯 개의 clone을, pG011, pG0113, pG33과 pG35, 선발하였다. 그중 한 clone pG08을 임의로 선택하여 plasmid DNA를 분리하였다. Plasmid pG08에서 약 6.0 kb의 DNA를 떼어내어 다른 plasmid vector에 넣은 subclone pBL5는 bacteriocin의 생성 분비 능력이 있었다. Plasmid pG08을 제한효소 처리후 다시 접함시켜 만든 몇 개의 subclone과 pBL5의 제한효소 지도를 비교 분석한 결과 약 3.0 kb의 BamHI-HindIII 부분의 DNA가 bacteriocin의 생성에 관계함을 알았다.

• The Korean Journal of Malacology
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• v.13 no.1
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• pp.1-7
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• 1997

한국산 가리비, 큰가리비(Patinopecten yessoensis)와 주문진가리비(Chlamys swifti), 2종에 대한 28S ribosomal RNA 유전자의 PCR- 산물을 이용 RFLP 및 염기서열을 밝히고, 이미 보고된 2과 3종의 염기서열과 상동성을 비교 분석하였다. 그 결과 28S rRNA유전자를 이용하여 7가지 제한효소를 처리한 PCR-RFLP의 종간 차이에서 Taq I 제한효소에서만 차이를 볼 수 있었다. 한편 두종간에 28S rRNA유전자의 D1 부위의 염기서열에서 231개 부위 중 14군데에서 변이를 보였다.

• Biomedical Science Letters
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• v.6 no.1
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• pp.73-82
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• 2000

Blunt-end DNA fragments can be inserted in two different orientations. Conventionally, their directions are determined by restriction enzyme digestion or by DNA sequencing, however, these methods are often limited in their use due to the lack of appropriate enzyme sites or large sample numbers, respectively. In the present study, a novel strategy and the corresponding protocol for the simple determination of insert orientation is introduced. Using conventional sequencing primers and PCR primers that have been used for amplification of the insert, single clones, which have inserted the fragment in the desired orientation, were easily identified by this PCR-based method. The fidelity of this system was confirmed by cloning of a tar urocortin cDNA, which is a recently discovered neuropeptide. Recombinant clones identified by this method were further shown to be fully functional, and using these, for the first time, urocortin was recombinantly expressed in eukaryotic cells.