• Journal of Ginseng Research
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• v.14 no.2
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• pp.310-316
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• 1990

This study was focused on the characterization of mitochondrial DNA (mtDNA) for molecular 9enetical approach of energy Production related mechanism in Panax ginseng. The simple and efficient method of mtDNA isolation from ginseng has been developed by modification of recently advanced methods. This procedure can successfully apply to mtDNA isolation of several plants. mtDNA of etiolated shoot and one-year root were digested with restriction endonucleases, but that of 6-year root not. Any difference was not observed in the restriction endonuclease digestion patterns among the ginseng variants. Molecular size of ginseng mtDNA was estimated at least 159 kb by the restriction endonuclease fragment analysis. The 4.5 kb extra band at the lane of EcoRII treatment could be observed in restriction patterns digested with the methylation sensitive endonucleases, BstN I and EcoRII. For construction of mitochondrial genomic library of ginseng, mtDNA was partially digested with EcoRl, and packaged with EMBL4 phage vector. Genomic library was screened and purified for further research including restriction mapping of ginseng mtDNA, and cloning of the genes. The gene of ATP synthase A subunit was cloned from the purified EMBL4 library clone No. 16. Now, clone No. 16 is subcloned for structure gene sequence analysis.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.30 no.5
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• pp.804-808
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• 1997

The genetic differentiation and characteristics of two oyster populations (Crassostrea gigas) in Korea were assessed based on the restriction fragment length polymorphisms (RFLP) analysis and the restriction patterns of subcloned mtDNA. The restriction fragments of twenty individuals in West Sea revealed an identical pattern, determined by 8 restriction enzymes. On the other hand, two haplotypes having variation at the HindIII site were shown in the specimens from South Sea; minor haplotypes (4 of 20) were similar to the results obtained from individuals in West Sea while major haplotypes were different from those in West Sea. It was suggested that oysters (C. gigas) of West Sea might have been introduced to South Sea. Each mitochondrial DNA from two oyster populations in Korea and from one in Japan was divided to three parts and subcloned into pUC19 to use in genetic studies effectively. Restriction map was constructed based on the cleavage pattern by multiple restriction enzymes.

• The Journal of the Korean Society for Microbiology
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• v.22 no.4
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• pp.463-471
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• 1987

The genomes of leptospiral field isolates from Korea belonging to serogroup Icterohaemorrhagiae (21 strains) and serogroup Canicola (1 strain) were analysed and compared by restriction enzyme analysis with EcoRI and HindIII as digesting enzymes. One isolate belonging to serogroup Canicola showed the same pattern as serovar portlandvere. All 21 isolates belonging to serogroup Icterohaemorrhagiae showed almost same patterns as Leptospira serovar lai from China, But with very slight differences 21 isolates could be classified into 8 subtypes and these grouping seems to reflect the differences in epidemiological niche. And also the geographical data consisted with the grouping into 8 subtypes. According to our results, we concluded that the restriction endonuclease analysis of chromosomal DNA will be an accurate and reliable method to compare and classify pathogenic leptospires.

• Proceedings of the Ginseng society Conference
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• 1990.06a
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• pp.168-174
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• 1990

This study was focused on the characterization of mitochondrial DNA (mtDNA) for molecular genetically approach of energy Production related mechanism in Panax Ein.fend. The simple and efficient method of mtDNA isolation from ginseng has been developed by modification of recently advanced methods. This procedure can successfully apply to mtDNA isolation of several plants. MtDNA of etiolated shoot and one-year root were digested with restriction endonucleases, but that of 6-year root not Any difference was not observed in the restriction endonuclease digestion patterns among the ginseng variants. Molecular size of ginseng mtDNA was estimated at least 159 kb by the restriction endonuclease fragment analysis. The 4.5 kb extra band at the lane of EcoRll treatment could be observed in restriction patterns digested with the methylation sensitive endonucleases, BstN 1 and EcoRll. For construction of mitochondrial genomic library of ginseng, mtDNA was partially digested with EcoRl, and packaged with EMBL4 phage vector Genomic library was screened and purified for further research including restricttion mapping of ginseng mtDNA, and cloning of the genes. The gene of ATP synthase A subunit was cloned koto the purified EMBL4 library clone No. 16. Now, clone No. 16 is subcloned for structure gene sequence analysis.

• Restorative Dentistry and Endodontics
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• v.20 no.2
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• pp.577-609
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• 1995

Bacteria have been regarded as one of the most important factors in pulpal and periapical diseases. Streptococci are frequently isolated facultative anaerobes in infected root canals. Recently molecular biological techniques have been rapidly progressed. This study was designed to apply the molecular biological tools to the identification and classification of streptococci in the endodontic microbiology. Streptococci isolated from infected root canals were identified with both Vitek Systems and API 20 STREP. Identification results were somewhat different in several strains of streptococci. Eighteen streptococci and enterococcal was difficult so to digest plasmid DNA using Hind III and EcoRI to differentiate strains by restriction enzyme analysis of plasmid DNA. 16S rDNA of chromosome was amplified by polymerase chain reaction(PCR) and then restricition fragment length polymorphism(RFLP) using several restriction enzymes was observed. The molecular mass of 16S rDNA of chromosomal DNA was approximately 1.4kb. There were three to five RFLP patterns using eight restriction enzymes. RFLP patterns digested with CfoI which recognizes four base sequences were identical in all stains. Hind III which recognizes six base sequences could not digest the 16S rDNA. Restriction enzymes which recognize five base sequences were suitable for RFLP pattern analysis. At least three different restriction enzymes were needed to compare each strains. 16S rDNA PCR-RFLP was simple and rapid to differentiate and classify strains and could be used in the epidemiological study of root canal infections.

• Korean Journal of Environmental Biology
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• v.18 no.1
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• pp.53-61
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• 2000

In order to characterize the genetic diversity of bacterial community in groundwater, samples were collected from used for drinking water and polluted with heavy metal wastewater in Seoul city and natural cave of Kangwondo. The DNA was amplified with 165 rDNA-based primers by use of the PCR, and then analysed ARDRA (amplified ribosomal DNA restriction analysis). Restriction endonuclease analysis patterns of amplified 165 rDNA in drinking water and wastewater relatively showed high genetic diversity in situ and drinking groundwater. The number of DNA fragments varied with in situ and drinking water. This method of ARDRA of bacterial communities in groundwater could be used for a quick assessment of genotypic changes between different locations reflecting different environmental conditions and the diversity reflected pollution of groundwater (natural cave water>drinking water>waste water, as in order of grade). [Genetic diversity, Groundwater, 165 rDNA, PCR, ARDRA].

• Korean Journal of Veterinary Research
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• v.33 no.1
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• pp.43-51
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• 1993

Mitochondrial DNA(mt DNA) of Mammalian is the circular one which the 16.5K base pairs and show the maternal inheritance. Evolutional speed of nucleotide sequence is very fast. So that polymorphic analysis of mt DNA provide the useful informations to investigate the genetic relations of interspecies. Authors trials were focussed to compare with the polymorphic differences of mitochondrial DNA between Jindo and Japanese mongrel dogs. DNA was extracted from bloods of 21 head of Jindo dogs and 20 head of Japanese dogs and isolated using 10 kinds of restriction endonucleases(Apa I, BamH I, Bgl II, EcoR I, EcoR V, Hinc II. Hind III, Pst I, Sty I, Xba I) and then separated by the agarose gel electrophoresis. After sourthern blotting hybridization was completed using the mtDNA of Japanese mongrel dogs as a probe. Autoradiography was used to compare the polymorphism of mtDNA both dogs. The results obtained were as follows; 1. mt DNA of Jindo dog showed polymorphism resulting cleavage with four kinds of restriction endonuclease, Apa I, EcoR V, Hinc II, Sty I. While in the Japanese mongrel dogs observed the polymorphism in the five kinds of restriction endonuclease supplemented with EcoR I. 2. Compared with both dogs the frequency differences of DNA polymorphism were recognized in the specific restriction endonuclease Apa I. Consequently in the restriction endonuclease Apa I both dogs classified with three types as A, B, C however in the Jindo dogs frequency of C type was 71.5 percent but in Japanese mongrel dogs observed 45 percent in the A type. 3. DNA polymorphism obtained from the use of five kinds of restriction endonuclease were classified with seven types. In Jindo dogs frequency was highest in the type 6 as 71.4 percent but in the Japanese mongrel dogs showed 35 percent in the type 5. 4. Genetic distances calculated by NEI method showed 0.0089 in Jindo dogs and was 0.0094 in the Japanese mongrel dogs.

• Korean Journal of Microbiology
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• v.23 no.2
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• pp.115-121
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• 1985

Five representative virulent phages (J1, TK93, K1, PD5, and CP1) and one temperate phage (.phi.1043) of Lactobacillus casei were compared to each other by analyzing the agarose gel electrophoretic patterns of restriction enzyme-digested phage DNAs. Nucleic acids of all the tested phages were double stranded DNA. DNAs of J1, TK93, K1, and ${\phi}$ 1043 phages had a size of about 42kb, but the size of PD5 and CP1 DNAs was avout 140kb. J1, TK93, K1, PD5, CP1, and ${\phi}$ 1043 DNAs were digested to 13, 13, 11, 14, 14, and 12 fragments by EcoR1, respectively, and showed its characteristec restriction patterns. Cohesive ends were present in J1, TK93, and ${\phi}$ 1043, but were absent in K1, PD5, and CP1. Restriction maps of J1 and TK93 DNAs showed nearly complete homology and their evolutionary relationship based upon the restriction analysis was discussed.

• Animal cells and systems
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• v.3 no.1
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• pp.73-78
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• 1999

Although there are several methods for the preparation of mitochondrial DNA (mtDNA) from mammalian tissues, most are relatively long ultracentrifugation or manipulations by a small-scale method. We escribed a rapid method for large-scale extraction of mtDNA from human placental and horse liver tissues. The method is based on the preparation and homogenization of tissues, urification of crude mitochondria by differential centrifugations and isolation of mtDNA by alkaline Iysis. It was improved from Pre-existing methods by replacing some steps with simpler ones and discarding many others. This method gives a high yield of pure mtDNA(approximately 1-5mg from one placenta; ca. 400-600 g wet weight), depending on its sources (fresh tissue gave better results than frozen one). The resulting mtDNA indicated that this method can yield mtDNA in sufficient purity and quantity to identify the direct restriction analysis on agarose gel, random-primed labeling as a probe, and end labeling. Therefore, the method is ideal for obtaining good mtDNA samples to conduct routine restriction fragment length polymorphism (RFLP) analyses of natural populations for genetic studies.

• 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.