• Korean Journal of Microbiology
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• v.36 no.4
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• pp.249-253
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• 2000

The genetic diversity among Korean cyanobacteria was assessed by restriction fragment length polymorphism(RFLP) analysis of PCR products from the phycocyanin locus. Strains of all the genera tested were successfully amplified, and the size of amplified fragments was approximately 700bp. The restriction patterns generated by AluI, MspI, and HaeIII were conserved for strains within each of genera studied and were specific to the genus level. Intrageneric delineation of strains was revealed by the enzyme, CfoI for members of genera Anabeana and Synechocystis. Phenogram derived from the different RFLP patterns revealed a coherent cluster among Anabeana, Chlorogloea, and Synechocystis strains. PC-RFLP methods provided useful tools for classification of cyanobacteria.

• Journal of Microbiology and Biotechnology
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• v.9 no.3
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• pp.334-339
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• 1999

More innovative molecular markers were investigated for rapid and consistent differentiation of Metarhizium anisopliae var. majus from M. anisopliae var. anisopliae. A total of 28 isolates were obtained from various countries and hosts: 13 isolates of M. anisopliae var. anisopliae, 12 isolates of M. anisopliae var. majus, and 3 isolates of M. anisopliae collected in Korea. This study involved restriction enzyme digestions of a PCR product amplified from nuclear internally transcribed spacer (ITS) and a portion of the 28S rDNA regions. Among 11 different restriction enzymes used in this study, MboⅠ digestion particularly produced a restriction pattern that had characteristics of M. anisopliae var. majus. This restriction pattern was consistent in all isolates of M. anisopliae var. majus regardless of their geographic origins and insect hosts. Mapping experiments revealed that MboⅠ sites of M. anisopliae var. majus are identical to those of M. anisopliae var. anisopliae with an exception for the presence of an additional site in the PCR product. Results from this study provide an additional method for identification and differentiation of isolates of these two varieties of M. anisopliae for use in the field and laboratory experiments.

• 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.

• Microbiology and Biotechnology Letters
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• v.23 no.6
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• pp.665-670
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• 1995

A marine bacterium which produces extracelluar agarase was isolated from sea water. Isolated strain was identified as Pseudomonas sp. by the morphological and biochemical properties (1). HindIII restriction fragment of 3.2 kb from Pseudomonas genomic DNA was cloned into pUC19 to obtain recombinant plasmid pJA1 which enables E. coli JM83 to produce agarase. Most of agarase produced in E. coli was secreted into the culture medium. The enzyme (pJA1) showed the highest agarase activity during the stationary phase (20 hrs) of E. coli. The optimum temperature and pH were 40$\circ$C and 7.8, respectively. Restriction gene map anlaysis revealed that it has different restriction pattern with three kind of agarase gene reported.

• Korean Journal of Agricultural Science
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• v.44 no.3
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• pp.318-324
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• 2017

With the development of next-generation sequencing (NGS), a cutting-edge technology, genotype-by-sequencing (GBS) became available at a low cost per sample. GBS makes it possible to customize the process of library preparation to obtain high-quality single nucleotide polymorphisms (SNPs) in the most efficient way. However, a GBS library is hard to construct due to fine-tuning of concentration of each reagent and set-up. The major reason for this is the presence of undigested genomic DNA (gDNA) owing to the efficiency of different restriction enzymes for different species with unknown reasons. Therefore, this proof-concept study is to demonstrate the unpredictable patterns of enzyme digestion from various plants in order to make the reader aware of the caution needed when choosing restriction enzymes for their GBS library preparations. Indeed, no pattern was found for the digestibility of gDNA samples and restriction enzymes in the current study. We suggest that more data should be accumulated on this matter to help researchers who want to apply GBS technologies in a variety of genetic approaches.

• The Plant Pathology Journal
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• v.27 no.2
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• pp.148-155
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• 2011

Our previous sequence and phylogenetic analyses of the Korean Rice stripe virus (RSV) suggested possible genetic reassortment of RNA segments, but whether this RNA variation contributed to the recent RSV outbreaks in Korea is yet unclear. To further clarify these RSV-RNA segment variations, we developed a reverse transcription-polymerase reaction/restriction enzyme (RT-PCR/RE) analysis-based method. We identified five REs, including DraI, EcoR1, NdeI/AseI, and SpeI, that could differentiate RSV RNA 1-4 subtypes, respectively. Our RT-PCR/RE results provided a clear pattern of RNA reassortment, i.e., different groups of isolates having their RNA segments derived from two to three different RSV ancestors, such as from Eastern and Southwestern Chinese or Japanese M and T isolates. We also found that the migratory small brown planthopper from Eastern China caught by aerial net traps that possesses RSV-RNA3 genotypes corresponds mainly to Eastern China, with a few for Southwestern China based on RT-PCR/RE, sequence and phylogenetic analyses, indicating that RSV populations in Eastern China may also have strong RNA variation. The development of an RE analysisbased method proved a useful epidemiological tool for rapid genotyping and identification of mixed infections by RSV strain and by different subtype.

• Journal of Korean Society of Forest Science
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• v.83 no.1
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• pp.20-24
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• 1994

In woody species with a long life span, the studies on inheritance of any trait may be very time consuming and laborious. Chloroplast DNA(cpDNA) has been a valuable tool in such studies since it has several unique features such as limited genome size and cytoplasmic inheritance. In the present study, cpDNAs from five different species of Populus(P. alba, P. glandulosa, P. alba${\times}$P. glandulosa, P. davidiana, and P. nigra), and Nicotiana tabacum were compared with regard to restriction fragment length polymophism. The results showed that cpDNAs among the species were very conserved, although some polymorphisms were observed when the DNAs were digested with restriction enzyme EcoRI or KphI. The other enzymes (Bgl II, and PstI) tested produced identical restriction fragmentation pattern among the species. However, cpDNAs from all the five Populus species showed different restriction fragmentation pattern from that of tobacco with the four restriction enzymes tested. Southern hybridization with tobacco rbcL gene fragment as a probe also produced identical pattern among Populus species. The results indicate that cpDNAs in the genus are very well conserved during evolution.

• Journal of Environmental Science International
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• v.5 no.5
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• pp.605-610
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• 1996

The bacteriophage from the fresh oyster, Crassostrea Virginica which is specific to the marine bacterium was isolated and characterized. Among the foci different vibrio species and the five different serotypes of Vibrio parahaemolyticus host strains tested, only two strains of the parahaemolyticus possessing K17, K52 antigens were highly sensitive to the phage. The size of the isolated plaque was 0.4mm and the electron microscopic head size of the isolated phage was about 67 nm long and 83 nm wide. PFU/ml was 1.25$\times$ $10^{11}$. The phase was sensitive to chloroform but resistant to acetone or methanol. The assay of the isolated phase nucleic acid was deoxyribonucleic acid. The restriction enzyme pattern showed 14 fragment from Hind III and 4 fragments from Eco R I. Two different antigenic groups showed-similar restriction enzyme patterns.

• Journal of Microbiology
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• v.45 no.2
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• pp.175-178
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• 2007

An endonuclease from Xanthomonas oryzae pathovar oryzae KACC 10331, XorII, was recombinantly produced in Escherichia coli using a T7 system. XorII was purified using a combination of ion exchange and immobilized metal affinity chromatography (IMAC). An optimized washing protocol was carried out on an IMAC in order to obtain a high purity product. The final amount of purified XorII was approximately 2.5 mg/L of LB medium. The purified recombinant XorII was functional and showed the same cleavage pattern as PvuI. The enzyme activity tested the highest at $25^{\circ}C$ in 50 mM NaCl, 10 mM Tris-HCl, 10 mM $MgCl_{2}$, and 1 mM dithiothreitol at a pH of 7.9.

• Environmental Mutagens and Carcinogens
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• v.22 no.3
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• pp.199-204
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• 2002

It has been known that Ganoderma lucidum and Lentinus edodes have anticancer activity and immune enhancing activity. These two mushrooms were grown in liquid culture and harvested. From these mycelia, DNA was isolated and EtBr-CsCl density gradient ultracentrifugation was performed to purify it further. Then mitochondrial DNA was isolated by bisbenzimide-CsCl density ultracentrifugaton. Mitochondrial DNA of Ganoderma lucidum was digested by restriction enzymes, EcoR I, Hind Ⅲ, and Pst I, then electrophoresed. It showed 12, 22, 4 fragments. Mitochondrial DNA of Lentinus edodes was digested by EcoR I. Electric pattern showed 6 fragments. 4 fragments had appeared by Pst 1 digested mitochondrial DNA. Hind ill couldn't digest mitochondrial DNA of Lentinus edodes. Mitochondrial DNA of fusants was isolated to compare to those of parents. The results showed that fusant P₂S₄has new, recombined mitochondrial DNA. But P₂S₄had the same DNA that Ganoderma lucidum had.