• 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.

• 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 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 Plant Pathology Journal
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• v.36 no.4
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• pp.364-377
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• 2020

Sugarcane is an important sugar crop contributes more than 80% of world sugar production. Mosaic, leaf fleck, and yellow leaf (YL) are the major viral diseases affecting sugarcane, amongst YL occurrence is widely reported in all the sugarcane growing countries. It is caused by Sugarcane yellow leaf virus (SCYLV) and detailed works were done on complete genome characterization, transmission, and management. However, in countries like Egypt, South Africa, Cuba, Mauritius and Hawaii, the disease was reported to the cause of sugarcane yellow leaf phytoplasma (SCYP) and/or SCYLV as single/combined infections. Hence, we have investigated in detail to identify the exact Candidatus phytoplasma taxon associated in Indian cultivars affected with YL. The sequencing results and the restriction fragment length polymorphism pattern of the PCR products using the universal phytoplasma primers confirmed presence of sugarcane grassy shoot (SCGS) phytoplasma (16SrXI group) in the YL-affected plants. Mixed infection of SCYLV and SCGS phytoplasma was estimated as 32.8% in YL affected plants. Evolutionary genetic relationship between SCYP and SCGS phytoplasma representatively taken from different countries showed that SCYP from South Africa and Cuba were diverged from others and had a highest similarity with SCGS phytoplasma. Although we wanted to identify SCYP from YL affected Indian sugarcane cultivars, the study clearly indicated a clear absence of SCYP in YL affected plants and we found SCYLV as the primary cause for the disease.

• Journal of Plant Biology
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• v.39 no.1
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• pp.49-55
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• 1996

We have studied the DNA of Allium sativum L. with respect to highly repetitive sequences. Fast reassociated DNA fragments expected to be highly repetitive sequences based on $C_{o}t$ curve were isolated and characterized. Their copy numbers were approximately $10^{5}~10^{7}$ per haploid genome. Nucleotide sequences analysis of six candidates reveals that their G/C content were low, 25-40% and typical patterns of repeating sequences exist. Repeat sequences were used as probes to access restriction fragment length polymorphism (RFLP) of genomic DNAs of four local clones, Tanyang, Mungyong, So san, and Uisong. The hybridization pattern were very similar among these four local clones.clones.

• The Journal of the Korean Society for Microbiology
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• v.22 no.1
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• pp.35-53
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• 1987

Multiply resistant Shigella strains isolated in Taegu area were subjected for the characterization of R plasmids. All strains isolated in 1984 and 1985 were susceptible to gentamicin, amikacin, and cephalothin, and most strains were susceptible to kanamycin (Km) and rifampin by agar dilution antimicrobial susceptibility test. The resistance frequency of S. flexneri against ampicillin (Ap) was higher than that of S. sonnei. The strains resistant to sulfisomidine (Su) and trimethoprim (Tp) were found at higher frequency in S. sonnei than in S. flexneri. The most prevalent resistance pattern of S. flexneri was chloramphenicol (Cm) tetracycline (Tc) streptomycin (Sm) Ap, followed by the pattern of CmTcSmSuApTp, CmTcSmSuApTp nalidixic acid, and CmTcSmSuAp in the decreasing order. The antibiogram of CmTcSmSuTp was found to be the most frequent pattern in S. sonnei. The ratio of conjugal transfer of S. flexneri was 47% and 75% of S. sonnei. The average number of plasmid harboring in Shigella was 4 and the size of plasmid ranged 1.3 to 134 megadalton (Mdal). Most S. flexneri carried plasmids of 2 to 3 Mdal and S. sonnei carried those of 3 to 4 Mdal size. The sizes of conjugative plasmids ranged 40-90 Mdal. The incompatibility group (Inc) F II plasmids (54-59 Mdal) were most frequent and rare Inc B plasmids (60 Mdal) of isolates in 1979 and 1980 and Inc FI (87 Mdal) of 1983 isolates were able to be classified by the colony test with standard reference plasmids. The R plasmids of known Inc group were tested for the restriction endonuclease analysis. The pattern of plasmids digested by EcoRl were apparently different by the Inc group but there was no significant difference between species or by the resistance patterns. Nonconjugative plasmids and their phenotypes were identified by transformation test. The transformants were resistant to less than two drugs. Colicin producing transformants carried the Col plasmid of 3.7 or 3.9 Mdal size. $Ap^r$ plasmids derived from S. sonnei were found to be mobilized by transfer factor RT641 to E. coli #CS100. $Ap^r$ plasm ids of same size shared by S. flexneri, S. sonnei, and E. coli were digested with Pstl. All of them showed two restriction fragments of 2.8 kilobase(kb) and 0.7kb. Other plasmids ($Sm^r\;Su^r$) derived from S. flexneri, S. boydii, and S. sonnei were digested with Pstl and they showed same restriction fragment patterns of 3.1kb and 2.9kb. The plasmid profiles of three strains of S. sonnei producing colicin and showing same resistance pattern of CmTcSmSuApTpKm appeared to be similar. Restriction patterns by EcoRl and the behavior of plasmids in conjugation or transformation process were also similar between those plasmids. The restriction patterns were significantly different between the plasmids of Inc FI group and those of unclassified Inc group.

• Parasites, Hosts and Diseases
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• v.34 no.1
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• pp.15-20
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• 1996

Cysts of Entamoebn histoIMtica are still found from humans in Korea, but notall of the cysts are known as pathogenic. The non-pathogenic strain is regarded as a differenL species, E. nispnr. In this study, Korean isolates of conventional E. histolvticn were subjected for the differentiation by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analysis. Human stools were screened by routine microscopic examination, and cyst or trophozoite positive stools were inoculated into Robinson media. The cultivated trophozoites were prepared for DNA extraction, and the DNAs were used for PCR with common primers of Pl gene. The PCR products were divested with 3 restriction enzymes and RFLP was observed. Also anti-sense primers containing the cleavage site of each restriction eWe were designed for differentiation only by PCR. The PCR products of Korean isolates 59,512, YS-6, and YS-27 were spliced by Taq I and Xmnl but not byAccl, and the isolates S1, S3, S11, S15, S16, S17, S20, YS- l7, and YS-44 were spliced by Acc I but not by Taq I and Xmn I. These RFLP pattern correlated well with PCR products by the species specific primers. The findings confirm that the Korean isolates 59,512, YS-6, and YS-27 are E. histolwtico and others are E. dispar. In Korea, most of the asymptomatic cyst carriers are infected by E. dispar, not by E. histolytica. Key words: Entcmoebc histolytica, Entcmoebn dispar Korean isolates, polymerase chain reaction (PCR), restriction fragment length polymorphism (RFLP)

• Journal of Microbiology
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• v.34 no.1
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• pp.1-6
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• 1996

The nucleic acid of Synechococcus sp. cyanophage was identified as double-stranded DNA by the result of digestion with enzymes such as exonucleases, DNase, and S1 nuclease, and by acridine orange staining. The cyanophage DNA was cleaved with several restriciton ehdonucleases such as ApaI, BamHI, Bg/II, HaeIII, Eco RI, HindIII, PstI, AND aPAI gave the clearest sets of bands on agarose gels and the fragment numbers for each were 12, 20, 29, 20, and 7, respectively. The sums of the size from Bam HI and PstI digestions were estimated approximately 227$\pm$4 kb, which are in agreement with the result of the pulsed field gel electrphoresis. This virus is thought to have the largest genosome among those of known cyanophages, which corresponds to the largest haed ot 90 nm when compared with the head sizes of cyanophages discovered since 1963.

• Journal of Microbiology
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• v.35 no.3
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• pp.172-176
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• 1997

The Mycobacterium tuberculosis clinical isolates in Korea, showing different drug resistances, were analyzed by comparing large restriction fragment (LRF) patterns produced y digestion of genomic DNA with infrequent-cutting endonucleases of SpeI, AsnI and pulsed-field gel electrophoresis (PFGE). SpeI and AsnI allowed with AsnI and SpeI, strains yielded an absolutely identical pattern for Korean type's mycobacteria even though they showed different drug resisstance. However, when three M. tuberculosis strains, showing drug resistance, were digested with XbaI, patterns were different from those of the other M. tuberculosis strians which are susceptible to drugs. This stuyd reveals that the comparison of chromosomal restriction patterns is very useful as an additional aid for the differentiation and identification of M. tuberculosis strains showing drug resistances.

• The Plant Pathology Journal
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• v.22 no.1
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• pp.36-45
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• 2006

Twenty isolates of Didymella bryoniae were isolated from infected cucurbit plants in various growing areas of southern Korea in 2001 and 2002. Random Amplified Polymorphic DNA (RAPD) group [RG] I of D. bryoniae was more virulent than RG IV to watermelon. Virulence of the RG I isolate was strong to moderate to cucumber, whereas that of the RG IV varied from strong, moderate to weak. Two hundred seventy-three amplified fragments were produced with 40 primers, and were analyzed by a cluster analysis using UPGMA method with an arithmetic average program of NTSYSPC. At the distance level of 0.7, two major genomic DNA RAPD groups were differentiated among 20 isolates. The RG I included 7 isolates from watermelon and one isolate from melon, whereas the RG IV included 12 isolates from squash, cucumber, watermelon and melon. Amplification of internal transcribed spacer (ITS) region and small subunit rRNA region from the 20 isolates yielded respectively a single fragment. Restriction pattern with 12 restriction enzymes was identical for all isolates tested, suggesting that variation in the ITS and small subunit within the D. bryoniae were low. Amplification of the genomic DNAs of the tested isolates with the sequence characterized amplified regions (SCAR) primer RG IF-RG IR specific for RG I group resulted in a single band of 650bp fragment for 8 isolates out of the 20 isolates. Therefore, these 8 isolates could be assigned into RG I. The same experiments done with RG IIF-RG IIR resulted in no amplified PCR product for the 20 isolates tested. An about 1.4 kb-fragment amplified from the RG IV isolates was specifically hybridized with PCR fragments amplified from genomic DNAs of the RG IV isolates only, suggesting that this PCR product could be used for discriminating the RG IV isolates from the RG I isolates as well other fungal species.