• Mycobiology
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• v.42 no.1
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• pp.46-51
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• 2014

A degenerated strain of Pleurotus eryngii KNR2312 was isolated from a commercial farm. Random amplified polymorphic DNA analysis performed on the genomic DNA of the normal and degenerated strains of this species revealed differences in the DNA banding pattern. A unique DNA fragment (1.7 kbp), which appeared only in the degenerated strain, was isolated and sequenced. Comparing this sequence with the KNR2312 genomic sequence showed that the sequence of the degenerated strain comprised three DNA regions that originated from nine distinct scaffolds of the genomic sequence, suggesting that chromosome-level changes had occurred in the degenerated strain. Using the unique sequence, three sets of PCR primers were designed that targeted the full length, the 5' half, and the 3' half of the DNA. The primer sets P2-1 and P2-2 yielded 1.76 and 0.97 kbp PCR products, respectively, only in the case of the degenerated strain, whereas P2-3 generated a 0.8 kbp product in both the normal and the degenerated strains because its target region was intact in the normal strain as well. In the case of the P2-1 and P2-2 sets, the priming regions of the forward and reverse primers were located at distinct genomic scaffolds in the normal strain. These two primer sets specifically detected the degenerate strain of KNR2312 isolated from various mushrooms including 10 different strains of P. eryngii, four strains of P. ostreatus, and 11 other wild mushrooms.

• Clinical and Experimental Reproductive Medicine
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• v.24 no.1
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• pp.51-56
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• 1997

Recently, through the development of the primer extension preamplification(PEP) method which amplifies the whole genome, simultaneous multiple DNA analysis has become possible. Whole genome from each single cell can be amplified using 15 base oligonucleotide random primer. The greatest advantage of PEP-PCR is the ability to investigate several loci simultaneously and confirm results by analysing multiple aliquots for each locus. This technique led to the development of preimplantation genetic disease diagnosis using blastomere from early embryo, sperm, polar body and oocyte. In this study, we applied PEP-PCR in 20 cases of single amniocyte and 20 cases of single chorionic villus cell for the clinical application of the prenatal and preimplantational genetic diagnosis. We analysed 7 gene loci simultaneously which are 46, 47 exons related to dystrophin gene, two VNTR (variable number tandem repeat) markers using 5'dysIII, 3'CA related to dystrophin gene and DYZ1, DYZ3, DYS14 regions on chromosome Y. In all the tests, 97.5% of PEP-PCR amplifications with single cells were successful. We obtained 38/40 (95%) accuracy in gender determination through chromosome analysis comparison. Therefore, these results have significant implications for a sperm or oocyte analysis and prenatal or preimplantational genetic diagnosis.

• Applied Biological Chemistry
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• v.42 no.1
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• pp.34-38
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• 1999

We observed the structure of phenylalanine ammonia-lyase gene (PAL) which is one of the best studied plant defense-related genes responding to pathogen infection by producing suberin, lignin, and phytoalexins. In tomato, at least 5 different genetic loci have been identified by genomic southern blot hybridization and nucleotide sequence analyses of partially cloned gene fragments (Lee et al. 1992). However, our results suggest that two other isoforms designated as PAL X1 and PAL X2 are located on the chromosome in tomato plant. Furthermore, the preliminary results obtained from southern blot hybridization analyses of subcloned fragment digested with several restriction endonuclease indicated that PAL X1 and PAL X2 clones contain at least two copies of PAL gene and partial nucleotide sequence analyses of each subcloned fragment with the same primer taken from known nucleotide sequence of PAL5 gene indicated that they are located side by side on the same chromosome.

• The Plant Pathology Journal
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• v.23 no.1
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• pp.13-21
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• 2007

Sweet potato feathery mottle virus(SPFMV) is one of the most prevalent viruses infecting sweet potatoes and occurs widely in sweet potato cultivating areas in Korea. To assess their genetic variation, a total of 28 samples infected with SPFMV were subjected to restriction fragment length polymorphism(RFLP) analysis using DNAs amplified by RT-PCR with specific primer sets corresponding to the coat protein(CP) region of the virus. The similarity matrix by UPGMA procedure indicated that 28 samples infected with SPFMV were classified into three groups based on the number and size of DNA fragments by digestion of CP-encoding regions with 7 enzymes including SalI, AluI, EcoRI, HindIII, FokI, Sau3AI, and DraI bands. Four primer combinations out of 5 designed sets were able to differentiate SPFMV and sweet potato virus G infection, suggesting that these specific primers could be used to differentiate inter-groups of SPFMV. Sequence analysis of the CP genes of 17 SPFMV samples were 97-99% and 91-93% identical at the intra-group and inter-groups of SPFMV, respectively. The N-terminal region of the CP is highly variable and examination of the multiple alignments of amino acid sequences revealed two residues(residues 31 and 32) that were consistently different between SPFMV-O and SPFMV-RC.

• The Korean Journal of Mycology
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• v.38 no.2
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• pp.105-111
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• 2010

Lentinus lepideus, known as train wrecker fungus, has been used for nutritional and medicinal purposes. Recently, commercial cultivation technique and a new cultivar of the mushroom were developed. To investigate the genetic diversity and phylogenetic relationship for identifying the mushroom strains and cultivar, one commercial and 13 strains of Lentinus lepideus from different geographical regions of Korea were analyzed by ITS regions of rDNA and RAPD of genomic DNA. Three strains of Lentinus edodes were also used for the analysis. The size of the ITS1 and ITS2 regions of rDNA from the different strains varied from 173 to 179 bp and 203 to 205 bp, respectively. The sequence of ITS1 was more variable than that of ITS2, while the 5.8S sequences were identical with 156 base pairs. A phylogenetic tree based on the ITS region sequences indicated that selected strains could be classified into four clusters, while 3 strains of L. edodes was divided into a new cluster. Ten primers out of 20 arbitrary primers used in the RAPD-PCR efficiently amplified the genomic DNA. The numbers of amplified DNA bands varied with the primers and strains, with polymorphic DNA fragments in the range from 0.2 to 2.6 kb. The results showed that phylogenetic relationship among Korean strains of Lentnus lepideus is high, but genetic diversity is low.

• Research in Plant Disease
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• v.25 no.1
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• pp.16-21
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• 2019

Botrytis cinerea is the pathogen for a gray mold generating problems during the cultivation and transportation of roses. But there is little information about the difference of the symptom severity caused by gray mold on rose varieties and pathogen strains. 16 strains were collected from the rose cultivation area to confirm the degree of disease occurrence against strains and each variety. Collected 16 strains were identified based on the sequences analysis of ITS region of ribosomal DNA by using specific primers. The sequence analysis was performed by comparing the sequences to find a difference. To confirm the difference in disease occurrence for each strains, the difference was classified from 0 to 5 stages using charmant variety as a control. The data was confirmed through Kruskal-Wallis ANOVA. The result showed the significant difference in the pathogenicity caused by strains. WNG6_5 showed the lowest pathogenicity with 0.24 and WNG6_3 showed the highest with 3.20. The difference between two strains were almost 3.0. In addition, nine varieties of roses were more investigated with three strains such as the strains of WNG6_5, Hwa_1, and WNG6_3. The result showed that the Love Letter variety showed resistance and the Ice Bear variety was sensitive to three strains. Taken together, this study showed the significant difference by the interactions of rose varieties and gray mold strains.

• Korean Journal of Microbiology
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• v.27 no.1
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• pp.1-9
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• 1989

The locations of 5' ends as well as the splicing pattern of viral poly A(-) 19S RNA from monkey cells infected with SV40 were determined by a modification of primer extension method. The 5' end of this RNA mapped at the major cap site at nucleotide residue 325, used most frequently by SV40 late RNAs. The intron from nt.373 to nt.558 was removed as the ordinary cytoplasmic poly A(+) 19S RNA. The 3'end of this RNA was very heterogeneous and distributed over 1 kb upstream of polyadenylation site, as determined by S1 nuclease mapping. The reason for this normally initiated and spliced RNA to accumulate in the nucleus was investigated. In order to test whether the presence of unused 3' splice region on this RNA caused such subcellular distribution, cells were transfected with SV40 mutant KNA containing deletion around 3' splice site. The RNA deleted of 3' splice region accumulated mainly in the cytoplasm. This accumulation did not result from the increased stability of the RNA due to the deletion, since the wild type and mutant RNAs exhibited similar half lives after chase with actinomycin D. Therefore it is likely that the 19S spliced RNA is hindered from being transported into the cytoplasm due to some pre-splicing complexes formed at the unused 3' splice site.

• Journal of Life Science
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• v.24 no.2
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• pp.181-185
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• 2014

We identified SNPs (single nucleotide polymorphisms) for endothelial nitric oxide synthase (eNOS) genes in the Korean genome. eNOS is present in the vascular endothelium, platelets, and several other cell types that continuously produce modest amounts of NO. Endothelium-derived NO plays a key role in the regulation of vascular tone, and the impaired effects of NO on the cardiovascular system appear to be responsible for coronary atherosclerosis and thrombosis. In recent studies, a missense variant within exon 7 of the eNOS gene in patients with coronary spastic angina-GAG to GAT substitution, which results in the replacement of glutamic acid by aspartic acid (Glu298Asp [G894T])-has been identified and is known to be significantly associated with coronary spasm. We prepared PCR primers based on sequences in Genbank. Primers were prepared for normal and SNPs separately, as reported for other Asian countries, such as G894T. Their sequences were different only at the 3' ends so that primer extension could only by possible when base pairs between templates and primers matched. We also employed ARMS (Amplification Refractory Mutation System) technology to improve the specificity of the PCR reaction. In conclusion, we were able to demonstrate the eNOS G894A polymorphism in Korean gemone. This study should facilitate research on the cause of myocardial infarction and development on further therapy at the genetic level.

• The Korean Journal of Mycology
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• v.27 no.2 s.89
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• pp.132-137
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• 1999

This study was carried out to develop the molecular marker for the detection of Pseudomonas tolaasii, a causative agent of bacterial brown blotch disease of oyster mushroom (Pleurotus ostreatus). When several primers designed from repetitive sequences and pectin lyase genes of bacteria were used to produce DNA polymorphism from different Pseudomonas spp. isolated from edible mushrooms, PEU1 primer derived from pectin lyase gene produced polymorphic bands differentiating P. tolaasii strains from other Pseudomonas species. Two bands, 1.0kb and 0.4kb, found commonly in 6 isolates of P. tolaasii were cloned into pGEM-T vector which were designated as pPTOP1 and pPTOP2, respectively, to use as probe. The 0.4 kb insert of pPTOP2 hybridized to only 6 isolates of P. tolaasii, but did not to the other Pseudomonas species. As few as $1.5{\times}10^3$ colony forming unit (cfu) of P. tolaasii could be detected by dot blot hybridization with the cloned 0.4kb DNA in pPTOP2.

• Korean Journal of Medicinal Crop Science
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• v.11 no.5
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• pp.377-384
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• 2003

Genetic differences among nine land races of Korean ginseng (Panax ginseng C. A. Meyer) were examined using RAPD markers. Land races of Korean ginseng were collected from nine regions in Korea: Cheongwon, Guesan, Geumsan, Namwon, Pochun, Yangju, Yeoncheon, Yeongju. Out of 48 RAPD primers tested, 5 primers (OPA 7, OPA 13, URP 2, URP 3 and UBC 3) produced remarkable bands which showing polymorphisms among evaluated collections. Lower levels of genetic diversity were in detected same land races than among other land races. Genetic differences within and among land races indicate heterogeneity. These results indicate that cultivated ginseng in Korea is heterogeneous. Genetic similarity matrices of RAPD profiles were generated via coefficients of variation and the data were processed by the cluster analysis (UPGMA). When 90 collections were evaluated using selected 5 primers, those were clustered to 5 and 3 subgroups. These differences in genetic variation between land races of Korean ginseng implied the potential source for further breeding of Korean ginseng.