• Journal of Mushroom
• /
• v.7 no.4
• /
• pp.150-155
• /
• 2009

Because the import of Inonotus obliquus have been rapidly increased in Korea, we developed the Inonotus species-specific marker by using various sequences including ITS sequences, PCR-RFLP and STS primers and used this marker to determine both genetic relatedness and strains discrimination of Inonotus spp. Total 17 different Inonotus spp. were examined by using ITS sequences and classified into 2 different groups. One strain, ASI74008 isolated from Kamchaka island of Siberia, showed the high sequence identity (98%) at the nucleotide level to the other I. obliquus DSM strain, indicating the ASI74008 belong to I. obliquus species. Comparison of banding patterns after restriction enzyme digestions with PCR amplicons of ITS region revealed some variations depending on the species and strains. However, PCR products amplified with STS primer showed species specific patterns.Therefore, use of both STS primers and PCR-RFLP could help for better strain identification.

• Korean Journal of Microbiology
• /
• v.45 no.1
• /
• pp.17-25
• /
• 2009

We studied soil components, methane production, the number of methanogens, and T-RFLP patterns dependent on agricultural methods with the change of seasons. There is no regular increase or decrease tendency of the most soil components followed by sampling period. And the water content in soil was higher in October than May. Also a lot of methanogens existed in soil, and acetotrophs were relatively of smaller number than hydogenotrophs and formate utilizing methanogens using MPN (most probable number) enumeration. In the experiment using the formate, it was used from the first week, and only a minute amount was detecte after four weeks. However in the acetate, it was increased until the third week, and after that was consumed. And there was higher methane production for all soil samples which administered with the hydrogen spike. The activity of methanogens was higher in the organic and low-agrichemical agricultural method samples, and the organic agricultural method had high methanogen activity among the other samples. A result of T-RFLP pattern of mcrA gene digested with Sau96I, methanogen community have a little relation with agricultural methods and seasons. This results also agreed to no critical difference the soil components dependent on agricultural methods, but some analytical data have a positive relationship with a agricultural methods. Therefor we could concluded that the comparison study of community for soil bacteria sufficiently could be useful for the microbiological indicator.

• Journal of Periodontal and Implant Science
• /
• v.29 no.2
• /
• pp.349-355
• /
• 1999

The present study has been performed to develop a PCR technology to identify human immunoglobulin(Ig) allotypes with restriction fragment length polymorphism(RFLP) using a probe. Genomic DNA were ampilified with PCR tecnology using primers from peripheral blood lymphocytes of 10 periodontal patiens, whose Ig allotypes have been pre-determined by serological tecnique using heagglutination technique. The result indicated that the RFLP patterns could successfully differentiate the Ig allotypes, which suggests that this technology can be developed as a tool useful for population genetics studies.

• Journal of Microbiology
• /
• v.34 no.4
• /
• pp.295-299
• /
• 1996

To infer phylogenetic relationships between species of Trichaptum (Polyporaceae), RFLP analyses of PCR-amplified DNAs were accomplished. Regions coding for ITSs of nuclear SSU rRNA genes and for mitochondrial SSU rRNA genes from thirteen strains of four Trichaptum species (T. abietinum, T. biforme, T. fusco-violaceum, and T. laricinum) were amplified and digested with eight restriction enzymes. All the fragmentation patterns were characterized and coded as 0/1 for the absence/presence of fragments. A phylogenetic tree based on the combined data sets was constructed using the Dollo parsimony method. While every two strains of T. abietinum, T. biforme, T. fusco-violaceum, and T. laricinum formed an independent group, the other strains of T. abietimum and T. fusco-violaceum made mixed groupings among compared strains. It is inferred that T. abietinum and T. fusco-violaceum have more variations, possibly geographic or physiological ones, than other species in the genus.

• Journal of Microbiology and Biotechnology
• /
• v.22 no.4
• /
• pp.563-566
• /
• 2012

To investigate the possibility of horizontal gene transfer between agricultural microorganisms and soil microorganisms in the environment, Bacillus subtilis KB producing iturin and the PGPR recombinant strain Pseudomonas fluorescens MX1 were used as model microorganisms. The soil samples of cucumber or tomato plants cultivated in pots and the greenhouse for a six month period were investigated by PCR, real-time PCR, Southern hybridization, and terminal restriction fragment length polymorphism (T-RFLP) fingerprinting. Our data from Southern blotting and T-RFLP patterns suggest that the model bacteria do not give significant impacts on the other bacteria in the pots and greenhouse during cultivation.

• Korean Journal of Plant Resources
• /
• v.10 no.3
• /
• pp.250-257
• /
• 1997

The relationship of nine Medicago species belonging to four subgenera were analyzed by using SDS-PAGE and restriction fragment length polymorphism (RELP) methodologies. Sixty-eight bands of alcohol and salt soluble proteins and 85-133 RFLP markers were used to estimate the genetic distance among the species. These species were clustered together at around 0.1 to 0.4 level of distance for both kind of markers, indicating that Medicago species have a large genetic similarity. A combined cluster diagram, at a dissimilarity level of 0.3, differentiated nine species in four groups: group 1, M. littoralis , M. truncatulam, M.scutellata and M. rigidula; group 2, M. sativa ; group 3, M. lupulina ; group 4, M. orbicularis, M. radiata and M. minima. All of them, but except for M. minima. corrensponded to the existing four subgenera of the genus Medicago classified by Lesins and Lesins(1979).The most similar species were M. littoralis and M. trucatula and the most dissimilar one was M. lupulina. In separate cluster diagrams based on RFLP and protein markers, some differences were observed. In the case of RFLP or DNA markers, M. sativa (alfalfa) was distantly clustered with other Medicago species. But in the case of protein markers, M. sativa was closely clustered with M. scutellata, M. littorulis and M. truncatula.

• Journal of Mushroom
• /
• v.4 no.2
• /
• pp.43-47
• /
• 2006

PMO-P4, being cultivated as "Sanghwang" in Korea, was proved to be P. baumii based on ITS (internal transcribed spacer) sequencing and RFLP (Restriction Fragment Length Polymorphism) patterns along with some Phellinus species including P. linteus. The similaraty of ITS sequencing between PMO-P4 and other Phellinus species was given the range of 48.6%~72.2%, showing the highest homology from P. linteus and the lowest from P. gilvus.

• Fisheries and Aquatic Sciences
• /
• v.11 no.3
• /
• pp.133-139
• /
• 2008

Molecular techniques, including restriction fragment length polymorphism(RFLP) and polymerase chain reaction-single strand conformational polymorph isms(PCR-SSCP), were developed to identify salted, dried threadfin(Eleutheronema tetradactylum) and white herring(Ilisha elongata) fish. Using PCR with universal primers, conserved 367-bp fragments of the cytochrome b gene were amplified from fresh fish samples and sequenced. The sequences were then searched for specific restriction sites. The digestion of the PCR products with the endonucleases AvaI, FokI, MboII, and MspI generated RFLP, which was used to identify the commercial products. Similarly, the amplified PCR-SSCP products were developed and the products tested. Overall, similar patterns were found in the majority of the fresh and processed products. Based on the results, both RFLP and PCR-SSCP were useful in determining and validating the authenticity of the fish species used to prepare the commercial salted, dried products. A similar approach can be applied to other species.

• Korean Journal Plant Pathology
• /
• v.14 no.5
• /
• pp.519-525
• /
• 1998

Intraspecific genetic diversity of Korean isolates of Phytophthora drechsleri was investigated based on PCR-RFLP of rDNA along with closely related species in the genus; P. cryptogea, P. melonis, P. erythroseptica, P. cinnamomi, P. cambivora and P. cactorum. Gene regions of nuclear small subunit and internal transcribed spacer (ITS) in rDNA were amplified with polymerase chain reaction and digested with 9 restriction enzymes. Phytophthora species was readily differentiated from each other based on the digestion patterns, however, P. cryptogea was not separable from some isolates of P. drechsleri. Twenty one isolates of P. drechsleri originated from 15 host plants were divided into three distinct groups designated as PdG1, PdG2 and PdG3, respectively. Four isolates in PdG1 were originated from green vegetables and tomato and nine isolates in PdG2 were mainly isolated from medicinal plants. The two groups showed 95.3% homology and four isolates of P. cyptogea came under the groups. However, Eight isolates in PdG3 collected from cucurbits were clearly differentiated from those of PdG1 and PdG2 by 66.5% homology, but completely matched with a Taiwan isolate of P. melonis. Results indicated that three distinct groups exist in Korean isolates of P. drechleri and each group has host preference. In addition, reclassification of the cucurbits isolates are reserved because of their distinct genetic characters from other intraspecific groups in P. drechsleri.

• Journal of Food Hygiene and Safety
• /
• v.15 no.2
• /
• pp.128-136
• /
• 2000

Twenty-four Vibrio strains were isolated from imported frozen seafoods and identified according to their physiological and biochemical properties. They included two V cholerae non-01 sp., two V. diazotrophicus sp., one V. hollisae sp., five V. natriegens sp., eight V. fluvialis sp., and four V. nereis sp.. Two of them were not identified as Vibrio species. When these strains were tested using API-2OE kit fur identification, however, only the results for two V. cholerae and five of the V. fluvialis strains matched the results obtained previously. Due to the importance of detecting V cholerae from foods, phylogenetic identification of the strains was attempted based on restriction fragment length polymorphism (RFLP) of the 16S rDNAs amplified by PCR. The results suggested that the two strains had identical RFLP patterns which were more closely related to that of V. proteolyticus than V. cholerae. The problems associated with identification of pathogens originated from seafoods demand development of accurate and rapid identification methods.