• The Korean Journal of Malacology
• /
• v.31 no.4
• /
• pp.257-262
• /
• 2015

The seven selected primers BION-13, BION-29, BION-61, BION-64, BION-68, BION-72 and BION-80 generated the total number of loci, average number of loci per lane and specific loci in Meretrix lusoria (ML), Saxidomus purpuratus (SP) and Cyclina sinensis (CS) species. Here, the complexity of the banding patterns varied dramatically between the primers from the three venerid clam species. The higher fragment sizes (> 1,000 bp) are much more observed in the SP species. The primer BION-68 generated 21 unique loci to each species, which were ascertaining each species, approximately 150 bp, 300 bp and 450 bp, in the ML species. Remarkably, the primer BION-80 detected 7 shared loci by the three clam species, major and/or minor fragments of sizes 500 bp, which were matching in all samples. As regards average bandsharing value (BS) results, individuals from CS clam species (0.754) exhibited higher bandsharing values than did individuals from SP clam species (0.607) (P < 0.05). In this study, the dendrogram obtained by the seven oligonucleotides primers indicates three genetic clusters: cluster 1 (LUSORIA01-LUSORIA07), cluster 2 (PURPURATUS08-PURPURATUS14), cluster 3 (SINENSIS15-SINENSIS21). Among the twenty one venerid clams, the shortest genetic distance that displayed significant molecular differences was between individuals 18 and 20 from the CS species (genetic distance = 0.071), while the longest genetic distance among the twenty-one individuals that displayed significant molecular differences was between individuals LUSORIA no. 02 and PURPURATUS no. 09 (genetic distance = 0.778). Relatively, individuals of SP venerid species were appropriately closely related to that of CS species, as shown in the hierarchical dendrogram of genetic distances. Eventually, PCR fragments exposed in the present study may be worthwhile as a DNA marker the three venerid clam species to discriminate.

• Korean Journal of Animal Reproduction
• /
• v.23 no.4
• /
• pp.303-311
• /
• 1999

Nuclear DNA was isolated from the sperm cells representing genetic characteristics and genomic polymorphisms of rainbow trout by polymerase chain reaction(PCR) amplification of DNA using arbitrary primers. Genomic DNA fingerprints were generated from rainbow trout sperm DNA by polymerase chain reaction amplification using 20 arbitrary decamers as primers. Out of these primers, 4 generated 17 highly reproducible RAPD markers, producing almost six polymorphic bands per primers. Four of 6 primers tested generated amplified fragments which were polymorphic between different individuals. Polymorphic DNA fragments were reproducibly amplified from independent DNA preparations made from individuals. Rainbow trout was distinctly observed 3 specific DNA markers (2. 3, 2.0 and 1.3kb) in bandsharing. Individual fragments generated using the same arbitrary primer, demonstrated that a single primer detected at least three independent genomic polymorphisms in rainbow trout sperm DNA. The RAPD polymorphism generated by this primer may be used as a genetic marker for individual identification The RAPD-PCR technique has been shown to reveal informative polymorphism in many species of fish. The present results demonstrate that RAPD markers are abundant, reproducible and provide a basis for future gene mapping and MAS in these important aquaculture species using RAPD polymorphic markers. It is concluded that RAPD polymorphisms are useful as genetic markers for fish breed differentiation.

• Journal of Microbiology and Biotechnology
• /
• v.10 no.5
• /
• pp.651-655
• /
• 2000

Polymerase chain reaction (PCR) was used to specifically detect Phytophthora infestans by analyzing the sequences of the ribosomal internal transcribed spacer regions (ITS) in the rDNA of the Phytophthora species. Based on the sequence data, PISP-1 together with the ITS3 primer were used to detect p. infestans. A single ca. 450 bp segment was observed in P. infestans, but not in the other fungal or bacterial isolates. Two factors, the annealing temperature and template DNA quantity, were investigated to determine the optimal conditions. Using these species-specific primers, a unique band was obtained within annealing temperatures of $55^{\circ}C$-$61^{\circ}C$ and template DNA levels of 10 pg-100 ng.

• The Plant Pathology Journal
• /
• v.18 no.6
• /
• pp.338-341
• /
• 2002

The potato scab is caused by several species of Streptomyces. Among these species, only pathogenic strains were found to produce thaxtomin A characterized by necrotic bioassay and HPLC. In this study, identification of the pathogenic strains of Streptomyces was performed through the polymerase chain reaction (PCR) by using specific pathogenicity primer sets derived from the nec1 gene sequences of Streptomyces scabies. The expected PCR products were obtained approximately 580 bp and confirmed by sequencing. This PCR technique can be used effectively to identify the pathogenic Streptomyces species, that cause scab on potato tubers.

• The Plant Pathology Journal
• /
• v.16 no.5
• /
• pp.252-256
• /
• 2000

The 16S-23S rRNA intergenic spacer regions (ISRs) were sequenced and analyzed to design specific primer for identification of Pectobacterium chrysanthemi. Two types ISRs, large and small ISRs, were identified from three strains (ATCC 11663, KACC 10163 and KACC 10165) of P. chrysanthemi and Pectobacterium carotovorum subsp. carotovorum ATCC 15713.Large ISRs contained transfer RNA-Ile(tRNA$^{Ile}$)and tRNA$^{Ala}$, and small ISRs contained tRNA$^{Glu}$. Size of the small ISRs of P. chrysanthemi ranged on 354-356 bp, while it was 451 bp in small ISR of P. carotovorum subsp. carotovorum ATCC 15713. From hypervariable region of small ISRs, species-specific primer for P. chrysanthemi with 20 bp length (CHPG) was designed from hypervariable region of small ISRs, which was used as forward promer to detect P. chrysanthemi strains with R23-1R produced PCR product of about 260bp size (CHSF) only from P. chrysanthemi strains, not from other Pectobacterium spp. and Erwinia spp. Direct PCR from bacterial cell without extracting DNA successfully amplified a specific fragment, CHSF, from P. chrysanthemi ATCC 11663. The limit of PCR detection was 1${\pm}10^2$ cfu/ml.

• Journal of the Korean Society of Food Culture
• /
• v.34 no.2
• /
• pp.208-216
• /
• 2019

Leuconostoc spp. are generally utilized as kimchi starters, because these strains are expected to have beneficial effects on kimchi fermentation, including improvement of sensory characteristics. Here, we developed a detection method for verifying the presence of the kimchi starter Leuconostoc mesenteroides WiKim33, which is used for control of kimchi fermentation. A primer set for multiplex polymerase chain reaction was designed based on the nucleotide sequence of the plasmids in strain WiKim33, and their specificity was validated against 45 different strains of Leuconostoc spp. and 30 other strains. Furthermore, the starter strain consistently tested positive, regardless of the presence of other bacterial species in starter kimchi during the fermentation period. Our findings showed that application of a strain-specific primer set for strain WiKim33 presented a rapid, sensitive, and specific method for detection of this kimchi starter strain during natural kimchi fermentation.

• Korean Journal of Veterinary Research
• /
• v.34 no.1
• /
• pp.115-125
• /
• 1994

In this study, we try to establish the rapid and specific detection system for Salmonella species. The PhoE gene of Salmonella species was amplified with two specific primers, ST5 and ST8c, using PCR. The probe prepared from the amplified PhoE gene was sequenced and applied for Southern blot analysis. After PCR with ST5 and ST8c primers for PhoE gene, DNA bands of expected size(365bp) from 7 different Salmonella species were detected, but not from 12 enterobacteriaceae and 3 gram positive bacteria. PCR was highly sensitive to detect up to 10fg of purified DNA template and to identify Salmonella species with only 320 heat-lysed bacterial cells. The inhibition of PCR amplification from stool specimen was occurred with 50-fold dilution but disappeared over 100 fold dilution of samples. It was confirmed that the PhoE genes were amplified and cloned with over 97% nacleotide sequence homology of PCR products compared with that of S. typhfmurium LT2. The DNA probe derived from S. typhimurium TA 3,000 showed highly specific and sensitive reaction with PCR products of all tested Salmonella species. These results indicate that PCR was rapid and sensitive detection method for Salmonella species and DNA probe prepared from S. typhimurium TA 3,000 was specific to identify PCR products of different Salmonella species.

• Journal of Applied Biological Chemistry
• /
• v.52 no.1
• /
• pp.1-7
• /
• 2009

For the development of qualitative PCR detection method of genetically modified (CM) rice, rice species-specific gene, OsCc-1 (rice cytochrome c gene), was selected as suitable far use as an endogenous gene in rice. The primer pair OsCytC-5'/3'with 111 bp amplicon was used for PCR amplification of the rice endogenous gene, OsCc-1 and no amplified product was observed from 8 different crops as templates. Qualitative PCR method was carried out with stack traits of L528$\times$CryIAc1 GM rice developed in Korea. For the qualitative PCRs, some primer pairs were designed with a construct-specific and event-specific type based on T-DNA and junction sequences of T-DNA in GM rice. Actck-5'/3' amplifying between actin promoter and OsCK1 gene introduced in LS28 gave rise to an amplicon 306 bp; also, CrLB-5'/3' from CryIAcl and CKRB-5'/3'amplifying the junction region of T-DNA and genome sequence from LS28 as event-specific primers gave rise to an amplicon 142 bp and 91 bp, respectively. These primer pairs for the detection of event-specific targets not produced PCR amplicons on non-CM rice and various crops in contrast to event lines. Therefore, in this study we verified that event-specific primers were effective to specifically detect stack trait lines and demonstrated that this method presented could be provided with the detection-method data for risk assessment analysis of GM rice to be commercialized.

• Mycobiology
• /
• v.32 no.3
• /
• pp.123-127
• /
• 2004

Sweet persimmons have been increasingly cultivated in the southern part of Korea. However, anthracnose disease caused by Colletotrichum species is one of the major hindrances in cultivation and productions. In this study, we used polymerase chain reaction(PCR) to detect Colletotrichum species with the AFLP(amplified fragment length polymorphism) method. In AFLP, we used E3(5'-GACTGCGTACCAATTCTA-3') and M1(5'-GATGAGTCCTGAGTAACAG-3') primer combination and, as a result, 262 bp segment was observed in Colletotrichum species only. Specific PCR primers were designed from the sequence data and used to detect the presence of the fungus in genomic DNA isolated from symptomless sweet persimmon plants. Based on sequence data for specific segments, Co.B1(5'-GAGAGAGTAGAATTGCGCTG-3') and Co.B2(5'-CTACCATTCTTCTA GGTGGG-3') were designed to detect Colletotrichum species. The 220 bp segment was observed in Colletotrichum species only, but not in other fungal and bacterial isolates.

• Journal of Practical Agriculture & Fisheries Research
• /
• v.24 no.3
• /
• pp.5-14
• /
• 2022

Trichoderma is known as pathogen caused serious green mold disease on commercial production. T. pleuroti and T. pleuroticola were common species in various mushroom media. Many strains of T. pleuroti, known as aggressive species causing major economic losses in Korea, showed benomyl resistance. Accurate identification and detection of Trichoderma species associated with oyster mushrooms is very important for disease control. We developed species-specific primers for T. pleuroticola, T. pleuroti, T. harzianum, and T. atroviride based on species-specific fragments isolated from amplified fragment length polymorphism analysis. PCR products corresponding to the predicted fragment of 500bp, 230bp, 180bp, and 410bp were amplified from T. pleuroticola, T. pleuroti, T. harzianum, and T. atroviride, respectively. Multiplex PCR assay using species-specific primers quickly and accurately identified and detected T. pleuroti from mushroom media in which various species co-exist. Our results can be useful for the effective control of mushroom disease.