• Interdisciplinary Bio Central
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• v.2 no.4
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• pp.12.1-12.7
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• 2010

The National Bio Resource Project-Rat (NBRP-Rat) comprises the largest bank of laboratory rat (Rattus norvegicus) strains in the world. Its main focus is to develop infrastructure that will facilitate the systematic collection, preservation, and provision of rat strains. To breed effectively more than 180 rat strains in living stock, we establish the genetic control system in which a systematic set of genetic diagnoses and genetic monitoring are included. Genetic monitoring is performed by using 20 polymorphic markers. Monitoring is carried out when a living animal stock is re-established by using cryopreserved embryos or sperm or when a rat strain is first introduced to the NBRP-Rat by a depositor. Additional monitoring is then carried out on each strain every two years. Genetic diagnosis is performed largely by employing the Amp-FTA method. Protocols which detail how to perform a genetic diagnosis of 11 transgenes and 24 mutations have been made. Among the mutations, nine can be detected by simple gel electrophoresis of the PCR products, 11 by restriction enzyme treatment of the PCR products, and four by direct PCR product sequencing. Using this genetic control system, the NBRP-Rat can guarantee the genetic quality of its rat strains.

• Korean Journal of Medicinal Crop Science
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• v.22 no.6
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• pp.429-434
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• 2014

This study describes the efficient method for the discrimination of 'Cheonryang' in Panax ginseng Meyer using a STS primer. A total of 208 STS primers were applied to polymerase chain reaction (PCR) amplification for discriminating Korean ginseng cultivars. Co-dominant polymorphic band patterns were generated with two primers, MFGp 0019, MFGp 0248, and successful identification of 'Cheonryang' was achieved from out of 11 Korean ginseng cultivars. Two different sizes of DNA band patterns were detected with MFGp 0019 primer. Ten Korean ginseng cultivars shared the same size of amplified DNAs (389 bp), but 'Cheonryang' showed a different size. Thus 'Cheonryang' can be efficiently distinguished from the other ten ginseng cultivars by using the MFGp 0019 primer. In the case of MFGp 0248, two different sizes of DNA band patterns were detected in the eleven ginseng cultivars. Same sized amplified DNA bands (307 bp) were shown in five cultivars (Chunpoong, Gopoong, Kumpoong, Cheongsun, Sunhyang) and 254 bp sized DNA bands were identified in the other 6 cultivars (Yunpoong, Sunpoong, Sunun, Sunone, Cheonryang, K-1). In conclusion, the two STS primers, MFGp 0019, and MFGp 0248, provide a rapid and reliable method for the specific identification of 'Cheonryang' cultivar from a large number of samples.

• Journal of Plant Biotechnology
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• v.29 no.4
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• pp.247-252
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• 2002

The combination of radiation technique with an in vitro culture system was initiated to develop salt tolerant rice. We established an in vitro culture system to select tolerant lines against salt stress. NaCl tolerant cell lines were selected from the callus irradiated with gamma ray on N$_{6}$ medium with 1.5% NaCl and 2 mg/L 2,4-D. Regenerants (M$_1$) were obtained from the tolerant callus which was cultured for 30 days auxin-free medium. The M$_2$seeds were harvested from M$_1$plants on an individual plant basis. Thirty seedlings from each 450 M$_2$lines were transplanted in a field and total 5,000 M$_3$lines were harvested with an average 90 percent of fertile grain. M$_3$lines were utilized for selection of salt tolerance. Salinity-tolerant lines (225) were selected among 5,000 M$_3$lines. Of the 225 lines tested, the morphological traits of two lines (120-10 and -11) were far superior to control (Donagjinbyeo) in agromomic traits such as plant height, root length and no. of roots. Control and tolerant lines were analyzed by RAPD markers. Three polymorphic bands were presented in only tolerant lines, demonstrating a genetic difference between control and the tolerant lines. Such tolerant lines could be used as genetic resources to improve salt tolerance.e.

• Journal of Mushroom
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• v.17 no.4
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• pp.171-178
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• 2019

Agaricus bisporus is a functional food and among the most widely cultivated mushrooms in the world. In this study, we analyzed the genetic diversity and population structure of 23 Korean and 42 foreign A. bisporus cultivars using SSR (Simple sequence repeat) markers. Genetic diversity of A. bisporus cultivars was as follows: number of alleles was approximately 13; observed and expected heterozygosity were approximately 0.59 and 0.74, respectively; and polymorphic information content value was about 0.71. A. bisporus cultivars were divided into three groups using distance-based analysis. Genetic diversity of Group 2, which consisted of cultivars from various countries, was high. In addition, model-based subpopulations were divided into two, and the genetic diversity of Pop2 (Population 2), which had many cultivars, was high. The results of this study could be used in a breeding program for A. bisporus, such as the development of new genetic resources and securing diversity.

• Horticultural Science & Technology
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• v.35 no.1
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• pp.98-107
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• 2017

A DNA profile database was constructed to investigate the genetic relatedness of 72 germplasm samples of Pyrus and related cultivars using microsatellite markers. Three P. pyrifolia, four P. commus, and one P. betulifolia cultivars with different morphological traits were screened using 387 pairs of microsatellite primers. A core set of 11 primer pairs was selected to obtain 133 polymorphic amplified fragments meeting three criteria: high polymorphism information contents (PIC), high repeatability, and distinct allele patterns. The number of alleles per locus ranged between 4 and 22. Average PIC was 0.743 (range: 0.557 - 0.879). Cluster analysis using the unweighted pair - group method with arithmetical average (UPGMA) separated the 72 pear cultivars and germplasm samples into four major groups: Chinese, European pears, and a cluster of 55 Asian pears that could be reclassify into two subcluster, I - $1^{st}$ and II - $2^{nd}$, according to pedigree information. Almost all of the cultivars were discriminated by 11 microsatellite marker genotypes. The microsatellite DNA profile database may be utilized as tool to verify distinctness, uniformity, and stability between candidate cultivar, and to verify in the distinctness of existing cultivars.

• Journal of Life Science
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• v.15 no.3 s.70
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• pp.474-478
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• 2005

This study was conducted to analyze genetic characteristics and to develop the specific marker for Cheju native horse (Coo) at the level of sequence characterized amplified regions (SCARs). We collected blood samples from Cheju native horse and Thoroughbred horse (Th) and obtained genomic DNA from the blood of 50 individuals randomly selected within the breeds. Seven hundred primers were chosen randomly and were used to examin the polymorphism and 40 kinds of primers showed polymorphic RAPD band patterns between two breeds. Thirty primers of them showed horse specific bands. With the primer MG 30, amplified band of 2.0 kb showed the specificity to Cheju native horse (Cnh). Additionally MG 53 detected the thoroughbred horse (Th) specific markers at size of 2.3 kb. As the next, 2.3 kb band from MG 53 was checked with the all individuals from all the breeds of this study, and it maintained the reproducible breed specificity to thoroughbred horse (Th). With this results, 2.3 kb band was cloned into plasmid vector and sequenced bidirectionally from both ends of the cloned fragment. With the obtained sequences 10 nucleotide extended primers including the original arbitray primer were designed as a SCARs primer. Finally, the primer with extended sequence showed the reproducible breed differentiation pattern and it was possible to identify Cheju native horse (Cnh) from other breeds. The SCARs marker 2.3 kb from MG 53 could be used to identify Cheju native horse (Cnh) for not only registration but also horse breeding programe.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.8
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• pp.1119-1126
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• 2018

Objective: Present investigation was aimed to study the Single Nucleotide Variants of the luteinizing hormone beta ($LH{\beta}$) gene and to analyze their association with the semen quality (fresh and post-thawed frozen semen) and luteinizing hormone (LH) concentrations in Murrah buffalo bulls. Methods: Polymerase chain reaction-single stranded conformational polymorphism (PCR-SSCP) and Sanger sequencing method is used to study genetic variability in $LH{\beta}$ gene. LH assay was carried out using enzyme-linked immunosorbent assay method. A fixed general linear model was used to analyze association of single nucleotide polymorphism (SNP) of $LH{\beta}$ gene with semen quality in 109 and LH concentrations in 80 Murrah bulls. Results: $LH{\beta}$ gene was found to be polymorphic. Total six SNPs were identified in $LH{\beta}$ gene g C356090A, g C356113T, g A356701G, g G355869A, g G356330C, and g G356606T. Single Stranded Conformational Polymorphism variants of pattern 2 of exon 1+pattern 2 of exon 2+pattern 1 of exon 3 had highly significant (p<0.01) effect on sperm concentration (million/mL), percent mass motility, acrosome integrity and membrane integrity in fresh and frozen semen whereas significant (p<0.05) effect was observed on percent live spermatozoa. SSCP variants of pattern 2 of exon 1+pattern 2 of exon 2+pattern 1 of exon 3 had highly significant (p<0.01) effect on luteinizing hormone concentrations too. Conclusion: The observed association between SSCP variants of $LH{\beta}$ gene with semen quality parameters and LH concentrations indicated the possibilities of using $LH{\beta}$ as a candidate gene for identification of markers for semen quality traits and LH concentrations in Murrah buffaloes.

• Korean Journal of Plant Resources
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• v.28 no.5
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• pp.648-655
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• 2015

Balloon flower (Platycodon grandiflorum A. DC.) is a perennial plant of mainly Campanulaceae family, which have been widely used as a food ingredient and herbal medicine in East Asia. Although demands on related products and yearly cultivation area for balloon flower are increasing, diverse fundamental technologies and molecular breeding studies are not very well supported in Platycodons. In this study, 30 random amplification of polymorphic DNA (RAPD) primers were test in an attempt to explore genetic diversities. In addition, sequences information of the actin gene, a well conserved gene encoding a globular protein that forms microfilaments, was retrieved and analyzed. Two actin homologs were recovered; 3.4 kb fragment is a Pg-actin and 1.4 kb fragment is a Pg-actin homolog with 28.6% similarity. We have confirmed that the Pg-actin gene is configured into 4 exons and 3 introns. A single nucleotide polymorphism (SNP), G↔A, was detected on the intron 3, which served as a target for the CAPS marker development. The marker Pg-Actin-Int3 was applied to 32 balloon flower accessions. Balloon flower DNA sequence information generated in this study is expected to contribute to the analysis and molecular breeding and genetic diversity analysis of balloon flowers.

• Journal of Korean Society of Forest Science
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• v.104 no.4
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• pp.549-557
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• 2015

Novel cpSSR primers were developed based on the sequence information of the Pinus koraiensis chloroplast genome. A total of 30 cpSSR loci were detected in the chloroplast genome, and a total of 30 primer sets flanking those loci were designed. All primer sets were successfully amplified for chloroplast DNA in P. koraiensis. The cross-species transferability of the 30 primer sets was considerably high in P. pumila (100%) and P. paviflora (97%) belonging to the same Subgenus (Strobus) of P. koraiensis. Meanwhile, the transferability was relatively low (73%) in P. densiflora and P. sylvestris belonging to Subgenus Pinus. A total of 13 cpSSR loci out of the 30 loci were polymorphic in the Mt. Jumbong population of P. koraiensis. The mean of haploid diversity(H) was 0.512. The number of haplotypes(N) and the haplotype diversity($H_e$) were 25 and 0.992, respectively. Of the 25 haplotypes, 22 were unique in the analyzed population. The unique haplotypes differentiated 22 individuals (79%) from the total of 28 individuals. In conclusion, the novel cpSSR primers developed in this study would be applicable to other Pinus species, especially the subgenus Strobus, and provide a high level of polymorphism for the study of genetic variation of P. koraiensis.

• Journal of Gastric Cancer
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• v.4 no.2
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• pp.109-120
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• 2004

Purpose: Individual gastric cancers demonstrate complicated genetic alterations. The PCR-based analysis of polymorphic microsatellite sequences on cancer-related chromosomes has been used to detect chromosomal loss and microsatellite instability. For the purpose of preoperative usage, we analyzed the correspondance rate of the microsatellite genotype between endoscopic biopsy and surgical specimens. Materials and Methods: Seventy-three pairs of biopsy and surgical specimens were examined for loss of heterozygosity and microsatellite instability by using 40 microsatellite markers on eight chromosomes. Microsatellite alterations in tumor DNAs were classified into a high-risk group (baselinelevel loss of heterozygosity: 1 chromosomal loss in diffuse type and high-level loss of heterozygosity: 4 or more chromosomal losses) and a low-risk group (microsatellite instability and low-level loss of heterozygosity: 2 or 3 chromosomal losses in diffuse type or $1\∼3$ chromosomal losses in intestinal type) based on the extent of chromosomal loss and microsatellite instability. Results: The chromosomal losses of the biopsy and the surgical specimens were found to be different in 21 of the 73 cases, 19 cases of which were categorized into a genotype group of similar extent. In 100 surgical specimens, the high-risk genotype group showed a high incidence of nodal involvement (19 of 23 cases: $\leq$5 cm; 23 of 24 cases: >5 cm) irrespective of tumor size while the incidence of nodal involvement for the low-risk genotype group depended on tumor size (5 of 26 cases: $\leq$5 cm; 18 of 27 cases: >5 cm). Extraserosal invasion was more frequent in large-sized tumor in both the high-risk genotype group ($\leq$5 cm: 12 of 23 cases; >5 cm: 23 of 24 cases) and the low-risk genotype group ($\leq$5 cm: 7 of 26 cases; >5 cm: 16 of 27 cases). The preoperative prediction of tumor invasion and nodal involvement based on tumor size and genotype corresponded closely to the pathologic tumor stage (ROC area >0.7). Conclusion: An endoscopic biopsy specimen of gastric cancer can be used to make a preoperative genetic diagnosis that accurately reflect the genotype of the corresponding surgical specimen.