• Food Engineering Progress
• /
• v.21 no.2
• /
• pp.174-179
• /
• 2017

For preliminary molecular typing, PCR-based fingerprinting using random amplified polymorphic DNA (RAPD) is the method of choice. In this study, 14 bacterial strains were isolated from different Korean food sources, identified using 16S rRNA gene sequencing, and characterized through RAPD-PCR. Two PCR primers (239 and KAY3) generated a total of 130 RAPD bands, 14 distinct PCR profiles, 10 polymorphic bands, one monomorphic band, and four unique bands. Dendrogram-based analysis with primer 239 showed that all 14 strains could be divided into seven clades out of which clade VII had the maximum of seven. In contrast, dendrogram analysis with the primer KAY3 divided the 14 L. citreum strains into four clades out of which clade IV consisted of a maximum of 10 strains out of 14. This research identified and characterized bacterial populations associated with different Korean foods. The proposed RAPD-PCR method, based on sequence amplification, could easily identify and discriminate the lactic acid bacteria species at the strain-specific level and could be used as a highly reliable genomic fingerprinting tool.

• Journal of Yeungnam Medical Science
• /
• v.22 no.2
• /
• pp.166-182
• /
• 2005

Background: Cyclin-dependent kinase (CDK) inhibitors are family of molecules that regulate the cell cycle. The CDKN2, a CDK4 inhibitor, also called p16, has been implicated in human tumorigenesis. The CDKN2 inhibits the cyclin/CDK complexes which regulate the transition from G1 to S phase of cell cycle. There is a previous report that homozygous deletion of CDKN2 region on chromosome 9p21 was detected frequently in astrocytoma, glioma and osteosarcoma, less frequently in lung cancer, leukemia and ovarian cancer, but not detected in colon cancer and neuroblastoma. However, little is known about the relationship between CDKN2 and laryngeal cancer. Therefore this study was initiated to investigate the role of CDKN2 in human laryngeal squamous cell carcinoma development.1) Materials and methods: We used 5 human laryngeal carcinoma cell lines whether they have deletions or losses of CDKN2 gene expression by DNA-PCR or RT-PCR, respectively. We examined 8 fresh frozen human laryngeal cancer tissues to detect the loss of heterozygosity (LOH) of CDKN2. PCR was performed by using microsatellite markers of short arm of human chromosome 9 (D9S126, D9S144, D9S156, D9S161, D9S162, D9S166, D9S171, D9S200 and D9SIFNA). For informative cases, allelic loss was scored if the signal of one allele was significantly decreased in tumor DNA when compared to the same allele in normal DNA. Results: The CDKN2 DNA deletion was observed in 3 cell lines. The CDKN2 mRNA expression was observed in only one cell line, which was very weak. LOH was detected in 7 cases (87.5%). Conclusion: These results suggest that CDKN2 plays a role in the carcinogenesis of human laryngeal squamous cell carcinoma.

• The Korean Journal of Mycology
• /
• v.44 no.2
• /
• pp.103-107
• /
• 2016

To establish a rapid and accurate detection of Phytophthora pinifolia, which is a quarantine pathogenic fungus in Korea, a species-specific primer was developed based on the ras-related protein (Ypt1) gene. Species-specific primer based on the DNA sequences of Ypt1 gene amplified 193 bp polymerase chain reaction (PCR) product for P. pinifolia. The primer pair yielded the predicted PCR product size exactly in testing with target pathogen DNAs, but not from the other 10 species of Phytophthora and 14 species of other phytopathogenic fungi. The primer pair also showed only the species-specific amplification curve on realtime PCR on target pathogen DNA. The detection sensitivity of real time PCR using species-specific primer pair was 10 to 100 times higher than conventional PCR, with 1 to $10pg/{\mu}L$.

• Proceedings of the Korean Society of Crop Science Conference
• /
• 2004.04a
• /
• pp.52-53
• /
• 2004

• The Korean Journal of Mycology
• /
• v.49 no.4
• /
• pp.455-467
• /
• 2021

Macrofungi are valuable resources as novel drug candidates, new biomaterials, and edible materials. Recently, genetic approaches pertaining to macrofungi have been continuously growing for their identification, molecular breeding, and genetic engineering. However, purification and amplification of fungal DNA is challenging because of the rigid cell wall and presence of PCR inhibitory metabolites. Here, we established a direct PCR method to provide a rapid and efficient method for PCR-grade macrofungal DNA preparation applicable to both conventional PCR and real-time PCR. We first optimized the procedure of lysis and PCR using the mycelia of Lentinula edodes, one of the most widely consumed macrofungal species. Lysates prepared by neutralizing with (NH₄)₂SO₄ after heating the mycelia in a mixture of TE buffer and KOH at 65℃ for 10 min showed successful amplification in both conventional and real-time PCR. Moreover, the addition of bovine serum albumin to the PCR mixture enhanced the amplification in conventional PCR. Using this method, we successfully amplified not only internal transcribed spacer fragments but also low-copy genes ranging in length from 500 to 3,000 bp. Next, we applied this method to 62 different species (54 genera) of macrofungi, including edible mushrooms, such as Pleurotus ostreatus, and medicinal mushrooms such as Cordyceps militaris. It was found that our method is widely applicable to both ascomycetes and basidiomycetes. We expect that our method will contribute to accelerating PCR-based approaches, such as molecular identification, DNA marker typing, gene cloning, and transformant screening, in macrofungal studies.

• Journal of Animal Science and Technology
• /
• v.51 no.3
• /
• pp.201-206
• /
• 2009

Seventeen porcine microsatellite (MS) markers recommended by the EID+DNA Tracing EU project, ISAG and Roslin institute were selected for the use in porcine individual and brand identification. The MSA, CERVUS, FSTAT, GENEPOP and API-CALC programs were applied for calculating heterozygosity indices. By considering the hetreozygosity value and PCR product size of each marker, we established a MS marker set composed of 13 MS markers (SW936, SW951, SW787, S00090, S0026, SW122, SW857, S0005, SW72, S0155, S0225, SW24 and SW632) and two sexing markers. The expected probability of identity among genotypes of random individuals (PI), probability of identity among genotypes from random half sibs ($PI_{half-sibs}$) and among genotypes of random individuals, probability of identity among genotypes from random sibs($PI_{sibs}$) were estimated as $2.47\times10^{-18}$, $6.39\times10^{-13}$ and $1.08\times10^{-8}$, respectively. The results indicate that the established marker set can provide a sufficient discriminating power in both individual and parentage identification for the commercial pigs produced in Korea.

• Proceedings of the Korean Society for Food Science of Animal Resources Conference
• /
• 2004.10a
• /
• pp.132-136
• /
• 2004

본 연구는 근육 및 지방세포 분화에 관여하는 leptin, MYF5 및 H-FABP의 3개 기능성 후보 유전자가 한우의 도체특성 및 육질에 미치는 영향을 분석하기 위하여 이들 유전자의 PCR-RFLP marker와 도체형질과의 관련성을 분석하였다. Leptin, MYF5 및 H-FABP 유전자에서 AA, AB 및 BB 3종류의 RFLP 유전자형이 각각 검출되었고 A와 B 대립유전자 빈도는 각각 0.57과 0.43, 0.61과 0.39 그리고 0.90과 0.10으로 추정되었다. 육질 등급에 따라 고급육과 저급육으로 분리 선발한 두 그룹간의 대립유전자 출현빈도를 비교한 결과 leptin과 MYF5 유전자에서 각각 통계적 유의차(P< .05)가 인정되었다. 또한 각 후보유전자의 RFLP marker 유전자형이 도체형질에 미치는 효과를 분석한 결과 leptin 유전자는 등지방 두께 그리고 MYF5유전자는 배장근 단면적에 각각 유의적인 영향(P< .05)을 미치는 것으로 분석되었다. 그러나 H-FABP 유전자는 도체형질들과 유의성이 인정되지 않았다. 따라서, leptin과 MYF5 유전자는 한우의 도체특성 및 육질 개선을 위한 DNA marker로 이용 가능할 것으로 사료된다.

• Proceedings of the Korean Society for Food Science of Animal Resources Conference
• /
• 2006.05a
• /
• pp.117-121
• /
• 2006

본 연구는 한우의 도체 품질을 결정하는 육질 등급 판정 항목이자 경제적으로 매우 중요한 근내지방도, 배최장근 단면적 및 등지방두께에 대한 개체별 유전능력 차이를 조기에 식별하는 선발 기술을 개발하기 위해 세포내 지방산 농도 및 다양한 세포 및 지질대사를 조절하는 지 방산결합 단백질(H-FABP) 유전자의 전체 염기서열을 분석하여 SNP를 검출하고, SNP marker가 한우의 도체 및 육질 형질에 미치는 영향에 대하여 분석하고자 수행하였다. 염기 서열 분석 결과 총 6개의 SNP를 검출하였고, 이들 중 4개의 주요 SNP를 선발하여 PCR-RFLP기법으로 genotyping하고, 각 SNP marker가 한우 도체 및 육질 형질에 미치는 영향을 통계분석 하였다. H-FABP의 C3523T SNP marker가 한우의 배최장근 단면적 및 등지방 두께에 유의적인 영향을 미쳤다(p<0.05). 따라서, 본 연구를 통해 개발된 한우 H-FABP 유전자의 특정 SNP marker는 배최장근 단면적은 넓고 등지방두께는 얇은 우수한 고급육을 생산하는 한우의 조기 식별 및 육질진단에 매우 유용한 SNP분자 표지로 활용할 수 있을 것으로 기대된다.

• The Korea Journal of Herbology
• /
• v.27 no.6
• /
• pp.15-21
• /
• 2012

Objective : Lonicera japonica THUNB. a traditional herbal medicine, has been commonly used anti-inflammatory disease. It has been very complicated with respect to its sources on the market. The significant selection of medicine depends on its origin. However, it is difficult to discrimination criteria for confirming L. japonica authenticity using the senses. This study was performed to determine the discriminant analysis of L. japonica and L. confusa. Methods : The identification of L. japonica and L. confusa were performed by the classification and identification committee of the national center for standardization of herbal medicines. And we examined its differences using HPLC and genetic marker analysis. Results : The analytical pattern of High Performance Liquid Chromatography was determined from the corresponding peak curves ((E)-aldosecologanin, chlorogenic acid, luteolin 7-O-glucoside, sweroside). For L. japonica, additional unknown peaks were detected at 13.8 min, 20.6 min, and 36.9 min. And, we developed genetic marker using the the tRNA-Leu gene, trnL-trnF intergenic spacer and tRNA-Phe region of chloroplast DNA. By the method, 164 bp PCR product amplified from L. confusa was distinguished into L. japonica and L. confusa efficiently. Conclusion : Base on these results, two techniques provide effective approaches to distinguish L. japonica from L. confusa.

• The Korean Journal of Mycology
• /
• v.46 no.2
• /
• pp.186-192
• /
• 2018

Rapid and accurate detection methods for Colletotrichum coccodes, an anthracnose pathogen of pepper and tomato, were developed using PCR. A specific primer set, coccoTef-F/coccoTef-R, which was constructed by analyzing tef-$1{\alpha}$ genes from 13 species and 22 strains of Colletotrichum, could specifically detect C. coccodes at a level of 10 ng by conventional PCR method and at 10 pg by real-time PCR. The PCR-based methods were also capable of detecting C. coccodes in pepper and tomato seeds artificially infected with the pathogen. The developed PCR methods can be applied for rapid and accurate inspection of C. coccodes in the seeds intended for export or import.