• 제목/요약/키워드: PCR Polymorphism

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Isolation and Characterization of the Mutans Streptococci from the Dental Plaques in Koreans

  • Yoo, So-Young;Park, Seon-Joo;Jeong, Dong-Ki;Kim, Kwang-Won;Lim, Sung-Hoon;Lee, Sang-Ho;Choe, Son-Jin;Chang, Young-Hyo;Park, In-Soon;Kook, Joong-Ki
    • Journal of Microbiology
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    • 제45권3호
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    • pp.246-255
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    • 2007
  • Mutans streptococci have been implicated as cariogenic bacteria in dental caries because they can produce high levels of dental caries-causing lactic acid and extracellular polysaccharide. The aim of this study was to isolate and characterize the mutans streptococci from the dental plaque obtained from Koreans. The dental plaque samples were collected from the anterior and molar teeth of both jaws in 155 subjects (aged 2 to 33.2 years, average age $13.7{\pm}4.7\;years$). The samples were diluted by 100-fold in $1{\times}\;PBS$ and plated on mitis-salivarius bacitracin (MSB) agar plates. The mutans streptococci grown on MSB plates were screened by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) targeting dextranase gene (dex). The mutans streptococci were identified at the species level using a 16S rDNA sequencing comparison method. The biochemical tests were carried out to biotype the mutans streptococci. Ninety-five strains of the mutans streptococci out of 358 colonies, which were derived from 141 subjects, were isolated. Of them, 77 strains and 18 strains were Streptococcus mutans and Streptococcus sobrinus, respectively. The biotyping data showed that 62, 1, 20, 10, and 2 strains were biotypes I, II, IV, V and variant, respectively. Of the two strains of variant biotype, one strains was similar to biotype IV except that it was positive to the arginine hydrolysis test. We considered this one strain a new biotype, and classified it as biotype VII. In conclusion, S. mutans and its biotype I was most frequently isolated in Korean dental plaque. The mutans streptococci strains isolated in this study might be useful for the study of the pathogenesis and the prevention of dental caries.

The Genetic Approach on Analyzing the Habitat Characteristics of Fairy Pitta Pitta Nympha Inhabiting Jeju Island, the Korean Peninsula and Taiwan (제주도, 한반도 및 대만 내 팔색조의 생태적 서식특성 분석을 위한 유전적 접근)

  • Kim, Eun-Mi;Jeon, Yeon-Seon;Kim, Se-Jae;Kang, Chang-Wan;Won, Hyun-Kyu;Jeong, Gil-Sang
    • Journal of the Korean Society of Environmental Restoration Technology
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    • 제17권1호
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    • pp.81-90
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    • 2014
  • A Fairy Pitta is a bird known to breed only in mainland China, Taiwan, Japan and Korea and is listed as Vulnerable in the IUCN Red List. We carried out a DNA analysis to contribute to conserve the genetic diversity of Fairy Pitta. 32 samples were collected at Jeju Island, the Korean Peninsula and Taiwan from 2004 to 2013 and DNA was extracted from them and several sequences were amplified-it through PCR. And then we performed the population genetic analysis. We found there was a transversion between nucleotide sequences at CO1 gene, while there was no changes at Cyt-b gene. And we confirmed the polymorphism from two genes was caused from genetic drift not from selection. Through this analysis, the group within the Peninsula was found bigger than other two groups based on the analysis of CO1 gene, and the group from Taiwan was found bigger than other two groups through the analysis of Cyt-b gene. The population genetic structure of mitochondria gene of three group was showing CO1 gene had 5 haplotypes and Cyt-b gene had 6 haplotypes. Haplotype 2 in CO1 gene was found in three group and many individuals of samples had this haplotype. Like CO1 gene, haplotype 2 in Cyt-b gene was found in three group and was included in plenty of individuals. Other haplotypes were not overlaped and broke off among the three groups. To prevent from the extinction of Fairy Pitta and to obtain the genetic diversity, we need to compare with other regional group such as Japan, China and perform additional research in the non-breeding area.

Changes in enzyme activity and expression of DHFR of Toxoplumc gondii by antifolates (Antifolate 약제에 대한 톡소포자충의 DHFR 효소활성 및 유전자 발현의 변화)

  • 백은정;남호우
    • Parasites, Hosts and Diseases
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    • 제36권3호
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    • pp.191-198
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    • 1998
  • The responses to antifolales of ToxopLasmc Bondii were investigated by measuring the dihydrorolate redLlctase (DHFR) activity. quantity of DHFR mRNA, and single-strand conformational polymorphism (SSCP) pattern. Pyrimethamine (PYM) and methotrexate (MTX) were tested ds anlifolates. When T. gondii was treated wish PYM, the viability was decreased by the increasing concentration of PYM. DHFR activity tended to increase as the passage proceeded. and the quantity of mRNA expressed was also increased according to passages. The viability of 7. gonnii was decreased by the increasing concentration of MTX, but it was maintained over 40% up to $100{\;}{\mu\textrm{m}}$ MTX. DHFR activity was 77.4% in the 1st passage ($1{\;}{\mu\textrm{m}}$). 82.2% in the 4th passage ($10{\;}{\mu\textrm{m}}$), and 141.3% in the 7th passage ($100{\;}{\mu\textrm{m}}$) But no changes were detected in SSCP pattern of T gondii rxposvd to FYM and MTX. both. These results suggested that the response of T gondii to FYN was rofulalcd by transrriptional level and that, in MTX. the viability of T. gonnii was derived from increasing DHFK activity.

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Genetic Diversity of Rehmannia glutinosa Genotypes Assessed by Molecular Markers (분자표지자에 의한 지황 유전집단의 유전적 다양성)

  • Bang, Kyong-Hwan;Chung, Jong-Wook;Kim, Young-Chang;Lee, Jei-Wan;Kim, Hong-Sig;Kim, Dong-Hwi
    • Journal of Life Science
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    • 제18권4호
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    • pp.435-440
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    • 2008
  • Random amplified polymorphic DNA (RAPD) markers were used to identify the genetic diversities among and within varieties and landraces of Rehmannia glutinosa. Polymorphic and reproducible bands were produced by 10 primers out of total 20 primers used in the experiment. In RAPD analysis of the 11 genotypes, 64 fragments out of 73 amplified genomic DNA fragments were polymorphic which represented an average 6.4 polymorphic fragments per primer. Number of amplified fragments with random primers ranged from 2 (OPA-1) to 13 (OPA-11) and varied in size from 200 bp to 1,400 bp. Especially, OPA-10, OPA-11 and OPA-19 primers showed specific bands for varieties of Korea Jiwhang and Jiwhang il ho, which could be useful for discriminating from other varieties and landraces of R. glutinosa. Percentage polymorphism ranged from a minimum of 50% (OPA-1) to a maximum of 100% (OPA-11), with an average of 87.7%. Similarity coefficients were higher in the genotypes of Korea Jiwhang and Jiwhang il ho than in other populations. In cluster analysis, genotypes of Korea Jiwhang, Jiwhang il ho, and Japanese accession were separated from those of other varieties and landraces. Average of genetic diversity within the population $(H_S)$ was 0.110, while average of total genetic diversity $(H_T)$ was 0.229. Across all RAPD makers the $G_{ST}$ value was 0.517, indicating that about 52% of the total genetic variation could be explained by RAPDs differences while the remaining 48% might be attributable to differences among samples. Consequently, RAPD analysis was useful method to discriminate different populations such as domestic varieties and other landraces. The results of the present study will be used to understand the population and evolutionary genetics of R. gllutinosa.

Analysis of Genetic Relationship Among Collected Cymbidium goeringii Based on RAPD (RAPD를 이용한 춘란 수집종 20 품종의 유연관계 분석)

  • Kim, Tae Bok;Lee, Jin Jae;Song, Young Ju;Choi, Chang Hak;Cheong, Dong Chun;Yu, Young Jin
    • FLOWER RESEARCH JOURNAL
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    • 제19권4호
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    • pp.225-230
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    • 2011
  • This research was performed for making data-base of cross-breeding between Cymbidium goeringii cultivars. Morphological characteristics were investigated and then genetic relationship was analyzed. Collected 20 Cymbidium goeringii cultivars were clustered into 2 groups. Seven cultivars were clustered into group I, and thirteen cultivars were clustered into group II. Group I doesn't have leaf pattern. Group have leaf pattern. The genetic relationship among collected 20 Cymbidium goeringii cultivars was anaylzed using RAPD with ten 10-mers random primer. Eighty-nine bands were generated by RAPD. Among the rest, three bands were monomorphic and eight-six bands were polymorphic. Overall similarity degree ranged from 0.521 to 0.862. The result of RAPD analysis was clustered into 2 groups, too. Sixteen cultivars were clustered into GroupX, and four cultivars were clustered into GroupY. Result of classification with morphological characteristics and RAPD showed different pattern, but 4 cultivars of GroupY by RAPD analysis were included in groupby morphological characteristics. Crossbreeding combination among low related coltivars in RAPD analysis may get more efficient result.

Genomic Polymorphism Analysis Using Microsatellites in the Jeju Dogs (제주개의 microsatellite 마커를 이용한 유전적 다양성 분석)

  • Ko, Minjeong;Kwon, Seulgi;Kim, Hye-Ran;Byun, Jae-Hyun;Kim, Dae-Cheol;Choi, Bong-Hwan
    • Journal of Life Science
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    • 제29권6호
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    • pp.637-644
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    • 2019
  • This study was conducted to analyze the genetic characteristics of the Jeju dog for preservation and protection. A total of 139 dogs from 7 dog breeds, including the Jeju dog, were genotyped using 16 microsatellite markers. The results revealed 2-18 alleles per locus, with a total of 131 alleles among the 16 markers. Most alleles were identified for FH3381, which had 18 alleles, whereas FH2834 had the fewest alleles, with just 2. When the total mean value was observed, the expected heterozygosity and observed heterozygosity were higher for than for outgroup dogs, and the PIC values ranged from 0.000 to 0.862, respectively. The phylogenetic tree analysis of the Jeju dog and other dog varieties revealed that the Jeju dog is closest to the Sapsal dog (0.393). The phylogeny between the Jeju and Korean domestic dogs showed that the Jeju dog is most distant from the Dongkyung dog (0.507). Looking at the distribution individually, the Jeju dog is in the same group as the Labrador Retriever and the Sapsal dog. Meanwhile, the Poongsan, Dongkyung, and Jindo dogs and the German Shepherd were in the same group. Genetic information confirmed through the results of this study can be used as basic data to study the genetic characteristics of the Jeju dog.

Characterization of Phenotypic Traits and Application of Fruit Flesh Color Marker in Melon (Cucumis melo L.) Accessions (멜론 유전자원의 생육 평가와 과육색 유전형 분석)

  • Bae, Ik Hyun;Kang, Han Sol;Jeong, Woo Jin;Ryu, Jae Hwang;Lee, Oh Hum;Chung, Hee
    • Korean Journal of Plant Resources
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    • 제34권5호
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    • pp.478-490
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    • 2021
  • We aimed to generate basic breeding data for melon (Cucumis melo L.). A total of 219 melon accessions conserved at the National Agrobiodiversity Center (NAC) in Rural Development Administration (RDA) were used in this study, of which 72 (33%) were collected from India. The majority of accessions showed orange (42%) and white (36%) flesh color. In addition to phenotypic evaluations, the accessions were genotyped using a molecular marker for the carotenoid biosynthesis gene CmOr. DNA fragments of the expected size were amplified in 205 out of 219 accessions. Digestion of the PCR products with HinfI restriction endonuclease showed 100% concordance between phenotype and genotype in green-fleshed accessions, but 98%, 97%, and 80% concordance in orange-, white-, and creamy-fleshed accessions, respectively. Sequence analysis revealed single nucleotide changes in the three positions of SNP1, SNP2 and SNP1int in the CmOr gene among accessions. These newly found alleles suggest that there are multiple mechanisms in determining fruit flesh color in melon. Also, the phenotype data of diverse accessions obtained in this study will be a valuable source for melon breeding.

Polymorphism analysis of tri- and tetranucleotide repeat microsatellite markers in Hanwoo cattle

  • Shil Jin;Jeong Il Won;Hyoun Ju Kim;Byoungho Park;Sung Woo Kim;Ui Hyung Kim;Sung-Sik Kang;Hyun-Jeong Lee;Sung Jin Moon;Myung Sun Park;Yong Teak Sim;Sun Sik Jang;Nam Young Kim
    • Journal of Animal Science and Technology
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    • 제66권4호
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    • pp.717-725
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    • 2024
  • The Hanwoo traceability system currently utilizes 11 dinucleotide repeat microsatellite (MS) markers. However, dinucleotide repeat markers are known to have a high incidence of polymerase chain reaction (PCR) artifacts, such as stutter bands, which can complicate the accurate reading of alleles. In this study, we examined the polymorphisms of the 11 dinucleotide repeat MS markers currently employed in traceability systems. Additionally, we explored four trinucleotide repeat MS markers and one tetranucleotide repeat MS marker in a sample of 1,106 Hanwoo cattle. We also assessed the potential utility of the tri- and tetranucleotide repeat MS markers. The polymorphic information content (PIC) of the five tri- and tetranucleotide repeat markers ranged from 0.663 to 0.767 (mean: 0.722), sufficiently polymorphic and slightly higher than the mean (0.716) of the current 11 dinucleotide repeat markers. Using all 16 markers, the mean PIC was 0.718. The estimated probability of identity (PI) was 3.13 × 10-12 using the 11 dinucleotide repeat markers, 7.03 × 10-6 using the five tri- and tetranucleotide repeat markers, and 2.39 × 10-17 using all 16 markers; the respective PIhalf-sibs values were 2.69 × 10-9, 1.29 × 10-4, and 3.42 × 10-13; and the respective PIsibs values were 3.89 × 10-5, 9.6 × 10-3, and 3.69 × 10-7. The probability of exclusion1 (PE1) was 0.999864 for the 11 dinucleotide repeat markers, 0.981141 for five of the tri- and tetranucleotide repeat markers, and > 0.99 for all 16 markers; the respective PE2 values were 0.994632, 0.901369, and > 0.99; and the respective PE3 values were 0.998702, > 0.99, and > 0.99. The five investigated triand tetranucleotide repeat MS markers can be used in combination with the 11 existing MS markers to improve the accuracy of individual identification and paternity testing in Hanwoo.

Development and Validation of an SNP Marker for Identifying Xanthomonas oryzae pv. oryzae Thai Isolates That Break xa5-Mediated Bacterial Blight Resistance in Rice

  • Tebogo Balone;Ananda Nuryadi Pratama;Werapat Chansongkram;Thanita Boonsrangsom;Kawee Sujipuli;Kumrop Ratanasut
    • The Plant Pathology Journal
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    • 제40권5호
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    • pp.451-462
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    • 2024
  • Xanthomonas oryzae pv. oryzae (Xoo) is a pathogenic bacterium responsible for bacterial blight (BB) disease in rice, primarily mediated by the interaction between the plant and pathogen. The virulence mechanism involves the activation of the Sugars Will Eventually be Exported Transporter (SWEET) gene family in rice by transcription activator-like effectors derived from Xoo. The BB resistance gene xa5 has been identified as one of the most effective genes against Thai Xoo isolates, but xa5-mediated resistance-breaking Xoo strains have emerged. This study aimed to develop a single nucleotide polymorphism (SNP) marker for precise identification of xa5-mediated resistance-breaking Xoo. Comparative genomics of Thai Xoo isolates Xoo16PK001 and Xoo16PK002, which were incompatible and compatible with rice variety IRBB5 carrying xa5, respectively, identified eight SNP positions for the development of an SNP marker. The SNP marker XooE6 yields a specific 1,143 bp PCR product unique to Xoo16PK002. Screening 61 Thai isolates using XooE6 identified two positives: Xoo20PL010 and Xoo20UT002. Inoculation tests on rice varieties IRBB5 and IRBB13 demonstrated compatibility with IRBB5 and incompatibility with IRBB13, which bears Xa5 and xa13. Xoo16PK001 (XooE6-negative) showed different virulence. Inoculation on IRBB21 harboring Xa5, Xa13, and Xa21 resulted in partial resistance to both XooE6-positive and -negative strains. XooE6-positive strains up-regulated SWEET11 and suppressed SWEET14 in IRBB5, while Xoo16PK001 slightly induced SWEET11 but activated SWEET14 in IRBB13. This highlights the potential of XooE6 to identify xa5-mediated resistance-breaking Xoo strains and elucidate their pathogenic mechanisms through the upregulation of SWEET11.

Characterization of the DGAT1 Gene in the Korean Holstein Dairy Cattle Population (한국 Holstein종 유우집단의 DGAT1 유전자의 특성분석)

  • Son, Ji-Young;Jeong, Hang-Jin;Yu, Seong-Lan;Lee, Jun-Heon;Do, Chang-Hee;Ryoo, Seung-Heui;Sang, Byung-Chan
    • Korean Journal of Agricultural Science
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    • 제36권2호
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    • pp.167-177
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    • 2009
  • This study was conducted to characterize the DGAT1 gene in Korean Holstein dairy cattle population and examine the relationship of DGAT1 polymorphisms with milk yield and milk fat yield for the genetic improvement of Korean Holstein dairy cattle. Results indicated that the 411 bp PCR products were successfully amplified by DGAT1 specific primers. Sequence analysis indicated that the DGTA1 Q allele had AUG (Lysine, K) nucleotide sequences in 216-218 bp and q allele had GCG (Alanine, A) sequences in the same position. Nucleotide sequence homology between the DGAT1 sequences generated in this study showed 100% homology with bovine DGAT1 sequences in the NCBI database. The genotype frequencies of DGAT1 QQ, Qq, and qq were 16.43, 36.43, and 47.14%, respectively, in Korean Holstein dairy cattle population. The observed Q and q allele frequencies were 0.35 and 0.65, respectively. Statistically significant (P<0.05) results were identified for milk yield and milk fat yield for the DGAT1 genotypes. The Qq genotype Holsteins have significantly higher milk yield and milk fat yield than those of the QQ and qq genotype Holsteins(P<0.05).

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