• Title/Summary/Keyword: Sequence polymorphisms

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Discrimination of Korean Agaricus bisporus cultivars using CAPS markers (CAPS 마커를 이용한 국내 개발 양송이 품종 구분)

  • Lee, Hwa-Yong;An, Hyejin;Oh, Youn-Lee;Jang, Kab-Yeul;Chung, Jong-Wook
    • Journal of Mushroom
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    • v.19 no.4
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    • pp.336-340
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    • 2021
  • The cleaved amplified polymorphic sequence (CAPS) marker uses a restriction enzyme recognition site resulting from single nucleotide polymorphisms and insertions and deletions on the DNA sequence. This technique does not require expensive equipment, the process is simple, and clear results can be obtained reliably. In this study, Agaricus bisporus cultivars SaeA, SaeDo, SaeHan, SaeYeon, SaeJeong, Dodam, Seolgang, Dahyang, Hogam, and Hadam developed in Korea were discriminated using four CAPS markers. Our results indicated that it is possible to distinguish the ten cultivars and determine the genetic diversity among them.

Polymorphisms of HLA-DRB1 and -DQB1 Genes in Korean Patients with Pulmonary Tuberculosis (한국인 폐결핵 환자에서 HLA-DRB1 및 -DQB1 유전자의 다형성에 관한 연구)

  • Park, Myoung Hee;Song, Eun Young;Kwon, Sung Youn;Park, He Jin;Han, Sung Koo;Shim, Young Soo
    • Tuberculosis and Respiratory Diseases
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    • v.54 no.4
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    • pp.367-377
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    • 2003
  • Background : It is well known that only 10% of those infected with Mycobacterium tuberculosis actually develop clinical disease, indicating the existence of host genetic factors regulating disease expression. In this study, we investigated HLA-DRB1 and -DQB1 gene polymorphisms in Korean patients with pulmonary tuberculosis (PTB). Methods : HLA-DRB1 and -DQB1 gene polymorphisms were investigated in 67 PTB patients without previous treatment history, 38 drug-sensitive (DS) and 29 multidrug-resistant (MDR) cases, and 200 healthy controls. HLA-DRB1 typing was done using reverse SSO (sequence specific oligonucleotide) and PCR-SSCP (single strand conformational polymorphism) methods and DQB1 typing was done using PCR-RFLP (restriction fragment length polymorphism), PCR-SSCP and PCR-SSP (sequence specific primer) methods. Results : Among the PTB patients, MDR-TB cases showed frequencies of DRB1*0701 and *08032 increased by about two-fold compared to those of normal controls, and likewise for their associated DQB1 alleles, DQB1*0202 and *0601 (15.5% vs. 34.5%, p=0.01). The frequency of HLA-DQB1*0609 was significantly increased in PTB patients (4.0% vs. 14.9%, p=0.004), showing similar increases in both DS and MDR cases. There was also an association of HLA alleles with the clinical severity of the disease according to the extent of lung lesion. Significantly increased frequencies of DRB1*08032 (4.2% vs. 32.6%, p=0.007) and DQB1*0601 (12.5% vs. 34.9%, p=0.047) were observed in more advanced (moderately & far advanced/DS and far advanced/MDR), compared with less advanced (minimal/DS and moderately advanced/MDR) lung lesions. Although DRB1*0701, DQB1*0202 and DQB1*0609 showed significant increases in different subsets of the disease, these HLA alleles did not show consistent association with disease severity. Conclusion : HLA-DRB1*08032 and DQB1*0601 alleles were associated with genetic susceptibility to MDR-TB in Korean patients, and also with disease severity and progression of PTB.

Evidence of genome duplication revealed by sequence analysis of multi-loci expressed sequence tagesimple sequence repeat bands in Panax ginseng Meyer

  • Kim, Nam-Hoon;Choi, Hong-Il;Kim, Kyung Hee;Jang, Woojong;Yang, Tae-Jin
    • Journal of Ginseng Research
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    • v.38 no.2
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    • pp.130-135
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    • 2014
  • Background: Panax ginseng, the most famous medicinal herb, has a highly duplicated genome structure. However, the genome duplication of P. ginseng has not been characterized at the sequence level. Multiple band patterns have been consistently observed during the development of DNA markers using unique sequences in P. ginseng. Methods: We compared the sequences of multiple bands derived from unique expressed sequence tagsimple sequence repeat (EST-SSR) markers to investigate the sequence level genome duplication. Results: Reamplification and sequencing of the individual bands revealed that, for each marker, two bands around the expected size were genuine amplicons derived from two paralogous loci. In each case, one of the two bands was polymorphic, showing different allelic forms among nine ginseng cultivars, whereas the other band was usually monomorphic. Sequences derived from the two loci showed a high similarity, including the same primer-binding site, but each locus could be distinguished based on SSR number variations and additional single nucleotide polymorphisms (SNPs) or InDels. A locus-specific marker designed from the SNP site between the paralogous loci produced a single band that also showed clear polymorphism among ginseng cultivars. Conclusion: Our data imply that the recent genome duplication has resulted in two highly similar paralogous regions in the ginseng genome. The two paralogous sequences could be differentiated by large SSR number variations and one or two additional SNPs or InDels in every 100 bp of genic region, which can serve as a reliable identifier for each locus.

PCR-DGGE and PCR-RFLP Analyses of the Internal Trascribed Spacer(ITS) of Ribosomal DNA in the Genus Rhizopus

  • Park, You-jung;Park, Young-Keel;Min, Byung-Re
    • Journal of Microbiology
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    • v.41 no.2
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    • pp.157-160
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    • 2003
  • To estimate genetic relationships within the genus Rhizopus, genetic variations in 20 strains were investigated by DGGE and PCR-RFLP of rDNA ITS region (ITSI, ITS2,5.8S). The size of the amplified products showed the interspecific polymorphisms, 650 bp,700 bp, and 900 bp. The DGGE approach allowed the separation of PCR amplicons of the same length according to their sequence variations. When the rDNA ITS region was digested with six restriction enzymes, 20 strains were classified into five RFLP haplotypes. The range of similarity between the 20 strains by PCR-RFLP was 42.3-100%. Based on the results of DGGE aud PCR-RFLP, the 20 strains were divided into four groups, R. oryzae, R. stolonifer, R. microsporus and R. homothallicus. Further study of R. homothallicus is required.

The First Case of Korean Boy with Mitochondrial Trifunctional Protein Deficiency Diagnosed by Acylcarnitine Profiles and DNA analysis : A Novel Mutation in the α-subunit of the Mitochondrial Trifunctional Protein and a Unusual Intergenic Sequence with Two Polymorphisms

  • Lee, Ji-Eun;Yoon, Hye-Ran;Paik, Kyung Hoon;Hwang, Jong Hee;Hwang, Soo-Jung;Shim, Jae-Won;Chang, Yun-Sil;Park, Won-Soon;Strauss, Arnold W.;Jin, Dong-Kyu
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.2 no.1
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    • pp.12-14
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    • 2002
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Potential Applications of Recombinant DNA Probes for Relatedness Analysis of Fusarium oxysporum (Fuarium oxysporum의 유연관계 분석을 위한 Recombinant DNA의 Probe로서의 이용 가능성)

  • 김홍기;김영태;유승헌
    • Korean Journal Plant Pathology
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    • v.10 no.1
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    • pp.1-6
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    • 1994
  • Randomly chosen recombinant clones of Fusarium oxysporum were analysed to select useful probes for relatedness analysis of Fusarium oxysporum. Genomic DNA of F. oxysproum f. sp. cubense, digested with HindIII, was ligated to pUC118 and used to transform Escherichia coli strai DH5$\alpha$. Three clones were identified that hybridized to mutiple restriction fragments of some formae speciales of F. oxysporum. These probes detected repetitive sequences in HindIII or EcoRI digested DNAs. Repeated copy clone pFC46, pFC52 and pFC54 showed evident polymorphisms among ten formae speciales of this fungus. Since clone pFC 52 strongly hybridized to multiple EcoRI-digested restriction fragments of f. sp. cubense, it may be useful as a probe for analysis of other genetic characteristics of this forma specialis. The results suggest that our clones might be very useful as probes for relatedness analysis between or within formae speciales of Fusarium oxysporum.

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Improved characterization of Clematis based on new chloroplast microsatellite markers and nuclear ITS sequences

  • Liu, Zhigao;Korpelainen, Helena
    • Horticulture, Environment, and Biotechnology : HEB
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    • v.59 no.6
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    • pp.889-897
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    • 2018
  • Currently, there is a lack of genetic markers capable of effectively detecting polymorphisms in Clematis. Therefore, we developed new markers to investigate inter- and intraspecific diversity in Clematis. Based on the complete chloroplast genome of Clematis terniflora, simple sequence repeats were explored and primer pairs were designed for all ten adequate repeat regions (cpSSRs), which were tested in 43 individuals of 11 Clematis species. In addition, the nuclear ITS region was sequenced in 11 Clematis species. Seven cpSSR loci were found to be polymorphic in the genus and serve as markers that can distinguish different species and be used in different genetic analyses, including cultivar identification to assist the breeding of new ornamental cultivars.

Development of Restriction Fragment Length Polymorphism(RELP) Markers in Silkworm, Bombyx mori (누에 RFLP(제한단편 다형현상)마커 개발)

  • 고승주;김태산;이영승;황재삼;이상몽
    • Korean journal of applied entomology
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    • v.36 no.1
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    • pp.96-104
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    • 1997
  • A silkworm Bombyx mori genomic DNA library was constructed from polyphagous J111 strain and unpolyphagous $C_3$ strain to develop the genomic study by DNA makers. Genomic DNAs of two strains were digested with restriction enzyme EcoRI and ligated into pUC18. The ligated plasmids were transferred into E. coli host strain DH5$\alpha$. When the genomic DNAs were hybridized with insert DNAs from transformant, could be categorized from hybridization patterns to three groups as high repetitive sequence, moderately repetitive sequence, and low-copy number sequences. A total of 219 clones containing single or low-copy number sequence inserts were examined for any polymorphisms between two strains of J111 and $C_3$. Forty six clones showed RFLPs and 10 of these clones were used as a probe of analysis of $F_2$ population derived from crossing between J111 and $C_3$ strain. The genetic inheritance tested with each clones will be important tools to construct the genetic map of the silkworm, Bombyx mori.

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Full Length cDNA, Genomic Organizations and Expression Profiles of the Porcine Proteasomal ATPases PSMC5 Gene

  • Wang, Y.F.;Yu, M.;Liu, B.;Fan, B.;Wang, H.;Zhu, M.J.;Li, K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.7
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    • pp.897-902
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    • 2004
  • PSMC5 subunit, which belongs to the 26S proteasomal subunit family, plays an important role in the antigen presentation mediated by MHC class I molecular. Full-length cDNA of porcine PSMC5 was isolated using the in silico cloning and rapid amplification of cDNA ends (RACE). Amino acid was deduced and the primary structure was analyzed. Results revealed that the porcine PSMC5 gene shares the high degree of sequence similarity with its mammalian counterparts at both the nucleotide level and the amino acid level. The RT-PCR was performed to detect the porcine PSMC5 expression pattern in seven tissues and the result showed that high express level was observed in spleen, lung, marrow and liver while the low express level was in muscle. The full-length genomic DNA sequence of porcine PSMC5 gene was amplified by PCR and the genomic structure revealed that this gene was comprised by 12 exons and 11 introns. Best alignment of the cDNA and genomic exon DNA sequence presents 4 mismatches and this information potentially bears further study in gene polymorphisms.

Evaluation of Genetic Diversity among Soybean Genotypes Using SSR and SNP

  • Lee, Suk-Ha;P. Tanya;O, Srinives;T. Toojinda;A. Vanavichit;Ha, Bo-Keun;Bae, Jeong-Suk;Moon, Jung-Kyung
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.46 no.4
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    • pp.334-340
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    • 2001
  • Two different types of molecular markers, simple sequence repeat (SSR) and single nucleotide polymorphism (SNP), were used to measure genetic diversity among five Korean, eight Thai, and three wild soybeans. For SSR analysis, a total of 20 markers were surveyed to detect polymorphisms. For SNP analysis, four primers were designed from consensus sequence regions on disease resistance protein homolog genes, and used to amplify the genomic region. The PCR products were sequenced. A number of polymorphic SSR and SNP bands were scored on all genotypes and their genetic similarity was measured. Clustering analysis was performed independently on both types of markers. Clustering based on SSR markers separated the genotypes into three main groups originated from Korea, Thailand, and wild soybeans. On the other hand, two main groups were classified using SNP analysis. It seemed that SSR was more informative than SNP in this study. This may be due to the fact that SNP was surveyed on the smaller genomic region than SSR. Grouping based on the combined data of both markers revealed similar results to that of SNP rather than that of SSR. This might be due to the fact that more loci from SNP were considered to measure genetic relatedness than those from the SSR.

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