• Title/Summary/Keyword: Simple Sequence Repeats

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Identification of Non-Muscle Nebulin Isoform in Human Brain Library

  • Joo, Young-Mi;Lee, Min-A;Choi, Pyung-Rak;Choi, Jae-Kyoung;Lee, Yeong-Mi;Choi, Su-Il;Kim, Myong-Shin;Jeon, Eun-Hee;Kim, So-Young;Kim, Chong-Rak
    • Biomedical Science Letters
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    • v.10 no.1
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    • pp.23-29
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    • 2004
  • Nebulin is a (Mr 600∼900 kDa) large actin-binding protein specific to skeletal muscle and thought to act as a molecular template that regulates the length of thin filaments. Cardiac muscles of higher vertebrates have been shown earlier to lack nebulin. Recently, full-length nebulin mRNA transcripts have been detected in heart muscle, but at lower levels than in skeletal muscle. Nebulin expression also was detected in the kidney, eye, and otic canal, suggesting that nebulin isoforms may also be expressed in these organs. We have searched for nebulin isoforms in brain of human using PCR and Northern blot. Here, we provide evidence that nebulin mRNA transcripts are expressed in brain. Seven nebulin isoforms (B, C, D, E, F, G and H form) are obtained in human skeletal muscle and four isoforms (B, C, G and H form) in human brain cDNA library. We cloned the 1.3 kb of nebulin fragment from human adult brain library by PCR. The identity of the PCR product was confirmed by sequence analysis. The partial brain nebulin sequence was 99% identical to the skeletal muscle cDNA as determined by Blast alignment. It contains two simple-repeats HR1, HR2 and linker-repeats exon l35∼143 except exon 140. It was different from skeletal muscle B form, which contain HR1 and HR8. These data suggest that nebulin isoform diversity occurs even more extensively than previously known, likely contributing to the distinct thin filament architecture of different striated muscles.

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Marker Production by PCR Amplification with Primer Pairs from Conserved Sequences of WRKY Genes in Chili Pepper

  • Kim, Hyoun-Joung;Lee, Heung-Ryul;Han, Jung-Heon;Yeom, Seon-In;Harn, Chee-Hark;Kim, Byung-Dong
    • Molecules and Cells
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    • v.25 no.2
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    • pp.196-204
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    • 2008
  • Despite increasing awareness of the importance of WRKY genes in plant defense signaling, the locations of these genes in the Capsicum genome have not been established. To develop WRKY-based markers, primer sequences were deduced from the conserved sequences of the DNA binding motif within the WRKY domains of tomato and pepper genes. These primers were derived from upstream and downstream parts of the conserved sequences of the three WRKY groups. Six primer combinations of each WRKY group were tested for polymorphisms between the mapping parents, C. annuum 'CM334' and C. annuum 'Chilsung-cho'. DNA fragments amplified by primer pairs deduced from WRKY Group II genes revealed high levels of polymorphism. Using 32 primer pairs to amplify upstream and downstream parts of the WRKY domain of WRKY group II genes, 60 polymorphic bands were detected. Polymorphisms were not detected with primer pairs from downstream parts of WRKY group II genes. Half of these primers were subjected to $F_2$ genotyping to construct a linkage map. Thirty of 41 markers were located evenly spaced on 20 of the 28 linkage groups, without clustering. This linkage map also consisted of 199 AFLP and 26 SSR markers. This WRKY-based marker system is a rapid and simple method for generating sequence-specific markers for plant gene families.

A Survey of the Brassica rapa Genome by BAC-End Sequence Analysis and Comparison with Arabidopsis thaliana

  • Hong, Chang Pyo;Plaha, Prikshit;Koo, Dal-Hoe;Yang, Tae-Jin;Choi, Su Ryun;Lee, Young Ki;Uhm, Taesik;Bang, Jae-Wook;Edwards, David;Bancroft, Ian;Park, Beom-Seok;Lee, Jungho;Lim, Yong Pyo
    • Molecules and Cells
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    • v.22 no.3
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    • pp.300-307
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    • 2006
  • Brassica rapa ssp. pekinensis (Chinese cabbage) is an economically important crop and a model plant for studies on polyploidization and phenotypic evolution. To gain an insight into the structure of the B. rapa genome we analyzed 12,017 BAC-end sequences for the presence of transposable elements (TEs), SSRs, centromeric satellite repeats and genes, and similarity to the closely related genome of Arabidopsis thaliana. TEs were estimated to occupy 14% of the genome, with 12.3% of the genome represented by retrotransposons. It was estimated that the B. rapa genome contains 43,000 genes, 1.6 times greater than the genome of A. thaliana. A number of centromeric satellite sequences, representing variations of a 176-bp consensus sequence, were identified. This sequence has undergone rapid evolution within the B. rapa genome and has diverged among the related species of Brassicaceae. A study of SSRs demonstrated a non-random distribution with a greater abundance within predicted intergenic regions. Our results provide an initial characterization of the genome of B. rapa and provide the basis for detailed analysis through whole-genome sequencing.

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 Microsatellite Markers to Distinguish South Korean and Chinese Ginseng

  • Ahn, Chang-Ho;Kim, Boo-Bae;Yoon, Eui-Soo;Choi, Yong-Eui
    • Journal of Korean Society of Forest Science
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    • v.98 no.5
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    • pp.568-575
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    • 2009
  • Korean wild and forest cultivated ginseng has long been accepted as high medicinal values compared to field cultivated ginseng. Owing to the high price of Korean wild ginseng, Chinese wild and forest cultivated ginseng were smuggled and sold as Korean wild and forest cultivated ginseng. Therefore, an efficient method is required to distinguish Korean ginseng from Chinese ginseng. Microsatellites, simple sequence repeats (SSRs), are highly polymorphic loci present in DNA that consist of repeating units of base pairs. Thus SSR markers are highly advantageous for detection of small genetic variances of intra-species. In the present study, we constructed a microsatellite-enriched genomic library from South Korean wild Panax ginseng. After sequence analysis of 992 randomly picked positive colonies, 126 (12.7%) of the colonies were found to contain microsatellite sequences, and 38 primer pairs were designed. By polymorphism assessment using 36 primer pairs, 4 primers (PG409, PG450, PG491, and PG582) were shown to be polymorphic to distinguish the South Korean ginseng from the Chinese ginseng. These 4 microsatellite markers will provide powerful tools to authenticate South Korean ginseng from Chinese ginseng.

Genetic variation of the endangered species Halenia coreana (Gentianaceae)

  • YUN, Narae;OH, Sang-Hun
    • Korean Journal of Plant Taxonomy
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    • v.52 no.1
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    • pp.45-53
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    • 2022
  • Halenia coreana is an endangered, endemic species that is distributed in only a few locations in Korea, such as Mts. Hwaaksan and Daeamsan. It has been recently segregated from H. corniculata, broadly distributed in cold temperate regions that include northern Japan, the Russian Far East, northeastern China, Mongolia, and eastern Europe, where population sizes are usually large. To examine the genetic diversity of H. coreana and evaluate the level of genetic differentiation of the species compared with that of H. corniculata, we surveyed 183 candidate simple sequence repeats (SSR) motif markers for H. coreana and H. corniculata from sequence data of amplified fragments of a specific length in the genome. A total of 17 genomic-SSR markers were selected to examine the levels of genetic diversity and differentiation using 17 samples of H. coreana and 60 samples of three populations of H. corniculata. The results here suggest that the genetic diversity of H. coreana is very low with a high frequency of inbreeding within its population. We found that H. coreana is genetically differentiated from H. corniculata, supporting the recognition of the geographically isolated H. coreana as a distinct species.

Genetic Diversity of High-Quality Rice Cultivars Based on SSR Markers Linked to Blast Resistance Genes (도열병 저항성 유전자와 연관된 SSR 마커를 이용한 양질미 품종의 유전적 다양성)

  • Huhn-Pal Moon
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.49 no.3
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    • pp.251-255
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    • 2004
  • The epidemics of rice blast which occurred in south parts of Korea during the period from 1999 to 2001 and damaged several high quality rice cultivars developed using "Milyang 95" and/or "Milyang 96" as a parent. Genetic diversity of 23 rice cultivars including "Milyang 95" and it's relatives was assessed using 54 simple sequence repeats (SSR) markers reported to be linked to major blast resistance genes. Fifty-four SSR markers representing fifty-seven loci in the rice genome detected polymorphism among the 23 cultivars and revealed a total of 170 alleles with an average of 3.0 alleles per primer, The number of amplified bands ranged from 1 to 7. Several SSR markers including RM249, RM206 and OSR20 were informative for assessing the genetic diversity of relatively closed japonica rice cultivars. The 23 cultivars were classified into four groups by cluster analysis based on Nei's genetic distances, and the cultivars developed from same parents showed a tendency to cluster together that is consistant with genealogical information. High quality rice cultivars, Daesanbyeo, Donganbyeo, and Milyang 95 belonged to the same cluster, At the loci, RM254 and OSR32, all of the cultivars derived from the crosses using "Milyang 95" shared same alleles, suggesting that these japonica cultivars might carry alleles that are identical by descent. Evaluation of 23 rice cultivars against blast needs to be confirmed regarding the relationship between genotype and blast resistance.p between genotype and blast resistance.

Genetic diversity of Forsythia ovata Nakai (Oleaceae) based on inter-simple sequence repeats (ISSR) (ISSR 자료에 기초한 만리화(물푸레나무과)의 유전적 다양성)

  • Kim, Sang-Yong;Kim, Young-Dong;Kim, Jin-Seok;Yang, Byeong-Hoon;Kim, Sung-Hee;Lee, Byung-Chun
    • Korean Journal of Plant Taxonomy
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    • v.39 no.1
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    • pp.48-54
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    • 2009
  • We investigated the genetic diversity of an endemic rare species, Forsythia ovata Nakai by examining 93 ISSR amplicons in 84 individuals distributed among five populations. The overall percentage of polymorphic ISSR amplicons was 54.8% and mean number of amplicons per ISSR primer was 6.6. The amount of genetic diversity was relatively lower than other shrub species. The Mt. Seokbyeong and Mt. Seorak B populations had the highest level of genetic diversity. Although the Seokgae-jae population had the lowest level of genetic diversity, the population was genetically the most distinctive from the other populations. About 30.6% of the total variation was allocated between five populations, which was slightly higher than other shrub species. Such a pattern of genetic variation may have resulted from the limited distribution and small population sizes of F. ovata. The UPGMA dendrogram based on Nei's genetic distance showed some decisive geographic patterns. These results suggest that, in addition to the preservation of the natural stands, the conservation of larger number of populations with small number of individuals per population is more effective for the dynamic ex situ conservation and for maintaining the genetic diversity of F. ovata than smaller number of populations with large number of individuals.

Development of Novel Microsatellite Markers for Strain-Specific Identification of Chlorella vulgaris

  • Jo, Beom-Ho;Lee, Chang Soo;Song, Hae-Ryong;Lee, Hyung-Gwan;Oh, Hee-Mock
    • Journal of Microbiology and Biotechnology
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    • v.24 no.9
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    • pp.1189-1195
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    • 2014
  • A strain-specific identification method is required to secure Chlorella strains with useful genetic traits, such as a fast growth rate or high lipid productivity, for application in biofuels, functional foods, and pharmaceuticals. Microsatellite markers based on simple sequence repeats can be a useful tool for this purpose. Therefore, this study developed five novel microsatellite markers (mChl-001, mChl-002, mChl-005, mChl-011, and mChl-012) using specific loci along the chloroplast genome of Chlorella vulgaris. The microsatellite markers were characterized based on their allelic diversities among nine strains of C. vulgaris with the same 18S rRNA sequence similarity. Each microsatellite marker exhibited 2~5 polymorphic allele types, and their combinations allowed discrimination between seven of the C. vulgaris strains. The two remaining strains were distinguished using one specific interspace region between the mChl-001 and mChl-005 loci, which was composed of about 27 single nucleotide polymorphisms, 13~15 specific sequence sites, and (T)n repeat sites. Thus, the polymorphic combination of the five microsatellite markers and one specific locus facilitated a clear distinction of C. vulgaris at the strain level, suggesting that the proposed microsatellite marker system can be useful for the accurate identification and classification of C. vulgaris.

Molecular Authentication and Genetic Polymorphism of Korean Ginseng (Panax ginseng C. A. Meyer) by Inter-Simple Sequence Repeats (ISSRs) Markers (ISSRs 마크에 의한 고려 인삼의 분자적 인증과 유전적 다형현상)

  • Bang, Kyong-Hwan;Lee, Sung-Woo;Hyun, Dong-Yun;Cho, Joon-Hyeong;Cha, Seon-Woo;Seong, Nak-Sul;Huh, Man-Kyu
    • Journal of Life Science
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    • v.14 no.3
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    • pp.425-428
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    • 2004
  • Molecular authentication and genetic polymorphism of Korean ginseng cultivars and accessions were investigated using ISSR (inter-simple sequence repeat amplification) markers. Five primers among 56 produced clear and reproducible DNA fragments among seven cultivars and accessions. A total of 43 bands ranging from 250 bp to 1,700 bp from five primers were scored. Average number of bands per primer was 8.6 and only nine bands were polymorphic across the six Panax ginseng from Korea. Especially Chunpoong cultivar exhibited the highest level of polymorphism, whereas other accessions did not showed almost any polymorphism. Consequently, these ISSR markers will be available to differentiate Chunpoong cultivar from other major Korean ginseng cultivars and accessions, such as Yunpoong, Hwangsukjong and Jakyungjong, at the DNA level.