• Title, Summary, Keyword: Simple Sequence Repeats

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SSR-Primer Generator: A Tool for Finding Simple Sequence Repeats and Designing SSR-Primers

  • Hong, Chang-Pyo;Choi, Su-Ryun;Lim, Yong-Pyo
    • Genomics & Informatics
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    • v.9 no.4
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    • pp.189-193
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    • 2011
  • Simple sequence repeats (SSRs) are ubiquitous short tandem duplications found within eukaryotic genomes. Their length variability and abundance throughout the genome has led them to be widely used as molecular markers for crop-breeding programs, facilitating the use of marker-assisted selection as well as estimation of genetic population structure. Here, we report a software application, "SSR-Primer Generator " for SSR discovery, SSR-primer design, and homology-based search of in silico amplicons from a DNA sequence dataset. On submission of multiple FASTA-format DNA sequences, those analyses are batch processed in a Java runtime environment (JRE) platform, in a pipeline, and the resulting data are visualized in HTML tabular format. This application will be a useful tool for reducing the time and costs associated with the development and application of SSR markers.

Genetic Diversity and Population Structure of Spiraea prunifolia for. simpliciflora by Inter-Simple Sequence Repeats (조팝나무의 유전적 다양성과 집단구조 분석을 위한 ISSR 분석)

  • Huh, Man-Kyu
    • Journal of Life Science
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    • v.19 no.9
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    • pp.1183-1189
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    • 2009
  • 85 individual Spiraea prunifolia for. simpliciflora (Rosaceae) were sampled to examine the genetic diversity and population structure of S. prunifolia for. simpliciflora populations. Inter-simple sequence repeats (ISSR) produced 65 polymorphic loci and identified 78 ISSR genotypes. Three multilocus genotypes were shared by more than one plant within a population. Total genetic diversity values ($H_T$) and inter-locus variation in the within-population genetic diversity ($H_S$) were 0.293 and 0.183, respectively. On a per-locus basis, the proportion of total genetic variation due to differences among populations ($G_{ST}$) was 0.373. This indicated that about 37.3% of the total variation was among populations. ISSR markers are very effective in classifying natural population levels of S. prunifolia for. simpliciflora in Korea. In addition, insights into the relative gene diversity among and within populations of S. prunifolia for. simpliciflora would be useful in plant breeding and also for the development of strategies for ex situ conservation of plant genetic resources.

Genotyping of avian pathogenic Escherichia coli by DNA fragment analysis for the differences in simple sequence repeats

  • Han, Mi Na;Byeon, Hyeon Seop;Han, Seong Tae;Jang, Rae Hoon;Kim, Chang Seop;Choi, Seok Hwa
    • Korean Journal of Veterinary Service
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    • v.41 no.4
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    • pp.257-262
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    • 2018
  • Avian pathogenic E. coli (APEC) causes severe economic losses in the poultry farms, due to systemic infections leading to lethal colisepticemia. It causes a variety of diseases from air sac infection to systemic spread leading to septicemia. Secondary infection contains opportunistic infections due to immunosuppression disease. Collibacillosis causes the great problems in the poultry industry in Korea. Thus, it is necessary to identify and classify the characteristics of E. coli isolate of chicken origin to confirm the diversity of symptoms and whether they are transmitted among the farms. Fragment analysis is identify the difference in the number of Variable-Number Tandem-Repeats (VNTRs) for genotyping. VNTRs have repeating structure (Microsatellite, Short tandem repeats; STR, Simple sequence repeats; SSR) in the chromosome. This region can be used as a genetic marker because of its high mutation rate. And various lengths of the amplified DNA fragment cause the difference in the number of repetition of the DNA specific site. The number of repetition sequences indicates the separated size of fragments, so the each fragments can be distinguished by specific samples. The results of the sample show that there is no difference in six microsatellite loci (yjiD, aidB, molR_1, ftsZ, b1668, yibA). There are differences among the farms in relation of the number of repetitions of other six microsatellite loci (ycgW, yaiN, yiaB, mhpR, b0829, caiF). Four (ycgW, yiaB, b0829, caiF) of these six microsatellite loci show statistically significant differences (P<0.05). It means that the analysis using four microsatellite loci including ycgW, yiaB, b0829, and caiF can confirm among the farms. Five E. coli samples in one farm have same SSR repetition at all markers. But, there are significant differences from other farms at Four (ycgW, yiaB, b0829, caiF) microsatellite loci. These results emphasize again that the four microsatellite loci makes a difference in the amplified DNA fragments, enabling it to be used for E. coli genotyping.

Simple Sequence Repeat (SSR) and GC Distribution in the Arabidopsis thaliana Genome

  • Mortimer Jennifer C;Batley Jacqueline;Love Christopher G;Logan Erica;Edwards David
    • Journal of Plant Biotechnology
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    • v.7 no.1
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    • pp.17-25
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    • 2005
  • We have mined each of the five A. thaliana chromosomes for the presence of simple sequence repeats (SSRs) and developed custom perl scripts to examine their distribution and abundance in relation to genomic position, local G/C content and location within and around transcribed sequences. The distribution of repeats and G/C content with respect to genomic regions (exons, UTRs, introns, intergenic regions and proximity to expressed genes) are shown. SSRs show a non-random distribution across the genome and a strong association within and around transcribed sequences, while G/C density is associated specifically with the coding portions of transcribed sequences. SSR motif repeat number shows a high degree of variation for each SSR type and a high degree of motif sequence bias reflecting local genome sequence composition. PCR primers suitable for the amplification of identified SSRs have been designed where possible, and are available for further studies.

(CA/GT)n Simple Sequence Repeat DNA Polymorphism in Chlamydomonas reinhardtii (녹조류 Chlamydomonas reinhardtii의 (CA/GT)n Simple Sequence Repeat DNA 다형현상)

  • ;;Marvin W. FAWLEY
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.2
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    • pp.113-117
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    • 1997
  • Simple sequence repeats (SSR) are widely dispersed throughout eukaryotic genomes, highly polymorphic, and easily typed using polymerase chain reaction (PCR). The objective of this study was to determine the polymorphism of different Chlamydomonas reinhartdtii strains and to determine the mode of inheritance of the SSR locus in Chlamydomonas. A genomic DNA library of C. reinhardtii was constructed and screened with a radiolabeled $(AC)_{11}$ probe for the selection of (CA/GT)n repeat clone. Selected clone was seqeuenced, and PCR primer set flanking (CA/GT)n sequence was constructed. PCR was used to specifically amplify the SSR locus from multiple isolates of C. reinhardtii. The locus was polymorphic in some of the C. reinhardtii isolates. However, the locus was amplified only 4 of 6 isolates of C. reinhardtii, not in other 2 isolates of C. reinhardtii, suggesting that this locus is not extensively conserved. A simple Mendelian inheritance pattern was found, which showed 2:2 segregation in the tetrads resulting from a cross between C. reinhardtii and C. smithii. Our results suggest that this simple sequence repeat DNA polymorphism will be useful for identity testing, population studies, linkage analysis, and genome mapping in Chlamydomonas.

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Mendelian Inheritance of Inter-Simple Sequence Repeats Markers in Abies Koreans Wilson (구상나무에 있어서 Inter-Simple Sequence Repeats Marker의 유전양식(遺傳樣式))

  • Hong, Yong-Pyo;Cho, Kyung-Jin;Kim, Yong-Yul;Shin, Eun-Kyeong
    • Journal of Korean Society of Forest Science
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    • v.87 no.3
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    • pp.422-428
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    • 1998
  • Polymerase chain reaction(PCR)-based inter-simple sequence repeats(I-SSR) markers were analyzed in 48 megagametophytes of a single tree of Abies koreana $W_{ILS}$. Nineteen of the 35 primers, screened with 6 megagametophyte DNA and produced the clearest amplification products in the preliminary experiment, were used for PCR with 48 megagametophyte DNAs sampled from a single tree. On the basis of the chi-square test, a total of 51 amplicons, amplified by the 19 primers, were revealed to be segregated according to the Mendelian ratio(i.e., 1 : 1 segregation ratio) in the 48 megagametophytes at 5% significance level. Based on the linkage analysis, the observed 51 Mendelian loci turned out to be unlinked each other, which suggested that they are evenly distributed in the genome. However, majority of RAPD markers are known to belong to the independent linkage blocks, which frequently results in the amplification of RAPD markers from the restricted regions of the genome. Owing to the nature of even distribution of the 51 loci observed in this study, the I-SSR markers could give better resolution of estimating genetic diversity from the whole genome than RAPD markers. And I-SSR markers are also more suitable than RAPD markers for reconstructing phylogenetic relationship by a cladistic method which requires to fulfil the assumption of independent evolution of the different characters.

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Simple Sequence Repeat (SSR)-Based Gene Diversity in Burkholderia pseudomallei and Burkholderia mallei

  • Song, Han;Hwang, Junghyun;Myung, Jaehee;Seo, Hyoseok;Yi, Hyojeong;Sim, Hee-Sun;Kim, Bong-Su;Nierman, William C.;Kim, Heenam Stanley
    • Molecules and Cells
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    • v.27 no.2
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    • pp.237-241
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    • 2009
  • Pathogens Burkholderia pseudomallei (Bp) and Burkholderia mallei (Bm) contain a large number (> 12,000) of Simple Sequence Repeats (SSRs). To study the extent to which these features have contributed to the diversification of genes, we have conducted comparative studies with nineteen genomes of these bacteria. We found 210 genes with characteristic types of SSR variations. SSRs with nonamer repeat units were the most abundant, followed by hexamers and trimers. Amino acids with smaller and nonpolar R-groups are preferred to be encoded by the variant SSRs, perhaps due to their minimal impacts to protein functionality. A majority of these genes appears to code for surface or secreted proteins that may directly interact with the host factors during pathogenesis or other environmental factors. There also are others that encode diverse functions in the cytoplasm, and this protein variability may reflect an extensive involvement of phase variation in survival and adaptation of these pathogens.

Genome Research on Peach and Pear

  • Hayashi, Tateki;Yamamoto, Toshiya
    • Proceedings of the Korean Society of Plant Biotechnology Conference
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    • pp.101-109
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    • 2002
  • A lot of SSRs (simple sequence repeats) in peach and pear from enriched genomic libraries and in peach from a cDHA library were developed. These SSRs were applied to other related species, giving phenograms of 52 Prunus and 60 pear accessions. Apple SSRs could also be successfully used in Pyrus spp. Thirteen morphological traits were characterized on the basis of the linkage map obtained from an Fa population of peach. This map was compiled with those morphological markers and 83 DHA markers, including SSR markers used as anchor loci, to compare different peach maps. Molecular markers tightly linked to new root-knot nematode resistance genes were also found. A linkage map including disease-related genes, pear scab resistance and black spot susceptibility, in the Japanese pear Kinchaku were constructed using 118 RAPD markers. Another linkage map, of the European pear Bartlett, was also constructed with 226 markers, including 49 SSRs from pear, apple, peach and cherry. Maps of other Japanese pear cultivars, i.e., Kousui and Housui, were also constructed. These maps were the first results of pear species.

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Genome Research on Peach and Pear

  • Hayashi, Tateki;Yamamoto, Toshiya
    • Journal of Plant Biotechnology
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    • v.4 no.2
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    • pp.45-52
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    • 2002
  • A lot of SSRs (simple sequence repeats) in peach and pear from enriched genomic libraries and in peach from a cDNA library were developed. These SSRs were applied to other related species, giving phenograms of 52 Prunes and 60 pear accessions. Apple SSRs could also be successfully used in Pyrus spp. Thirteen morphological traits were characterized on the basis of the linkage map obtained from an $F_2$ population of peach. This map was compiled with those morphological markers and 83 DNA markers, including SSR markers used as anchor loci, to compare different peach maps. Molecular markers tightly linked to new root-knot nematode resistance genes were also found. A linkage map including disease related genes, pear scab resistance and black spot susceptibility, in the Japanese pear Kinchaku were constructed using 118 RAPD markers. Another linkage map, of the European pear Bartlett, was also constructed with 226 markers, including 49 SSRs from pear, apple, peach and cherry. Maps of other Japanese pear cultivars, i.e., Kousui and Housui, were also constructed. These maps were the first results of pear species.

Determination and Application of Combined Genotype of Simple Sequence Repeats (SSR) DNA Marker for Cultivars of Cymbidium goeringii (춘란(Cymbidium goeringii) 품종에 대한 Simple Sequence Repeats (SSR) DNA 마커의 복합 유전자형 결정과 적용)

  • Lee, Dae-Gun;Koh, Jae-Chul;Chung, Ki-Wha
    • Horticultural Science & Technology
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    • v.30 no.3
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    • pp.278-285
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    • 2012
  • Cymbidium goeringii is one of the most important and popular species in the orchid family in north-east Asia. In the present study, we prepared multiplex PCR system, and used it for the genotyping of eight simple sequence repeats (SSR) markers (CG409, CG415, CG709, CG722, CG787, CG1023, CG1210, and CG1281) in subject with 40 samples of cultivated varieties. All the analyzed samples showed different combined genotypes. The average combined power of discrimination was very high value of $7.14{\times}10^{-10}$, and observed heterozygosity (Ho = 0.466) was similar with two wild populations of C. goeringii, which may indicate no or little occurrence of genetic change after collection from wild habitats. The present study also developed a two-dimensional barcode to express information of genotype results of eight SSR markers (SSR DNA ID). The discrimination power of DNA ID between two individuals will be statistically more than 99.999999%. The SSR DNA ID and two-dimensional barcode may be very usefully applied for the discrimination and maintence of cultivars of C. goeringii.