• Title/Summary/Keyword: primers

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Construction of a linkage Map in Capsicum annuum L. Using RAPD Markers and Identification of Two QTLs.

  • Yang, Tae-Jin;Kim, Yong-Jae;Park, Hyo-Guen
    • Journal of Plant Biotechnology
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    • v.1 no.2
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    • pp.109-115
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    • 1999
  • A linkage map of Capsicum annuum L. was constructed by random amplified polymorphic DNA (RAPD) markers followed in a backcross population of an intraspecific cross between cultivars HDA210 and Yatsufusa. A total of 420 random primers were tested and 311 polymorphic bands were generated by 158 random primers. Among them, 86 Yatsufusa specific bands generated by 52 primers were examined for mapping. Most bands except three segregated in Mendelian fashion fitting the expected 1:1 ratio. The total length of the map was 533 cM distributed in 15 linkage groups. The map distance between adjacent markers ranged 0 to 32.8 cM, with an average distance of 9.1 cM (63 markers). Some markers were clustered and this may be due to the amplification of a repetitive sequence by the RAPDs. Primer pairs for a sequence characterized amplified region (SCAR) were developed and the segregation scores by the SCAR primers were in accordance with the RAPD data. Two QTL markers for number of axillary shoots and for early flowering were developed. One QTL for early flowering located in the linkage group 3 and explained 61 "io of the phenotypic variation. The other QTL for the number of axillary shoots located in the linkage group 4 explained 55 % of the phenotypic variation.tion.

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Application of RAPD markers for characterization of ${\gamma}$-ray-induced rose mutants and assessment of genetic diversity

  • Chakrabarty, D.;Datta, S.K.
    • Plant Biotechnology Reports
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    • v.4 no.3
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    • pp.237-242
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    • 2010
  • Six parent and their 12 gamma ray-induced somatic flower colour mutants of garden rose were characterized to discriminate the mutants from their respective parents and understanding the genetic diversity using Random amplification of polymorphic DNA (RAPD) markers. Out of 20 primers screened, 14 primers yielded completely identical fragments patterns. The other 7 primers gave highly polymorphic banding patterns among the radiomutants. All the cultivars were identified by using only 7 primers. Moreover, individual mutants were also distinguished by unique RAPD marker bands. Based on the presence or absence of the 48 polymorphic bands, the genetic variations within and among the 18 cultivars were measured. Genetic distance between all 18 cultivars varied from 0.40 to 0.91, as revealed by Jaccard's coefficient matrix. A dendrogram was constructed based on the similarity matrix using the Neighbor Joining Tree method showed three main clusters. The present RAPD analysis can be used not only for estimating genetic diversity present in gamma ray-induced mutants but also for correct identification of mutant/new varieties for their legal protection under plant variety rights.

Rapid Detection and Isolation of Known and Putative $\alpha-L-Arabinofuranosidase$ Genes Using Degenerate PCR Primers

  • Park, Jung-Mi;Han, Nam-Soo;Kim, Tae-Jip
    • Journal of Microbiology and Biotechnology
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    • v.17 no.3
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    • pp.481-489
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    • 2007
  • [ $\alpha$ ]-L-Arabinofuranosidases (AFases; EC 3.2.1.55) are exo-type enzymes, which hydrolyze terminal nonreducing arabinose residues from various polysaccharides such as arabinan and arabinoxylan. Genome-wide BLAST search showed that various bacterial strains possess the putative AFase genes with well-conserved motif sequences at the nucleotide and amino acid sequence levels. In this study, two sets of degenerate PCR primers were designed and tested to detect putative AFase genes, based on their three highly conserved amino acid blocks (PGGNFV, GNEMDG; and DEWNVW). Among 20 Bacillus-associated species, 13 species were revealed to have putative AFase genes in their genome and they share over 67% of amino acid identities with each other. Based on the partial sequence obtained from an isolate, an AFase from Geobacillus sp. was cloned and expressed in E. coli. Enzymatic characterization has verified that the resulting enzyme corresponds to a typical AFase. Accordingly, degenerate PCR primers developed in this work can be used for fast, easy, and specific detection and isolation of putative AFase genes from bacterial cells.

Analysis of Genetic Relationships in Hypericum erectum Thunb. by RAPD (RAPD 방법을 이용한 고추나물의 유연관계분석)

  • Kim, Sun-Hee;Kim, Eung-Sik;Kim, Sung-Ho;Ahn, Jun-Cheul;Hwang, Baik
    • Korean Journal of Medicinal Crop Science
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    • v.13 no.4
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    • pp.141-145
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    • 2005
  • The genetic relationships of Hypericum erectum, H. ascyron and H. perforatum Thunb. by RAPD using total 46 primers were analysed 30 primers among primers tested showed the amplification band in all. The amplified DNA fragment ranged from 0.25 to 6.6 kb in size. The 411 bands (34.4%) among 1194 bands derived from 30 primers were polymorphic, and 13.7 bands per primer were observed. The phenograms for six analyzed individuals by RAPD markers were not matched well with those of the result by morphological characters. They were clustered monophyletic at the similarity coefficient value ranged from 0.24 to 0.96.

DNA Fingerprinting of Jute Germplasm by RAPD

  • Hossain, Mohammad Belayat;Haque, Samiul;Khan, Haseena
    • BMB Reports
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    • v.35 no.4
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    • pp.414-419
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    • 2002
  • The genotype characteristic of cultivars was investigated, along with varieties of both of the jute species, Corchorus olitorius and Corchorus capsularis, in the germplasm collection at the Bangladesh Jute Research Institute (BJRI). DNA fingerprinting was generated for 9 different varieties and 12 accessions of jute cultivars by using random amplified polymorphic DNA(RAPD). A total of 29 arbitrary oligonucleotide primers were screened. Seven primers gave polymorphism within the varieties, and 6 primers detected polymorphism within the accessions that were tested. A dendrogram was engendered from these data, and this gave a distinct clustering of the cultivated species of jute. Therefore, we generated RAPD markers, which are species-specific. These primers can distinguish between C. olitorius and C. capsularis. From the dendrogram that we generated between the various members of these two species, we found the existing genetic classification that agrees with our molecular marking data. A different dendrogram showed that jute accessions could be clustered into three groups. These data will be invaluable in the conservation and utilization of the genetic pool in the germplasm collection.

Rapid Screening of Apple mosaic virus in Cultivated Apples by RT-PCR

  • Ryu, Ki-Hyun;Park, Sun-Hee
    • The Plant Pathology Journal
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    • v.19 no.3
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    • pp.159-161
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    • 2003
  • The coat protein (CP) gene of Apple mosaic virus (ApMV), a member of the genus Ilarvirus, was selected for the design of virus-specific primers for amplification and molecular detection of the virus in cultivated apple. A combined assay of reverse transcription and polymerase chain reaction (RT-PCR) was performed with a single pair of ApMV-specific primers and crude nucleic acid extracts from virus-infected apple for rapid detection of the virus. The PCR product was verified by restriction mapping analysis and by sequence determination. The lowest concentration of template viral RNA required for detection was 100 fg. This indicates that the RT-PCR for detection of the virus is a 10$^3$times more sensitive, reproducible and time-saving method than the enzyme-linked immunosorbent assay. The specificity of the primers was verified using other unrelated viral RNAs. No PCR product was observed when Cucumber mosaic virus (Cucumovirus) or a crude extract of healthy apple was used as a template in RT-PCR with the same primers. The PCR product (669 bp) of the CP gene of the virus was cloned into the plasmid vector and result-ant recombinant (pAPCP1) was selected for molecule of apple transformation to breed virus-resistant transgenic apple plants as the next step. This method can be useful for early stage screening of in vitro plantlet and genetic resources of resistant cultivar of apple plants.

19th Century Chemistry Book of Korean Mathematician Sang-Seol LEE (한국 근대수학교육의 아버지 이상설(李相卨)이 쓴 19세기 근대화학 강의록 『화학계몽초(化學啓蒙抄)』)

  • Son, Yongkeun;Kim, Chae Sik;Lee, Sang-Gu;Lee, Jae Hwa
    • Korean Journal of Mathematics
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    • v.20 no.4
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    • pp.541-563
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    • 2012
  • Sang-Seol LEE wrote a manuscript HwaHakGyeMongCho(化學啓蒙抄) in the late 19th century. HwaHakGyeMongCho was transcribed from Science Primers: Chemistry (written by H. E. Roscoe), which is translated into Chinese by Joseph Edkins in 1886. LEE did not copy original writing exactly, but he understood the contents of each chapter and sections, then summarized and edited them in his caligraphic writing. In this paper, we introduce the contents for the first time and discuss the significance of this book.

Reverse Transcription Polymerase Chain Reaction-based System for Simultaneous Detection of Multiple Lily-infecting Viruses

  • Kwon, Ji Yeon;Ryu, Ki Hyun;Choi, Sun Hee
    • The Plant Pathology Journal
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    • v.29 no.3
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    • pp.338-343
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    • 2013
  • A detection system based on a multiplex reverse transcription (RT) polymerase chain reaction (PCR) was developed to simultaneously identify multiple viruses in the lily plant. The most common viruses infecting lily plants are the cucumber mosaic virus (CMV), lily mottle virus (LMoV), lily symptomless virus (LSV). Leaf samples were collected at lily-cultivation facilities located in the Kangwon province of Korea and used to evaluate the detection system. Simplex and multiplex RT-PCR were performed using virus-specific primers to detect single- or mixed viral infections in lily plants. Our results demonstrate the selective detection of 3 different viruses (CMV, LMoV and LSV) by using specific primers as well as the potential of simultaneously detecting 2 or 3 different viruses in lily plants with mixed infections. Three sets of primers for each target virus, and one set of internal control primers were used to evaluate the detection system for efficiency, reliability, and reproducibility.

A Study on Genetic Relationship between Sasang Constitutions by the Polymorphic Analysis of RAPD (RAPD 다형성 분석을 통한 사상체질간 유전적 상관관계에 관한 연구)

  • 이휘철;조동욱;조중호;서영우;이창수
    • The Journal of Korean Medicine
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    • v.20 no.4
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    • pp.62-68
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    • 2000
  • This study was carried out to establish genetic understanding of three Sasang constitutions of Taeumin, Soeumin and Soyangin by Random Amplified Polymorphic DNA(RAPD) analysis. We have applied RAPD analysis to pooled DNA sample as a means to achieve rapid screening of large numbers of primers for their capacity to reveal constitutions-specific polymorphisms. From an initial 440 primers, 13 polymorphic primers between different constitutions were selected. Bandsharing(BS) and mean average percentage difference(MAPD) calculated within and between three constitutions using RAPD fingerprint data showed a higher degree of homogenity within than between the constitutions and indicated measurable divergence between three constitutions. The RAPD bandsharing(BS) values ranged from 0.71 to 0.73 between the three constitutions. The interconstitution divergence was narrower between Taeumin and Soeumin, than between the other paired constitution comparisons. The genetic distance between the three constitutions was measured by BS values. Genetic distance by RAPD analysis was 0.007 between Taeumin and Soeumin, and 0.014 between Soyang and the others. In conclusion, the genetic distance of Teaumin and Soumin was closer than that of Soyangin in the analysis of RAPD by using 440 primers.

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Development of Gene Based STS Markers in Wheat

  • Lee, Sang-Kyu;Heo, Hwa-Young;Kwon, Young-Up;Lee, Byung-Moo
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.57 no.1
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    • pp.71-77
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    • 2012
  • The objective of this study is to develop the gene based sequence tagged site (STS) markers in wheat. The euchromatin enriched genomic library was constructed and the STS primer sets were designed using gene based DNA sequence. The euchromatin enriched genomic (EEG) DNA library in wheat was constructed using the $Mcr$A and $Mcr$BC system in $DH5{\alpha}$ cell. The 2,166 EEG colonies have been constructed by methylated DNA exclusion. Among the colonies, 606 colonies with the size between 400 and 1200 bp of PCR products were selected for sequencing. In order to develop the gene based STS primers, blast analysis comparing between wheat genetic information and rice genome sequence was employed. The 227 STS primers mainly matched on $Triticum$ $aestivum$ (hexaploid), $Triticum$ $turgidum$ (tetraploid), $Aegilops$ (diploid), and other plants. The polymorphisms were detected in PCR products after digestion with restriction enzymes. The eight STS markers that showed 32 polymorphisms in twelve wheat genotypes were developed using 227 STS primers. The STS primers analysis will be useful for generation of informative molecular markers in wheat. Development of gene based STS marker is to identify the genetic function through cloning of target gene and find the new allele of target trait.