• 제목/요약/키워드: marker selection

검색결과 510건 처리시간 0.028초

Development of Gene-based Markers for the Allelic Selection of the Restorer-of-fertility Gene, Rfo, in Radish (Raphanus sativus)

  • Kim, Sunggil;Lim, Heerae;Cho, Kang-Hee;Park, Pue Hee;Park, Suhyung;Sung, Soon-Kee;Oh, Daegeun;Kim, Ki-Taek
    • Korean Journal of Breeding Science
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    • 제41권3호
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    • pp.194-204
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    • 2009
  • Cytoplasmic male sterility (CMS) and fertility restoration have been utilized as valuable tools for $F_1$-hybrid seed production in many crops despite laborious breeding processes. Molecular markers for the selection of CMS-related genes help reduce the expenses and breeding times. A previously reported genomic region containing the Ppr-B gene, which is responsible for restoration of fertility and corresponds to the Rfo locus, was used to develop gene-based or so-called "functional" markers for allelic selection of the restorer-of-fertility gene (Rfo) in $F_1$-hybrid breeding of radish (Raphanus sativus L.) Polymorphic sequences among Rfo alleles of diverse breeding lines of radish were examined by sequencing the Ppr-B alleles. However, presence of Ppr-B homolog, designated as Ppr-D, interferes on specific PCR amplification of Ppr-B in certain breeding lines. The organization of Ppr-D, resolved by genome walking, revealed extended homology with Ppr-B even in the promoter region. Interestingly, PCR amplification of Ppr-D was repeatedly unsuccessful in certain breeding lines implying the lack of Ppr-D in these radishes. Ppr-B could only be successfully amplified for analysis through designing primers based on the sequences unique to Ppr-B that exclude interference from Ppr-D gene. Four variants of Rfo alleles were identified from 20 breeding lines. A combination of three molecular markers was developed in order to genotype the Rfo locus based on polymorphisms among four different variants. These markers will be useful in facilitating $F_1$-hybrid cultivar development in radish.

Assessment of Factors Influencing Agrobacterium Mediated Transformation in Cucumber (Cucumis sativus L)

  • Sureshkumar P.;Selvaraj N.;Ganapathi A.;Kasthurirengan S.;Vasudevan A.;Anbazhagan V. Ramesh
    • Journal of Plant Biotechnology
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    • 제7권4호
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    • pp.225-231
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    • 2005
  • Five day old cotyledon explants of Cucumber (Cucumis sativus L) cv Poinsett 76 were cocultivated with two Agrobacterium strains (EHA105 and LBA 4404) each carrying GUS as the reporter gene and npt-II as the selection marker gene in the T-DNA region of the vector. Transformed shoots were selected at 150 mg/L kanamycin. A two day cocultivation coupled with $20\;{\mu}M$ acetosyringone increased the frequency (8.2 and 15.4 shoots) of GUS expression in the shoots of transformed plant. Among the two Agrobacterium strains, EHA 105 performed better than LBA 4404 in bringing two-fold increase in transformation efficiency (14%) than LBA 4404 (7.4%). PCR analysis was done to confirm the integration of T-DNA into cucumber genome.

Risk Selection and Tumor Marker (위험선택과 종양표식자 - CEA와 AFP에 대해서 -)

  • Park, Sung-Soo;Lee, Shin-Whi
    • The Journal of the Korean life insurance medical association
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    • 제17권
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    • pp.77-83
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    • 1998
  • 당사에서는 악성 종양에 대한 위험선택의 수단으로써 1997년 1월부터 종양표식자 중 CEA와 AFP 검사를 사의 진단에 한하여 실시하고 있다. 검사의 실시조건은 의사의 진찰소견으로 필요하다고 인정되는 경우와 관련병력의 고지가 있는 경우로 한하였다. 1997년 1월부터 10월말까지 실시한 건에 대해서만 분석을 하였으며 CEA 534건, AFP 608건를 실시하였다. 그 결과 기준치를 초과하는 경우는 CEA 33건, AFP 23건이었다. 표준체로 판정된 경우의 CEA의 평균치는 1.5ng/ml, AFP는 2.6ng/ml였다. CEA와 AFP의 검사결과나 그 이외의 원인으로 사절체의 판정을 받은 경우의 CEA의 평균치는 3.2ng/ml AFP의 경우는 20.7ng/ml로 나타났다. 본연구에서는 생명보험회사에서 종양표식자 검사의 의미를 검토해 보고자 하였다.

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Agrobacterium-mediated transformation of Lycopersicon esculentum (cv. MicroTom) with two pathogen-induced hot pepper transcription factors

  • Seong, Eun-Soo;Oh, Sang-Keun;Eunsook Chung;Park, Doil
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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    • pp.79.1-79
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    • 2003
  • Two pathogen-induced hot pepper transcription factors (CaNACl and CapIfl) were introduced into‘MicroTom’tomato by Agrobacterium tumefaciens-mediated transformation. We used to nptII containing kanamycin resistance gene as a selection marker. Both transformed and non-transformed plants were transferred to pot after rooting test in vitro. To approximate the levels of caNACl transcript in leaves of wild-type and transgenic plants, RNA blots were hybridized with double-stranded full-length CaNACl probe at moderate stringency, Although the relative signal strength for hybridization fluctuated among the samples on different blots, transgenic plant lines N-1, N-2 and N-3 consistently displayed increased levels of CaNACl transcript relative to other transgenic lines and wild-type plants. Of all the transgenic lines examined, line N-7 had the least amount of CaNACl transcript. Role of these transcription factors in pathogen defense will be examined by overexpression in tomato.

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Effect of Sonication and vir Genes on Transient Gene Expression in Agrobacterium-Mediated Transformation (Agrobacterium을 이용한 형질전환에서 sonication과 vir 유전자들의 효과)

  • 이병무
    • Journal of Life Science
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    • 제11권4호
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    • pp.316-320
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    • 2001
  • Sonication tremendously improves the efficiency of Agrobacterium infection by introducing small and uniform fissures and channels throughout the targeted tissue. Using shoot tips of cotton as explants, the effect of sonication treatment and virulence genes in Agrobacterium tumefaciens on transformation efficiency was investigated. The pat gene which encodes resistance to the herbicide, glufosinate, was used as a selectable marker. Transformation efficiency was evaluated on th basis of survival rates of cocultivated shoot tips on selection medium containing 2.5 mg/l gulfosinate-ammonium(ppt) adn 25. mg/l Clavamax. Sonication from 5 to 15 second has a positive effect on shoop tip survival. However, whil virE as well as virG or vir GN54D showed an enhancement in transformation efficiency, virE,. virG resulted in the most significant enhancement. Overall, the combination of additional virG/virE gene and sonication treatment resulted in the most significant increase in transformation efficiency.

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Genomic and Transgenic Approaches to Modified Plants: Disease Resistance in the Brassica as a Model System.

  • Ekuere, Usukuma;Good, Allen G.;Mayerhofer, Reinhold
    • Korean Journal of Plant Tissue Culture
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    • 제27권4호
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    • pp.317-323
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    • 2000
  • Molecular genetic techniques can now be applied to the development of advanced plant genotypes, either through genetic transformation or genomic approaches which allow researchers to transfer specific traits using molecular markers. In this paper, we discuss the use of these techniques towards understanding the genetics of blackleg resistance in Brassica. In a comparative mapping study between Arabidopsis thaliana and Brassica napus, 6 R-ESTs, 7 B. napus RFLP markers and a B. napus EST were located in a collinear region of N7 (B. napus) and chromosome 1 (A. thaliana). One of the A. thaliana R-ESTs and 4 of the B. napus RFLPs co-segregated and mapped to the LmRl locus for blackleg resistance. Introgression of blackleg resistance from wild relatives is also investigated with the possibility of accelerating the introgression process via marker assisted selection.

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Agrobacterium-Mediated Co-transformation of Multiple Genes in Metarhizium robertsii

  • Padilla-Guerrero, Israel Enrique;Bidochka, Michael J.
    • Mycobiology
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    • 제45권2호
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    • pp.84-89
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    • 2017
  • Fungi of the Metarhizium genus are a very versatile model for understanding pathogenicity in insects and their symbiotic relationship with plants. To establish a co-transformation system for the transformation of multiple M. robertsii genes using Agrobacterium tumefaciens, we evaluated whether the antibiotic nourseothricin has the same marker selection efficiency as phosphinothricin using separate vectors. Subsequently, in the two vectors containing the nourseothricin and phosphinothricin resistance cassettes were inserted eGFP and mCherry expression cassettes, respectively. These new vectors were then introduced independently into A. tumefaciens and used to transform M. robertsii either in independent events or in one single co-transformation event using an equimolar mixture of A. tumefaciens cultures. The number of transformants obtained by co-transformation was similar to that obtained by the individual transformation events. This method provides an additional strategy for the simultaneous insertion of multiple genes into M. robertsii.

TRANSMISSION OF C-BAND VARIANTS IN JAPANESE QUAIL

  • Sohn, S.H.;Fechheimer, N.S.;Nestor, K.E.
    • Asian-Australasian Journal of Animal Sciences
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    • 제8권2호
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    • pp.171-174
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    • 1995
  • Heteromorphisms of chromosome banding patterns can be useful markers for gene mapping and other kinds of genetic studies. In Japanese quail, the centromere region of chromosome No. 4 is the site of a heteromorphism. One form of the C-band at this region is relatively small ("a" form); an alternative form is much larger ("b" form). To identify the transmission patterns, all possible matings were made between birds with karyotype a/a, a/b, and b/b. The outcome from all crosses are entirely consistent with the expectation from simple Mendelian transmission. No evidence was found for segregation distortion or gametic selection. This dimorphism, therefore, is a reliable marker.

Association of a Single Nucleotide Polymorphism in the 5'-Flanking Region of Porcine HSP70.2 with Backfat Thickness in Duroc Breed

  • Chen, Ming-Yu;Huang, San-Yuan;Lin, En-Chung;Hseu, Tzong-Hsiung;Lee, Wen-Chuan
    • Asian-Australasian Journal of Animal Sciences
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    • 제16권1호
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    • pp.100-103
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    • 2003
  • Higher environmental temperature affects the economic performance of pigs. Heat shock protein 70 has been shown to play an important role in thermoresistance. The purpose of this study was to assess the effect of a single nucleotide polymorphism in the 5'-flanking region of porcine HSP70.2 on growth performance in Taiwanese Duroc. The genotype of this nt 393 polymorphic site could be verified by digestion with Bsa WI restriction enzyme of a PCR product. Pigs with TT and TC genotypes have thinner backfats than those with CC type (p<0.05). The result suggested that the polymorphic Bsa WI site in the 5'flanking region of porcine HSP70.2 may be used as a marker for the early selection of ultrasonic backfat thickness in Duroc pigs.

Lactobacillus casei YIT 9018의 Shuttle Vector 개발을 위한 분자유전학적 연구

  • Yoo, Min;Nam, Jin-Sik;Kwon, Oh-Sik;Baek, Young-Jin
    • Microbiology and Biotechnology Letters
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    • 제25권5호
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    • pp.464-467
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    • 1997
  • A shuttle vector, pSHvec, was constructed for Lactobacillus casei (L. casei) YIT 9018 and JM1O9 by recombinant DNA technology. This vector contained the $\beta$-lactamase II gene from Bacillus cereus as a selection marker and replication origins for both Gram(+) and Gram(-) strains. It could transform the wild type L. casei YIT 9018 as well as E. coli JM109 and transformed cells were selected based on antibiotics resistance. The ability of L. casei YIT 9018 for curd formation in 11% skim milk was maintained even after transformation with pSHvec. The vector was stable as long as antibiotics were added to the medium. These results could contribute to the study of lactic acid bacteria for the industrial purpose at a genetic level.

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