• 제목/요약/키워드: genomic integration

검색결과 101건 처리시간 0.027초

Insertional mutagenesis of fusarium graminearum for characterization of genes involved in disease development and mycotoxin production

  • Han, Yon-Kyoung;Lee, Hyo-Jin;Yun, Sung-Hwan;Lee, Yin-Won
    • 한국식물병리학회:학술대회논문집
    • /
    • 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
    • /
    • pp.85.2-86
    • /
    • 2003
  • Fusarium graminearum is an important pathogen of cereal crops in many areas of the world causing head blight and ear rot of small grains. In addition to serious economic losses, this fungus produces mycotoxins, such as trichothecenes and zearalenone on diseased crops and has been a potential threat to human and animal health. To massively identify pathogenesis-related genes from F. graminearum, two representative strains (SCKO4 from rice and Z03643 from wheat) were mutagenized using restriction enzyme-mediated integration (REMI). In total, 20,DOD REMI transformants have been collected from the two strains. So far, 63 mutants for several traits involved in disease development such as virulence, mycotoxin production, and sporulation have been selected from 3,000 REMI transformants. Now, selected mutants of interest have being genetically analyzed using a newly developed outcross method (See Jungkwan Lee et al poster). In addition, cloning and characterization of genomic DNA regions flanking the insertional site in the genome of the mutants are in progress.

  • PDF

Generation of an Arginine Auxotrophic Mutant of Colletotrichum acutatum as a Recipient Host for Insertional Mutagenesis

  • Kim, Hee-Kyoung;Lee, Sun-Hee;Kim, Heung-Tae;Yun, Sung-Hwan
    • The Plant Pathology Journal
    • /
    • 제25권3호
    • /
    • pp.205-212
    • /
    • 2009
  • Colletotrichum acutatum was the main cause of the recent outbreaks of anthracnose on pepper fruit in Korea. To facilitate molecular analysis of C. acutatum, we generated an arginine auxotrophic mutant of the C acutatum strain JC24 using a targeted gene replacement strategy. A 3.3-kb genomic region carrying an ortholog (designated CaARG2) of the fungal gene encoding N-acetylglutamate synthase, the first enzyme of arginine biosynthesis in fungi, was deleted from the fungal genome. The mutant exhibited normal growth only when arginine was exogenously supplied into the culture medium. Transformation of the arginine auxotrophic mutant with a plasmid DNA carrying an intact copy of CaARG2, which was smaller than the deleted region in the mutant, not only caused random vector insertions in the fungal genome, but also recovered both hyphal growth and pathogenicity of the mutant to the wild-type level. Using this new selection system, we have successfully developed a restriction enzyme-mediated integration procedure, which would provide an economically efficient random mutagenesis method in C. acutatum.

From the Sequence to Cell Modeling: Comprehensive Functional Genomics in Escherichia coli

  • Mori, Hirotada
    • BMB Reports
    • /
    • 제37권1호
    • /
    • pp.83-92
    • /
    • 2004
  • As a result of the enormous amount of information that has been collected with E. coli over the past half century (e.g. genome sequence, mutant phenotypes, metabolic and regulatory networks, etc.), we now have detailed knowledge about gene regulation, protein activity, several hundred enzyme reactions, metabolic pathways, macromolecular machines, and regulatory interactions for this model organism. However, understanding how all these processes interact to form a living cell will require further characterization, quantification, data integration, and mathematical modeling, systems biology. No organism can rival E. coli with respect to the amount of available basic information and experimental tractability for the technologies needed for this undertaking. A focused, systematic effort to understand the E. coli cell will accelerate the development of new post-genomic technologies, including both experimental and computational tools. It will also lead to new technologies that will be applicable to other organisms, from microbes to plants, animals, and humans. E. coli is not only the best studied free-living model organism, but is also an extensively used microbe for industrial applications, especially for the production of small molecules of interest. It is an excellent representative of Gram-negative commensal bacteria. E. coli may represent a perfect model organism for systems biology that is aimed at elucidating both its free-living and commensal life-styles, which should open the door to whole-cell modeling and simulation.

Increased Thermotolerance of Transgenic Rice Plant by Introduction of Thermotolerant Gene

  • Lee, Byung-Hyun;Won, Sung-Hye;Kim, Ki-Yong;Lee, Hyoshin;Jinki Jo
    • 한국초지조사료학회지
    • /
    • 제20권2호
    • /
    • pp.97-102
    • /
    • 2000
  • To increase thennotolerance of forage crops, transgenic rice plants as a model for transformation of monocots were generated. A cDNA encoding the chloroplast-localized small heat shock protein (small HSP) of rice, Oshsp21, was introduced into rice plants via Agrobacterium-mediated gene transfer system. Calli induced from scutella were co-cultivated with a A. tumefaciens strain EHAlOl canying a plasmid, pIGhsp21. A large number of transgenic plants were regenerated on a medium containing hygromycin. Integration of Oshsp2l gene was confirmed by PCR and Southern blot analyses with genomic DNA. Northern blot and immunoblot analyses revealed that the Oshsp21 gene was constitutively expressed and accumulated as mature protein in transgenic plants. Effects of constitutive expression of the OshspZl on thermotolerance were first probed with the chlorophyll fluorescence. Results indicate that inactivation of electron transport reactions in photosystem I1 (PSII), were mitigated by constitutive expression of the Oshsp21. These results suggest that the chloroplast small HSP plays an important role in protecting photosynthetic machinery during heat stress. (Key words : Thermotolerance, Rice, Transgenic, cDNA)

  • PDF

Mutational Data Loading Routines for Human Genome Databases: the BRCA1 Case

  • Van Der Kroon, Matthijs;Ramirez, Ignacio Lereu;Levin, Ana M.;Pastor, Oscar;Brinkkemper, Sjaak
    • Journal of Computing Science and Engineering
    • /
    • 제4권4호
    • /
    • pp.291-312
    • /
    • 2010
  • The last decades a large amount of research has been done in the genomics domain which has and is generating terabytes, if not exabytes, of information stored globally in a very fragmented way. Different databases use different ways of storing the same data, resulting in undesired redundancy and restrained information transfer. Adding to this, keeping the existing databases consistent and data integrity maintained is mainly left to human intervention which in turn is very costly, both in time and money as well as error prone. Identifying a fixed conceptual dictionary in the form of a conceptual model thus seems crucial. This paper presents an effort to integrate the mutational data from the established genomic data source HGMD into a conceptual model driven database HGDB, thereby providing useful lessons to improve the already existing conceptual model of the human genome.

Isolation and Characterization of the Eicosapentaenoic Acid Biosynthesis Gene Cluster from Shewanella sp. BR-2

  • Lee, Su-Jin;Seo, Pil-Soo;Kim, Chul-Ho;Kwon, Oh-Suk;Hur, Byung-Ki;Seo, Jeong-Woo
    • Journal of Microbiology and Biotechnology
    • /
    • 제19권9호
    • /
    • pp.881-887
    • /
    • 2009
  • Forty-four eicosapentaenoic acid (EPA)-producing microbial strains were isolated from the intestines of marine fishes. Among them, one strain showing a maximum level of EPA (4.78% of total fatty acids) was identified as Shewanella sp. BR-2 on the basis of its 168 rRNA sequence. The EPA content reached a maximum level during the mid-exponential phase of cell growth, and gradually decreased with further growth of the cells. A cosmid DNA including the EPA biosynthesis gene cluster consisting of pfaA-E was isolated from a cosmid library of genomic DNA of Shewanella sp. BR-2, named pCosEPA-BR2. An E. coli clone harboring pCosEPA-BR2 produced EPA at a maximum level of 7.5% of total fatty acids, confirming the EPA biosynthesis activity of the cloned gene cluster.

A DNA Microarray LIMS System for Integral Genomic Analysis of Multi-Platform Microarrays

  • Cho, Mi-Kyung;Kang, Jason Jong-ho;Park, Hyun-Seok
    • Genomics & Informatics
    • /
    • 제5권2호
    • /
    • pp.83-87
    • /
    • 2007
  • The analysis of DNA microarray data is a rapidly evolving area of bioinformatics, and various types of microarray are emerging as some of the most exciting technologies for use in biological and clinical research. In recent years, microarray technology has been utilized in various applications such as the profiling of mRNAs, assessment of DNA copy number, genotyping, and detection of methylated sequences. However, the analysis of these heterogeneous microarray platform experiments does not need to be performed separately. Rather, these platforms can be co-analyzed in combination, for cross-validation. There are a number of separate laboratory information management systems (LIMS) that individually address some of the needs for each platform. However, to our knowledge there are no unified LIMS systems capable of organizing all of the information regarding multi-platform microarray experiments, while additionally integrating this information with tools to perform the analysis. In order to address these requirements, we developed a web-based LIMS system that provides an integrated framework for storing and analyzing microarray information generated by the various platforms. This system enables an easy integration of modules that transform, analyze and/or visualize multi-platform microarray data.

Antisense expression of a staygreen gene (SGR) delays leaf senescence in creeping bentgrass

  • Hwang, Ok-Jin;Han, Yun-Jeong;Paek, Nam-Chon;Kim, Jeong-Il
    • Rapid Communication in Photoscience
    • /
    • 제3권2호
    • /
    • pp.28-31
    • /
    • 2014
  • Loss of chlorophyll is the visible symptom of leaf senescence and staygreen refers to the delayed leaf senescence in plants. The staygreen gene (SGR) in rice (Oryza sativa L.) has been identified as its mutation maintains greenness during leaf senescence, and encodes a chloroplast protein required for the initiation of chlorophyll breakdown in plants. In this study, we isolated a rice SGR-homologous gene in creeping bentgrass (Agrostis stolonifera L.), and transgenic creeping bentgrass plants were obtained by introducing pCAMBIA3301 vector harboring antisense SGR gene under control of the senescence-specific SAG12 promoter. Transgenic plants were selected by herbicide resistance assays and genomic integration of the transgenes was confirmed by PCR analysis. Subsequent analyses demonstrated the staygreen phenotype of the transgenic creeping bentgrass plants with decreased chlorophyll loss during leaf senescence. These results suggest that the antisense SGR expression in creeping bentgrass delays leaf senescence, which provides a way to develop genetically engineered turfgrass varieties with the commercially useful staygreen trait.

The integration of genomics approaches for lettuce (Lactuca sativa L.) improvements on the disease resistances and other agronomic qualities.

  • Kim, Tae-Sung;Kim, Jeong-Haw;Kim, Jung-Bun;Jang, Suk-Woo
    • 한국작물학회:학술대회논문집
    • /
    • 한국작물학회 2017년도 9th Asian Crop Science Association conference
    • /
    • pp.114-114
    • /
    • 2017
  • The aim of this research is to improve Korean lettuce varieties in terms of Fusarium wilt, bolting under hot weather and nutritional function applying genomics approaches. To find related gene/molecular markers, we selected 96 lettuce varieties which are popular in domestic fresh vegetable markets. To construct frame works of the genomic approaches, we exploited GBS(Genotyping by Sequencing) and found total 61,407 SNPs from lettuce whole genomes (MAF>0.02). We observed that Three SNPs array per 100kb of lettuce genome. Average LD decay is expected to expand up to 3.9M(million)bp. Thus, we concluded that about 104 SNPs exist within a LD, which is sufficient to use GWAS(Genome-wide Association Study) to explore the useful gene/molecular markers. In addition, we optimized mass screening method to evaluate disease resistance levels against Fusarium wilt and are testing the bolting sensitivity during summer growing season for those lettuce allele mining set.

  • PDF

Current Status of Wheat Transformation

  • Rakszegi, Marrian;Tamas, Cecilia;Szucs, Peter;Tamas, Laszlo;Bedo, Zoltan
    • Journal of Plant Biotechnology
    • /
    • 제3권2호
    • /
    • pp.67-81
    • /
    • 2001
  • Traditionally, genetic variability is generated by an extensive crossing program, which is complemented by strict selection to identify useful new recombinants. Plant biotechnology offers many opportunities for breeders to solve certain breeding problems at the molecular level. The tissue culture methodology and the genetic modification of economically important monocotyledons have undergone a revolution in the last decade. As the production of transgenic plants is a complex procedure, including the uptake of DNA molecules into the cells, the integration of foreign nucleotide sequences into the host genomic DNA and the expression of new genes in a controlled way, and as there are still many unsolved questions, further development is necessary. The methodology opens up the possibility of introducing novel genes that may induce resistance to diseases and abiotic stresses, allow the modification of dough quality and the dietetic quality of proteins, and increase the levels of micronutrients such as iron, zinc, and vitamins. In the present review, the authors would like to summarise the most important advances in wheat transformation.

  • PDF