• Title/Summary/Keyword: Transposon

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Establishment of An Efficient and Stable Transgene Expression System in Chicken Primordial Germ Cells

  • Yang, Ju-Hyun;Kim, Sung-Tae
    • Bulletin of the Korean Chemical Society
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    • v.33 no.5
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    • pp.1536-1540
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    • 2012
  • Chicken primordial germ cells (cPGCs) are founder germ cells in embryonic stage of development that eventually give rise to sperms or oocytes. Currently cPGCs are only known cells enabling germline transmission in chicken and their cultivation protocols were recently established. Although genome modifications of chickens are now theoretically possible using cPGCs, there are still several hurdles to overcome to practically use cPGCs as mediators for chicken transgenesis. First, efficiency of gene delivery into cPGCs remains low with current methods. Second, there aregene silencing mechanisms against the expression of foreign genes in cPGCs. In this study, we successfully increased the efficiency of gene delivery in cPGCs by taking advantage of the TTAA-specific $piggybac$ transposon system. Moreover, a pipette-type electroporator significantly enhanced transfection efficiency up to 5-fold compared withcuvette-type methods. Taken together, the technological advances in our study will provide practical benefits for the application to fulfill genetic modifications of chicken genome.

Genetic Organization of the Recombinant Bacillus pasteurii Urease Genes Expressed in Escherichia coli

  • Kim, Sang-Dal;Hausinger, Robert P.
    • Journal of Microbiology and Biotechnology
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    • v.4 no.2
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    • pp.108-112
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    • 1994
  • The genetic organization of the urease gene cluster from an alkalophilic Bacillus pasteurii was determined by subcloning and Tn5 transposon mutagenesis of a 10.7 kilobasepair cloned fragment. A region of DNA between 5.0 and 6.0 kb in length is necessary for urease activity. In vitro transcription-translation analysis of transposon insertion mutants of the cloned urease genes demonstrated that the major ($M_r$ 67,000) and minor ($M_r$ 20,000) structural peptides of urease are encoded at one end of the urease gene cluster and at least 3 additional polypeptides are encoded by adjacent DNA sequences.

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Cloning of agrobacterium tumefaciens chromosomal virulence region (Agrobacterium tumefaciens의 염색체 DNA내에 존재하는 종양 유발 지역의 클로닝)

  • ;Cangelosi, G.A.;Nester, E.W.
    • Korean Journal of Microbiology
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    • v.28 no.2
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    • pp.104-108
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    • 1990
  • The chromosomal DNA of Agrobacterium tumefaciens contains the genes required for bacterial attachment to plant cell which is an essential atage in crown gall tumorigenesis by Ti-plasmid. In order to clone the genes, Agrobacterium tumefaciens strain A5512 was mutagenized by transposon Tn5 and two Agrobacterium tumefaciens mutants which are attachment-defective and nontumorigenic were isolated. From one of the two mutants, a chromosomal virulence region which was required for attachment to the plant cells was cloned.

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Integration and Expression of BaciZlus thun'ngiensis Crystal Protein Gene in Chromosomal DNA of Pseudomonas Strains Using Transposon Tn5 (Transposon Tn5에 의한 Bacillus thuringiensis 독소단백질 유전자의 Pseudomonas 내로의 도입 및 발현)

  • 신병식;구본탁;박승환;김정일
    • Microbiology and Biotechnology Letters
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    • v.19 no.1
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    • pp.25-30
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    • 1991
  • The crystal protein gene (cp) of Bacillus tizuringienszs subsp. liuvstaki (B.t.k.) HI173 was subcloned into HanzHI site of central region (Tn5-cp) or BglII site of IS50L region (IS50L-cp) in Tn5, and transposed into the chromosomal DNA of five strains of root-colonizing Pseudomonas. The expression of cp gene in Acwiomoncrs transconjugants was demonstrated by immunoblot analysis and bioassay against larvae of the Hyphantria cunea.

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Zoolan Gene Cloning of Zoogloea ramigera 115 (Zoogloea ramigera 115의 Zooglan Gene Cloning)

  • 이기영;전순배
    • KSBB Journal
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    • v.11 no.1
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    • pp.115-123
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    • 1996
  • Two kinds of mutants were isolated to clone a cluster of genes essential for zooglan biosynthesis. Zoogloea ramigera 115 strains produce capsular polysaccharide. To achieve conjugation in strain 115 and to facilitate recovery of product, a capsule non-forming strain was isolated via successive centrifugation and screening. The other kind of mutants devoid of or producing altered exopolysaccharides were obtained using classical transposon(Tn5) technique and screened for altered colony morphology and celluflour binding properties. Complementation of these mutants was achieved with Z. ramigera 115 slime gene library constructed in a broad host range cosmid vector and helper plasmid by triparental conjugation.

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