• Title/Summary/Keyword: Germ line transformation

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Transposon piggyBac mediated Ipax6 Expression in Malaria Vector Anopheles stephensi (말라리아 매개 모기 Anopheles stephensi에서 트랜스포존 piggyBac을 이용한 Pax6 발현)

  • Koo Hyeyoung
    • Development and Reproduction
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    • v.8 no.1
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    • pp.19-25
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    • 2004
  • Pax6, a member of the highly conserved homeobox gene family, is known to be expressed in spatially and temporally restricted pattern during embryogenesis. To examine the spatial expression pattern of Pax6 in malaria vector mosquito Anopheles stephemi, in different molecular environment, the germ line transformation technique using piggyBac transposon combined with the use of Pax6 specific 3xp3-EGFP marker was utilized. Four transgenic lines with a transformation rate of 6.7% were established. Transgenes were stably expressed in subsequent several generations. The transgenic lines showed 3 different expression pattern with spatial specificity, possibly due to enhancing and/or silencing position effects. In two transgenic lines, noble expression pattern of Pax6 was observed in the region that has not been previously reported in any animal species. The results show that the tranposon piggyBac mediated germ line transformation system can be used as an efficient tool for the generation of diverse spatially restricted reporter gene expression.

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Germ Line Transformation of the Silkworm, Bombyx mori L. with a piggyBac Vector Harboring the Human Lactoferrin Gene (락토페린 유전자도입 piggyBac 벡터에 의한 누에 형질전환)

  • Kim, Yong-Soon;Sohn, Bong-Hee;Kim, Kee-Young;Jung, I-Yeon;Kim, Mi-Ja;Kang, Pil-Don
    • Journal of Sericultural and Entomological Science
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    • v.49 no.2
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    • pp.37-42
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    • 2007
  • Lactoferrin, an ion-binding 80-kDa glycoprotein, has been suggested to have many biologic activities, such as facilitating ion absorption and having antimicrobial and anti-inflammatory effects. Several of these activities are likely to only be facilitated by human lactoferrin because they depend on the binding of human lactoferrin to specific receptor. To produce recombinant human lactoferrin to animal foods using transgenic silkworm, Bombyx mori L, we have cloned and sequenced the cDNA encoding for a human lactoferrin (HLf) from the mRNA in mammary tumor line (GI-101). As a result, the 2.5-kb fragment of HLf gene was cloned with pGEM-T vector and then this fragment was sequenced. In the nucleotide sequence analysis, single open reading frame of the 2,136-bp encoding for a polypeptide of 712 amino acid residues was detected. On the other hand, we constructed a recombinant plasmid(pPT-HLf), containing human lactoferrin gene for germ line transformation of the silkworm using a piggyBac transposon-derived vector. A nonautonomous helper plasmid encodes the piggyBac transposase. Approximately 6.7% of individuals in the G0 silkworms expressed green fluorescent protein (GFP). PCR analyses of GFP-positive silkworms (G0 and G1) revealed that independent insertions occurred frequently. Furthermore, Western blot analysis showed that the recombinant HLf expressed in hemolymph has the same molecular weight (80 kDa) as a native protein. On the basis of these experiments, expression of HLf in next generation of transgenic silkworm is now in process.

Probing Cell-Type Specific Gene Expression in the Ovarian Cells of Drosophila by P-Element Mediated Enhancer Detection (P-요소를 이용한 노랑초파리 난소에서의 세포특이적 유전자발현의 검출)

  • 계명찬;조경상;김경진;이정주
    • The Korean Journal of Zoology
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    • v.38 no.4
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    • pp.505-513
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    • 1995
  • P-lement mediated enhancer detector lines (EDla) were screened for reporter gene (1acZ expression In the ovary of Drosophila mejanogaster Cell-type spedfic 1acZ expression can be grouped Into three parts such as in the geimline, soma, and both. LacZ expression In germline cells was devided into 2 types; expression in nurse cells or in both of the nurse cells and oocote. In the stage-9 to stage-lO follicles, lacZ expression was observed either In the whole follicle cells around oocote or in the subpopulation of follicle cells in egg chamber. lacZ expression in the subset of follicle cells are showed in the centripetal follicle cells or the columnar follicle cells except centripetal follicle cells. Several lines showed anterior to postedor gradient pattern of lacZ expression in the follicle cells. Interestingly there were 3 lines in which lacZ was expressed In the polar cells and/or the horder cells of egg chamber. These lacZ expression patterns in the different ovarian cells of independent EDla reflect the cell type-spedflc expression of maternal genes nesr the P-element insertion, and might provide a basis for cloning of genes involved in oogenesis of Drosophila.

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