• Title/Summary/Keyword: 형질전환초파리

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Expression of Expanded Polyglutamine Disease Proteins in Drosophila (Drosophila Polyglutamine Disease Models) (증가된 글루타민에 의해 초래되는 뇌신경질환의 초파리 모델에 대한 연구)

  • Shin, Sang Min;Paik, Kyung Hoon;Jin, Dong Kyu
    • Clinical and Experimental Pediatrics
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    • v.48 no.4
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    • pp.425-432
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    • 2005
  • Purpose : Polyglutamine diseases are a group of diseases caused by the expansion of a polyglutamine tract in the protein. The present study was performed to verify if polyglutamine disease transgenic Drosophila models show similar dysfunctions as are seen in human patients. Methods : Polyglutamine disease transgenic Drosophila were tested for their climbing ability. And using genetic methods, the effects of anti-apoptotic gene bcl-2 and chemical chaperones on neurodegeneration were observed. Also, spinocerebellar ataxia 2 (SCA2) transgenic Drosophila lines were generated for future studies. Results : Expanded forms of spinocerebellar ataxia 3 (SCA3) transgenic protein causes characteristic locomotor dysfunction when expressed in the nervous system of Drosophila but the anti-apoptotic gene bcl-2 shows no evidence of ameliorating the deleterious effect of the expanded protein. However, Glycerol, a chemical chaperone, seemed to reduce the toxicity, at least in the eyes of the transgenic flies. The level SCA2 expression is too weak in the transgenic SCA2 Drosophila for evaluation. Conclusion : SCA3 transgenic Drosophila show ataxic behavior as observed in human patients. Chemical chaperones such as glycerol may prove beneficial in this class of genetic disease, which has no current method of cure.

Analysis of the Role of STAT Binding Site in the Drosophila raf Promoter Region Using Transgenic Flies (형질전환 초파리를 이용한 Drosophila raf 유전자 발현조절영역에 존재하는 STAT결합부위의 역할에 관한 연구)

  • Park, Hyun Sook;Kim, Young Shin;Kwon, Eun Jeong;Yoo, Mi Ae
    • Journal of Life Science
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    • v.9 no.1
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    • pp.50-57
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    • 1999
  • STATs activated by various cytokine and growth factors trigger a quick response in the nucleus and induce changes in gene expression. We have found the sequences homologous to STAT binding site in the 5'-flanking region of the D-raf gene. In this study, we examined role of the STAT binding site in D-raf gene promoter activity in vivo by using transgenic flies. The reporter plasmid pDraf-STATmut-lacZ was constructed by fusing D-raf promoter fragment having the base-substituted STAT binding site with the lacZ gene in a P-element vector. Transgenic flies bearing the Draf-STATmut-lacZ fusion genes were established by P-element mediated transformation. The expression of lacZ in transgenic flies bearing Draf-STATmut-lacZ fusion genes carrying base substitution in STAT site throughout various developmental stages was extensively reduced in comparison with that in transgenic flies bearing wild type Draf-lacZ fusion gene. These results show that the STAT binding site plays an important role in regulation of the D-raf gene.

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Molecular Mechanism of ABC Transporter Mdr49A Associated with a Positive Cross-Resistance in Transgenic Drosophila (형질전환 초파리를 이용한 Mdr49A 유전자의 살충제 교차저항성 기능 구명)

  • Seong, Keon Mook;Pittendrigh, Barry R.
    • Korean journal of applied entomology
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    • v.59 no.4
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    • pp.341-348
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    • 2020
  • The ATP-binding cassette (ABC) transporter superfamily represents the largest transmembrane protein that transports a variety of substrates across extra- and intra-cellular membranes. In insects, the ABC transporter proteins play crucial roles in insecticide resistance. To date, no studies have investigated the involvement of ABC transporter gene for cross-resistance to insecticide chemistries. Here, we studied such possible mechanisms against six conventional insecticides using transgenic Drosophila melanogaster strains carrying Mdr49 transcript variant A. For the 91-R and 91-C strains of Drosophila melanogaster, although they have a common origin, 91-R has been intensely selected with DDT for over 60 years, while 91-C has received no insecticide selection. Our transgenic analyses showed that overexpression of 91-R-MDR49 transcript variant A along with three amino acid variations can yield a relatively low degree of cross-resistance to carbofuran (2.0~6.7-fold) and permethrin (2.5~10.5-fold) but did not show cross-resistance to abamectin, imidacloprid, methoxychlor, and prothiofos as compared to the Gal4-driver control strain without transgene expression. These results indicate that the overexpression of Mdr49A in itself leads to a cross-resistance and three amino acid changes have additional effects on positive cross-resistance to carbofuran and permethrin.

Hypersensitivity of Somatic Mutations and Mitotic Recombinations Induced by Heterocyclic amines and Aflatoxin $B_1$ in Transgenic Drosophila (형질전환 초파리에서 Heterocyclic Amines와 Aflatoxin $B_1$에 의한 체세포 돌연변이 유발의 고감수성에 관한 연구)

  • 최영현;유미애;이원호
    • Korean journal of applied entomology
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    • v.35 no.4
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    • pp.315-320
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    • 1996
  • The effects of 2-arnino-3-methyIimidazo[4,5-fq]u inoline (IQ), 2-amino-6dimethyl-dipyrido[l,2-a;3',2'-d] imidazole (Glu-P-1) and aflatoxin B1 (AFBI) on the mitotic recombinations and somatic chromosome mutations were investigated using the transgenic Drosophila bearing a chimeric gene consisting of a promoter region of Drosophila actin 5C gene and rat DNA polymerase $. For investigating mitotic recombinations and the somatic chromosome mutations, the heterozygous (mwhl+) strain possessing or lacking transgene pol P was used. The spontaneous frequency of small mwh spots, due to deletion or nondisjunction etc., in the non-transgenic w strain and the transgenic plpol $1-130 strain was 0.351 and 0.606, respectively. The spontaneous frequency (0.063) of large mwh spots, arising mostly from somatic recombination between the centromere and the locus mwh, in the transgenic plpol $1-130 strain, was about three times higher than that (0.021) of the non-transgenic w strain. The mutant clone frequencies of two types induced by two heterocyclic mines (IQ and Glu-P-1) and AFBl in the transformant pbol PI-130 were two or three times higher than those in the host strain w. These mean that rat DNA polymerase P participates at least in the somatic chromosome mutations and mitotic recombination processes. And the present results suggest that the transgenic Drosophl!~ used in this study can be used as a hypersensitive, in vivo short-term assaying system for various environmental mutagens.

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Differentiation of Border Cells During Oogenesis in Drosophila melanogaster (노랑초파리 난자 형성과정 동안의 경계세포의 분화)

  • Gye, Myung-Chan;Cho, Kyoung-Sang;Lee, Chung-Choo
    • Development and Reproduction
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    • v.2 no.1
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    • pp.45-52
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    • 1998
  • An enhncer detector line(EDL) having P[1ArB] insertion in X chromosome with expression of reporter gene (lacZ) in the polar cells and border cell of egg chamber was established and used to monitor the differentiation and migration of border cells during the oogenesis of Drosophila. differentiation of border cell from the anterior polar follicle cells was evident in stage-9 egg chamber of EDL149 which was characterized by migration of columnar follicle cells toward posterior of egg chamber surrounding the oocyte. Migration of border cells was observed in the stage-9 and -10 egg chambers. \beta -galactosidase activities were rapidly increased during the first 4 days after eclosion, and it coincided with the timing of border cell differentiation in the ovary during adult life. Homozygote of EDL149 showed some retardation of border cell migration , resulting absence of migration of some border cells in the anterior part of egg chamber or delayed migration of some border cells in the stage-10 egg chamber. These results suggest that the P[1ArB] of EDL149 is inserted at the locus of the structural gene required for the border cell migration. In addition to the expression in egg chambers, lacZ expression was also detected in the meiotic germ cells of testis and antenna, suggesting the possible requirement of the trapped gene function in these organ. this EDL and enhancer trapped gene might be useful for the study of developmentally regulated cell migration.

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Construction of the Transgenic Drosophila melanogaster Expressing a Human Megsin Gene. (인간 신장질환 유발인자가 발현하는 형질전환 초파리 구축)

  • Goo, Tae-Won;Kwon, Ki-Sang;Kwon, O-Yu
    • Journal of Life Science
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    • v.17 no.5 s.85
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    • pp.719-722
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    • 2007
  • IgA nephropathy(IgAN) is considered to be a multifactorial disease with genetic and environmental factors contributing to its pathogenesis. The genes involved in susceptibility and progression of the disease have not yet been clearly elucidated. Megsin is an important candidate gene, predominantly expressed in glomerular mesangium and upregulated in IgAN. To understand biological function of megsin, in this work we have produced transgenic D. melanogaster fly over-expressing human megsin(actin-gal4>UAS-Megsin fly). Introduced human megsin was confirmed by RT-PCR and Western blotting, respectively. Its phenotype is melanin deficiency-abdomen and the megsin gene is stably transferred to the next generations.

Construction of Transgenic Silkworms Expressing Human Stem Cell Factor (hSCF) (인간 유래 Stem Cell Factor (hSCF) 재조합단백질이 발현되는 누에형질전환체 제작)

  • Kim, Sung-Wan;Yun, Eun-Young;Kim, Seong-Ryul;Park, Seung-Won;Kang, Seok-Woo;Kwon, O-Yu;Goo, Tae-Won
    • Journal of Life Science
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    • v.21 no.12
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    • pp.1726-1731
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    • 2011
  • Human Stem Cell Factor (hSCF) is a cytokine that binds to the c-Kit receptor and plays an important role in hematopoiesis, spermatogenesis, and melanogenesis. To produce the human Stem Cell Factor (hSCF) recombinant protein, we constructed a germline transgenic silkworm using the piggyback vector. The expression of the hSCF gene was driven by the Drosophila heat shock protein 70 (dHsp70) promoter. 3XP3 promotor-driven EGFP was used as a marker which allowed us to rapidly distinguish the transgenic silkworm. A mixture of the donor and helper vector was micro-injected into 1,020 eggs of bivoltin silkworms, Keomokjam. We obtained approximately 22 G1 broods that were EGFP-positive. The expression of the hSCF gene in the transgenic silkworm was analyzed by SDS-PAGE and immunoblotting. Also, analysis of insertion sites into the silkworm genome using inverse PCR showed that exogenous DNA was inserted into the transgenic silkworm genome. These results show that successfully constructed transgenic silkworm expresses the hSCF recombinant protein.

Study of anti-Alzheimer Activities from Ginseng Radix Rubra Water Extract by Alzheimer's Protein APP-transgenic Fly (홍삼(紅蔘) 수추출물(水抽出物)이 치매조백질(痴呆蚤白質) APP 형질전환(形質轉換) 초파리에 미치는 영향(影響))

  • Kim, Young-Jun;Kim, Jin-Heong;Yun, Jong-Hyun;Jung, Ejun-Young;Kim, Tae-Heon;Lyu, Yeong-Su;Kang, Hyung-Won
    • Journal of Oriental Neuropsychiatry
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    • v.20 no.1
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    • pp.235-247
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    • 2009
  • Objectives : Ginseng Radix Rubra water extract(RGE) has been used in vivo test for its beneficial effects on neuronal survival and neuroprotective functions, particularly in connection with APP-related dementias and Alzheimer's disease (AD). APP derived from proteolytic processing of the ${\beta}$-amyloid precursor protein (APP), including the amyloid-${\beta}$ peptide (A${\beta}$), plays a critical role in the pathogenesis of Alzheimer's dementia. Methods : We determined that RGE inhibits formation of APP, which are the behavior, and possibly causative, feature of AD. Results and Conclusions : In the cells, RGE significantly activated antiapoptosis and decreased the activity of APP-grim, a key enzyme in the apoptosis cell-signaling cascade. These results suggest that neuronal damage in AD might be due to two factors: a direct APP toxicity and multiple cellular and molecular neuroprotective mechanisms, including attenuation of apoptosis and direct inhibition of APP, underlie the neuroprotective effects of RGE.

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An Increased Intracellular Calcium Ion Concentration in Response to Dimethyl Sulfoxide Correlates with Enhanced Expression of Recombinant Human Cyclooxygenase 1 in Stably Transfected Drosophila melanogaster S2 Cells (Dimethyl sulfoxide에 의한 세포내 칼슘이온 농도 증가가 안정적으로 형질 전환된 초파리 S2 세포에서 재조합 사람 cyclooxygenase 1의 발현에 미치는 영향)

  • Chang, Kyung Hwa;Park, Jong-Hwa;Kim, Do Hyung;Chung, Ha Young;HwangBo, Jeon;Lee, Hyun Ho;Lee, Hee-Young;Shon, Dong-Hwa;Kim, Wonyong;Chung, In Sik
    • KSBB Journal
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    • v.27 no.5
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    • pp.313-318
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    • 2012
  • Dimethyl sulfoxide (DMSO) increased the intracellular calcium ion concentration in stably transfected Drosophila melanogaster S2 cells expressing recombinant cyclooxygenase 1 (COX-1). DMSO did not increase the Drosophila NOS (dNOS) transcript level in calcium chelator-treated cells. Expression of recombinant COX-1 due to DMSO was diminished in cells treated with calcium chelators or channel blockers. Our results indicate that an increased intracellular calcium ion concentration due to DMSO is associated with up-regulation of the dNOS gene, leading to enhanced expression of COX-1.