• 제목/요약/키워드: Transformed insect cells

검색결과 18건 처리시간 0.035초

Enhanced Resistance to Botrytis cinerea Mediated by Transgenic Expression of the Spider Chitinase Gene AvChit in Arabidopsis

  • Hur, Yeon-Jae;Kim, Doh-Hoon
    • International Journal of Industrial Entomology and Biomaterials
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    • 제19권2호
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    • pp.259-264
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    • 2009
  • The AvChit gene encodes for a chitinase from the spider, Araneus ventricosus. This spider, A. ventricosus, is an abundant species in Korea. Arabidopsis thaliana plants were transformed with the AvChit gene using Agrobacterium tumefaciens. Thirteen transgenic lines expressing the AvChit gene were obtained. Functional expression of the AvChit gene in transgenic Arabidopsis was confirmed by Southern, northern and western blot analysis. The AvChit cDNA was expressed as a 61 kDa polypeptide in baculovirus-infected insect Sf9 cells. AvChit protein extracted from transgenic Arabidopsis exhibited high levels of chitinase activity. Phytopathological tests showed that two transgenic Arabidopsis lines expressing the AvChit gene displayed high levels of resistance to gray mold disease (Botrytis cinerea).

친환경 살충제로서의 곤충 유충호르몬 길항제 (Insect Juvenile Hormone Antagonists as Eco-friendly Insecticides)

  • 최재영;제연호
    • 한국응용곤충학회지
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    • 제61권1호
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    • pp.101-108
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    • 2022
  • 곤충생장조절제(IGR)은 대상 해충에 대한 특이성이 높고 환경에 대한 독성이 상대적으로 낮다는 장점으로 유기합성 살충제를 효과적으로 대체할 수 있는 유망한 수단으로 여겨진다. 현재 시판되는 곤충생장조절제는 작용 기작에 따라 유충호르몬 작용제(JHA), 탈피호르몬 작용제(EA) 및 키틴 합성 저해제(CSI)의 세 가지로 구분된다. 최근 들어, 이집트숲모기의 Met과 FISC/CYC 유전자를 yeast two-hybrid system에 도입하여 유충호르몬에 의해 매개되는 Met과 FISC/CYC의 결합을 in vitro에서 구현하였으며, yeast two-hybrid β-galactosidase assay를 통하여 식물과 미생물 및 화합물 library로부터 다양한 유충호르몬 길항제(JHAN)가 분리되고 있다. 유충호르몬은 곤충의 발달, 생식, 휴면 등을 포함한 다양한 생리 작용을 조절하기 때문에, 유충호르몬 길항제는 대상 해충의 내분비 신호 전달을 방해하여 비정상적인 발달 및 유충 단계에서의 치사를 초래하며, 이는 유충호르몬 길항제가 넓은 기주 범위를 가진 살충제 개발에 효과적으로 이용될 수 있다는 것을 시사하였다. 따라서 본 논문에서는 유충호르몬 길항제의 작용점인 Met에 의해 매개되는 유충호르몬의 신호 전달 체계와 친환경 살충제로서의 유충호르몬 길항제의 전망에 대해 알아보고자 하였다.

Trichoplusia ni 세포의 apoptosis 메커니즘 규명을 위한 기초연구 (Basic Studies on the Apoptosis Mechanism of Trichoplusia ni Cell Line)

  • 이종민;양재명;이윤형;정인식
    • Applied Biological Chemistry
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    • 제44권1호
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    • pp.1-6
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    • 2001
  • 본 연구에서는 Ttichoplusia ni 세포의 apoptosis 유도 및 억제 현상의 기초연구를 수행하였다. Apoptosis 유도제로 알려진 hygromycin B에 의한 세포 성장 저해는 $200\;{\mu}/ml$의 수준에서부터 나타났고, $400\;{\mu}/ml$ hygromycin B를 처리한 세포에서는 배양 후 2일부터 DNA가 분절되어지는 것을 확인할 수 있었다. 그러나 dexamethasone과 sodium butyrate를 첨가시 세포성장은 저해되었지만 DNA 분절현상이 보이지 않아 apoptosi의 유발여부를 확인할 수 없었다. 그리고 caspase 기능억제제의 apoptosis 지연효과를 보기 위해 $200\;{\mu}/ml$ hygromycin B로 apoptosis를 유발한 상태에서 Ac-DEVD-CHO를 첨가하여 세포성장을 비교해 본 결과 이 저해제에 의해 약 36%정도 apoptosis가 억제되었음을 확인하였다. N-acetylcysteine의 경우도 apoptosis지연 효과가 있었다. Bcl_계에 속하는 anti-apoptotic 유전자의 발현연구로서 apoptosis 저해 단백질인 bcl-2 유전자를 곤충세포에 형질전환시킨 후 이 단백질이 한시적으로 발현되는 것을 western blot분석법으로 확인하였으며 apoptosis가 지연된 곤충세포주의 개발이 가능하다는 결론을 보였다.

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배추흰나비의 變態에 따른 血球細胞의 組織化學的 硏究 (Histochemical Study on the Hemocytes during Metamorphosis)

  • Kim, Chang-Whan;Kim, Woo-Kap;Kim, Yong-Kuk
    • 한국동물학회지
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    • 제11권4호
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    • pp.103-117
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    • 1968
  • 1. 變態時期에 따른 배추흰나비(Pieris rapae)의 血球를 組織化學的方法 및 autoradiographic method 에 의거하여 硏究하였다. 2.幼蟲, 전용 및 용기에서 形態의變化와 細胞內容物을 土臺로 血球細胞를 prohemocyte, plasmatocyte, podocyte, granular hemocyte, spherule cell 및 oenocytoid 의 6型으로 分類했다. 3, 幼蟲基에서 용기까지의 血球를 組織化學的으로 硏究하고 血球細胞內의 蛋白質, 核酸, 多糖類 및 脂質을 檢出하였으며, 成長 및 變態에 따른 그들 物質의 消長을 밝혔다, 4. Granular hemocyte 와 spherule cell 의 大部分의 顆粒은 glycogen, neutral mucopolysaccharide, mucoprotein 으로 되었고 neutral fat와 phospholipid 의 顆粒도 나타났다, 5. 細胞分裂像(mitotic figure) 과 tritium-thymidine 攝取세포 (DNA 合成細胞)를 幼蟲 2齡부터 5齡까지의 各種 未分化血球에서 觀察하였고, 모든型의 血球가 prohemocyte에 起源함을 밝혔다. 6. Plasmatocyte 와 podocyte 의 細胞質突起가 전용기 와 용기에 매우 길게 伸張함을 보아 vermiform cell 은 plasmatocyte 의 더 分化한 型이라고 생각된다.

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Metabolic Engineering of Indole Glucosinolates in Chinese Cabbage Plants by Expression of Arabidopsis CYP79B2, CYP79B3, and CYP83B1

  • Zang, Yun-Xiang;Lim, Myung-Ho;Park, Beom-Seok;Hong, Seung-Beom;Kim, Doo Hwan
    • Molecules and Cells
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    • 제25권2호
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    • pp.231-241
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    • 2008
  • Indole glucosinolates (IG) play important roles in plant defense, plant-insect interactions, and stress responses in plants. In an attempt to metabolically engineer the IG pathway flux in Chinese cabbage, three important Arabidopsis cDNAs, CYP79B2, CYP79B3, and CYP83B1, were introduced into Chinese cabbage by Agrobacterium-mediated transformation. Overexpression of CYP79B3 or CYP83B1 did not affect IG accumulation levels, and overexpression of CYP79B2 or CYP79B3 prevented the transformed callus from being regenerated, displaying the phenotype of indole-3-acetic acid (IAA) overproduction. However, when CYP83B1 was overexpressed together with CYP79B2 and/or CYP79B3, the transformed calli were regenerated into whole plants that accumulated higher levels of glucobrassicin, 4-hydroxy glucobrassicin, and 4-methoxy glucobrassicin than wild-type controls. This result suggests that the flux in Chinese cabbage is predominantly channeled into IAA biosynthesis so that coordinate expression of the two consecutive enzymes is needed to divert the flux into IG biosynthesis. With regard to IG accumulation, overexpression of all three cDNAs was no better than overexpression of the two cDNAs. The content of neoglucobrassicin remained unchanged in all transgenic plants. Although glucobrassicin was most directly affected by overexpression of the transgenes, elevated levels of the parent IG, glucobrassicin, were not always accompanied by increases in 4-hydroxy and 4-methoxy glucobrassicin. However, one transgenic line producing about 8-fold increased glucobrassicin also accumulated at least 2.5 fold more 4-hydroxy and 4-methoxy glucobrassicin. This implies that a large glucobrassicin pool exceeding some threshold level drives the flux into the side chain modification pathway. Aliphatic glucosinolate content was not affected in any of the transgenic plants.

Gene functional analysis of Harmonia axyridis by in vitro transcription

  • Park, Sang-Eun;Youn, Young-Nam
    • 농업과학연구
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    • 제46권3호
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    • pp.471-488
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    • 2019
  • Random genes were screened in two transforming ways to investigate the new genes of a ladybug using the Harmonia axyridis cDNA library stock cell cloned in the LITMUS 28i vector in a previous study. Phenotypic variation was observed after injection of the synthesized double-stranded RNA through the in vitro transcription process. The cDNA library of H. axyridis was transformed into E. coli $DH5{\alpha}$ and 10B competent cells by heat shock. Analysis of the nucleotide sequences of the 42 clones with the insert DNAs revealed that 21 clones were homologous with the genes of insects, and only one clone had a gene from H. axyridis. Thirteen of the 21 insect genes were homologous with genes from coleopteran insects. Fourteen genes were selected, which were identified by the gene screening results, and were synthesized as double-stranded RNA through in vitro transcription. One microgram of the synthesized double-stranded RNA between segments T1 and T2 were injected using a syringe into each anesthetized fourth larvae which were under 2 days old. As a result, a phenotypic variation appeared in the larva injected with the two genes. While the eggs of H. axyridis injected with distilled water hatched out three days after oviposition, the eggs of H. axyridis injected with dsHma 06 did not hatch but become shrivel a week after oviposition. Most of the H. axyridis injected with dsHma 08 died and were unable to complete the pupation or eclosion during ecdysis.