• Title/Summary/Keyword: CaMV

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Introduction of PMT(Putrescine N-Methyltransferase) Antisense RNA to Tobacco Plants (담배식물체에 PMT(Putrescine N-Methyltransferase) antisense RNA유전자 도입)

  • 김선원;박성원;이정헌;이청호;류명현;복진영;김도훈;최순용
    • Journal of the Korean Society of Tobacco Science
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    • v.25 no.1
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    • pp.12-19
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    • 2003
  • Transgenic tobacco plants were selected by using the transformation of putrescine N-methyltransferase(PMT) gene, the key enzyme in diverting polyamine metabolism towards the biosynthesis of nicotine. PMT was fused in reverse orientation to the CaMV 35S promoter of the plant expression vector pBTEX(pPAB3) to produce tobacco plants of low nicotine content. To compare nicotine content, only pBTEX vector and PMT gene which was fused in forward orientation to the CaMV 35S promoter(pPAB2) were also transformed to the leaf tobacco plants(Nicotiana tabacum cv. NC82 and N. tabacum cv. Br2l). The presence of sense- and antisense-PMT gene, and pBTEX vector in the transgenic plant was confirmed by genomic PCR.

Impact of SV40 T antigen on two multiple fission microalgae species Scenedesmus quadricauda and Chlorella vulgaris

  • Gomaa, Ahmed E.;Yang, Seung Hwan
    • International journal of advanced smart convergence
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    • v.7 no.1
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    • pp.48-63
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    • 2018
  • The combination of Simian Virus40 (SV40)'s large T antigen with its replication origin is commonly used in molecular studies to enhance the expression of heterogeneous genes through multiplying the plasmid copy number. There are no reports related to the impact of the SV40 T antigen on plant, multiple fissional, cell-type. This study explores the response of two multiple-fission microalgal cells, Scenedesmus quadricauda and Chlorella vulgaris, to the expression of the T-antigen, with aim of applying SV40 T-antigen to increase the expression efficiency of foreign genes in the two species. Different levels of low-expression have been constructed to control the expression of SV40 T antigen using three heterogenous promoters (NOS, CaMV35S, and CMV). Chlorella cultures showed slowdown in the growth rate for samples harboring the T antigen under the control of CaMV35S and CMV promoters, unlike Scenedesmus cultures which showed no significant difference between samples and could have silenced the expression.

Expression of Glutathione Reductase Gene in Transgenic Tobacco Plant (형질전환 담배 식물체에서 Glutathione Reductase 유전자의 발현)

  • 이효신;조진기
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.2
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    • pp.87-90
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    • 2001
  • BcGRl gene encoding cytosolic glutathione reductase of Chinese cabbage (Brassica campestris var. Pekinensis cv. Seoul) was placed under the control of the CaMV 35S promoter and introduced into tobacco (Nicotiana tabacum L. cv. Samsun) via Agrobacterium-mediated transformation. T$_{0}$ 32 independent plants transformed with BcGRl gene were selected with kanamycin and they were confirmed by polymerase chain reaction (PCR) and Southern blot analysis. Northern blot analysis revealed that the constitutive expression of BcGRl gene and there was no relationship between the copy number of introduced gene and the levels of BcGRl transcripts.

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Proteomic analysis of dehydroascorbate reductase transgenic potato plants (Dehydroascorbate reductase 과발현 형질전환 감자 식물체의 단백질체 분석)

  • Han, Eun-Heui;Goo, Young-Min;Kim, Yun-Hee;Lee, Shin-Woo
    • Journal of Plant Biotechnology
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    • v.43 no.2
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    • pp.223-230
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    • 2016
  • Ascorbic acid (AsA) is a strong antioxidant/reducing agent that can be converted to dehydroascorbate (DHA) by oxidation in plants. DHA, a very short-lived chemical, is recycled to AsA by dehydroascorbate reductase (DHAR). Previously, DHAR cDNA was isolated from the hairy roots of the sesame plant, and DHAR-overexpressing transgenic potato plants were generated under the control of the CaMV35S promoter (CaMV35S::DHAR). An increase in transgene expression and ascorbate levels were observed in the transgenic plants. In the present study, proteomic analysis revealed that transgenic plants not only accumulated DHAR in their cells, but also induced several other antioxidant enzyme-related proteins during plant growth. These results suggest that DHAR is important for stress tolerance via induction of antioxidant proteins, and could improve stress tolerance in transgenic potato plants.

Uptake and Expression of Foreign Genes Using Seed-Derived Embryos of Rice (벼 종자 유래 배에서 외래유전자의 도입과 발현)

  • 정구흥
    • Journal of Plant Biology
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    • v.37 no.1
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    • pp.77-83
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    • 1994
  • DNA uptake in dry embryos of rice by DNA imbibition was detected by monitoring the expression of chimeric vectors. The selective markers of expression vectors used were ${\beta}-glucuronidase$ ronidase (GUS) and hygromycin phosphotransferase (HPT) genes under the control of CaMV35 S promoter. Frequency of transient expression of the foreign gene was generally 30-50% varying according to the types of vectors and rice cultivars. Dot blot analysis and DNA sequence analysis of inverse polymerase chain reaction products showed that selected rice in hygromycin B (HmB) medium had HPT gene and CaMV35S promoter DNA sequence in genomic DNA of rice. To investigate what ratio of rice having two marker genes simultaneously as rice embryos imbibed the vector DNA having two HPT and GUS gene, transform ants selected in lImB medium were subjected to PCR for GUS gene. It was shown that about 90 percentage of surviving ones in HmB medium had GUS gene.S gene.

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Expression of AmA1 Gene Encoding Storage Protein of Amaranthus in Transgenic Tobacco (형질전환 담배에서 Amaranthus 저장단백질인 AmA1 유전자의 발현)

  • 김태금;김영숙;권태호
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.3
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    • pp.169-173
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    • 2000
  • A 1,183bp cDNA, AmA1, encoding the seed storage protein of Amaranthus hypochondriacus was isolated by reverse transcriptase-polymerase chain reaction (RT-PCR) and characterized. AmA1 gene was subcloned into plant binary vector under Cauliflower Mosaic Virus (CaMV) 35S promoter and nopaline synthase terminator (3'NOS). The recombinant binary vector was used to transform Nicotiana tabacum using Agrobacterium tumefacien -mediated transformation procedure. Shoots were induced on MS medium with 0.1 mg/L NAA, 1.0 mg/L BA, 100 mg/L kanamycin and 250 mg/L cefotaxime. Transgenic plants were selected on rooting medium based on MS medium containing 200 mg/L kanamycin and 250 mg/L cefotaxime without phytoregulators. The presence of AmA1 gene in the transgenic plants was confirmed by PCR followed by DNA hybridization. The expression of AmA1 gene in the transgenic plant was observed by RT-PCR method.

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Selection of Transgenic Potato Plants Expressing NDP Kinase 2 Gene with Enhanced Tolerance to Oxidative Stress (NDP Kinase 2 유전자를 도입한 산화스트레스 내성 형질전환 감자의 선발)

  • Li, Tang;Kwon, Suk-Yoon;Yun, Dae-Jin;Kwak, Sang-Soo;Lee, Haeug-Soon
    • Journal of Plant Biotechnology
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    • v.31 no.3
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    • pp.191-195
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    • 2004
  • Arabidopsis NDPK2 (AtNDPK2) is a key singaling component that regulate cellular redox state and known to enhance multiple stress tolerance when over-expressed in Arabidopsis plant (Moon et al. 2003). In order to develop transgenic potato plants with enhanced tolerance to multiple stresses, we placed an AtNDPK2 cDNA under the control of a stress-inducible SWPA2 promoter or enhanced CaMV 35S promoter. Transgenic potato plants (cv. Superior and Atlantic) were generated using an Agrobacterium-mediated transformation system and selected on MS medium containing 100 mg/L kanamycin. Genomic Southern blot analysis confirmed the incorporation of AtNDPK2 cDNA into the potato genome. When potato leaf discs were treated with methyl viologen (MV) at 10 $\mu$M, transgenic plants showed higher tolerance to MV than non-transgenic or vector-transformed plants. The NDPK2 transgenic potato plants will be further used for analysis of stress-tolerance to multiple environmental stresses.

Characterization and RT-PCR Detection of Turnip Mosaic Virus Isolated from Chinese Cabbage in Korea (배추에서 분리한 순무 모자이크 바이러스의 특성 및 역전사 중합효소 연쇄반응법(RT-PCR)을 이용한 검정)

  • 박원목;최설란;김수중;최승국;류기현
    • Korean Journal Plant Pathology
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    • v.14 no.3
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    • pp.223-228
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    • 1998
  • Turnip mosaic virus)TuMV-Ca) was isolated from a Chinese cabbage showing severe mosaic and black necrotic spots symptoms in Korea. The virus was identified as a strain of TuMV by its host range test, particle morphology, serology, double stranded RNA analysis. For detection of the virus, reverse transcription and polymerase chain reaction(RT-PCR) was performed with a set of 18-mer TuMV-specific primers to amplify a 876 bp DNA fragment The virus was rapidly detected from total nucleic acids of virus infected tissues as well as native viral RNA of purified virion particles by RT-PCR. Detection limit of the viral RNA by RT-PCR was 10 fg.

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