• Journal of Plant Biotechnology
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• v.46 no.3
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• pp.205-216
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• 2019

Since the RNA interference (RNAi) had been discovered in many organisms, small non-coding RNA-mediated gene silencing technology, including RNAi have been widely applied to analysis of gene function, as well as crop improvement. Despite the usefulness of RNAi technology, RNAi transgenic crops have various potential environmental risks, including off-target and non-target effects. In this study, we developed methods that can be effectively applied to environmental risk assessment of RNAi transgenic crops and verified these methods in 35S::dsRNAi_eGFP rice transgenic plant we generated. Off-target genes, which can be non-specifically suppressed by the expression of dsRNAi_eGFP, were predicted by using the published web tool, pssRNAit, and verified by comparing their expressions between wild-type (WT) and 35S::dsRNAi_eGFP transgenic rice. Also, we verified the non-target effects of the 35S:: dsRNAi_eGFP plant by evaluating horizontal and vertical transfer of small interfering RNAs (siRNAs) produced in the 35S::dsRNAi_eGFP plant into neighboring WT rice and rhizosphere microorganisms, respectively. Our results suggested that the methods we developed, could be widely applied to various RNAi transgenic crops for their environmental risk assessment.

• Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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• 2019.05a
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• pp.533-536
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• 2019

Recombinant baculoviruses are widely used to express heterologous genes in cultured insect cells. Recombinant baculoviruses can serve as gene-transfer vectors for expression of recombinant proteins in a wide range of mammalian cell types. Baculovirus system has significant benefits in view of safety, large-scale, and high level of gene expression. In this study, baculoviral vectors which were reconstructed from pOPINEneo-3C-GFP vector, were recombined with cytomegalovirus (CMV) promoter, green fluorescent protein (GFP), and p53 with NcoI and XhoI. These recombinant vectors were infected with various cells and cell lines. The baculovirus vector thus developed was analyzed by comparing the metastasis and expression of the recombinant genes with conventional vectors. These results suggest that the baculovirus vector has higher efficiency in metastasis and expression than the control vector. This work was supported by a grant from Mid-Career Researcher Program(NRF-2016R1A2B4016552) through the National Research Foundation of Korea(NRF) funded by the Ministry of Science, ICT & Future Planning(MSIP).

• Proceedings of the Korean Society of Plant Biotechnology Conference
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• 2004.05a
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• pp.49-50
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• 2004

• 한국생물공학회:학술대회논문집
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• 2005.04a
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• pp.408-411
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• 2005

The green fluorescent protein (GFP) from the jellyfish aequorea and its fluorescent homologs from Anthozoa corals have become invaluable tools for imaging of cells and tissues. In this study various fluorescent protein such as green fluorescent protein (GFP), yellow fluorescent protein (YFP) and red fluorescent protein (RFP) have been expressed in Escherichia coli. Growth of recombinant cells and production of fluorescent proteins were investigated in shake flasks. Some characteristics of fluorescent proteins was also studied.

• 한국생물공학회:학술대회논문집
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• 2003.04a
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• pp.630-633
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• 2003

Transfer of an electron from one site to another in a molecular or between molecules and/or electrodes is one of the most fundamental and ubiquitous processes in chemistry, biology and physics. In this study fusion proteins composed by green fluorescent protein(GFP) and cytochrome b562 were used in fabricating molecular array as an electron sensitizer and electron acceptor, Protein formation onto the substrate was performed by the self-assembly technique. The fusion protein film were analyzed using scanning probe microscope(SPM), Surface Plasmon Resornance(SPR) and hybrid STM/I-V. The results suggest that the proposed molecular photodiode can be used as a basic unit of the memory device.

• Journal of Microbiology and Biotechnology
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• v.16 no.3
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• pp.386-390
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• 2006

A green fluorescent protein-based Pseudomonas putida reporter was successfully constructed and shown to be capable of detecting oxidative stress. In this whole-cell reporter, the promoter of the paraquat-inducible ferredoxin-$NADP^+$ reductase (fpr) was fused to a promoterless gfp gene on a broad-host-range promoter probe vector. Pseudomonas putida KT2440 harboring this reporter plasmid exhibited an increased level of gfp expression in the presence of redox-cycling agents (paraquat and menadione), hydrogen peroxide, and potential environmental pollutant chemicals such as toluene, paint thinner, gasoline, and diesel. Induction of fpr in the presence of these chemicals was confirmed using Northern blot analysis.

• Journal of Microbiology and Biotechnology
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• v.10 no.1
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• pp.91-94
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• 2000

The suspension cells of Taxus cuspidata were transformed with Agrobacterium tumefaciens harboring binary vector pCAMBIE1302 encoding mgfp. Transient transfection efficiency was compared by using the fluoremetric measurement. The transient transfection efficiency was improved by transformation with DMSO and/or sonication treatment. Optimum conditions for DMSO and sonication treatment were 3% and 30sec, respectively. selection and maintenance of transformed cells were continued for 3 months. An insertion of the mgfp gene in transformed cells was detected by PCR and an expression of GFP confirmed by the western blot analysis.

• Journal of Microbiology and Biotechnology
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• v.10 no.2
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• pp.264-266
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• 2000

A new GFP-based T-vector for cloning of PCR products was developed by using a green fluorescent protein (GFP) as a mafker. In order to facilitate the DNA inserts, multiple restriction sites, SP6 and T7 RNA polymerase promoter sites, were introduced close to the PCR DNA insertion site of a pCRGv vector. The XcmI-digested pHNT plasmid can be used to clone a 3' A-overhanged PCR DNA amplified by Taq DNA polymerase. A potential method of easing some difficulties from its use along with its cost savings proveded by this vector are likely to lead to the replacement of other T-vectors for PCR DNA cloning.

• Bulletin of the Korean Chemical Society
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• v.26 no.3
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• pp.413-417
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• 2005

Fluorescence of green fluorescent protein mutant, 2-5 GFP is observed during denaturation by guanidine. The fluorescence intensity decreases exponentially but the fluorescence lifetime does not change during denaturation. The fluorescence lifetime of the denatured protein is shorter than that of native form. As the protein structure is modified by guanidine, solvent water molecules penetrate into the protein barrel and protonate the chromophore to quench fluorescence. Most fluorescence quenchers do not affect the fluorescence of native form but accelerate the fluorescence intensity decay during denaturation. Based on the observations, a simple model is suggested for the structural change of the protein molecule during denaturation.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 1998.07a
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• pp.59-60
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• 1998

A DNA construct containing rat T $\alpha$ 1 $\alpha$ -tuulin gene 5'-flanking sequence and GFP reporer gene was microinjected into 1-cell loach embryos. Neuron-specific FGP expression was observed in developing loach embryos and early stage fry. The results demonstrated that rat T $\alpha$ 1 $\alpha$ -tubulin gene promoter may be sufficient to specify gene expression to neurons in loach embryos. Thus, the use of GFP reporter controlled by T $\alpha$ 1 $\alpha$ -tubulin gene promoter may facilitate visualization of the dynamic processes of neural tissue development.