• Korean Journal of Plant Tissue Culture
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• v.27 no.4
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• pp.283-297
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• 2000

Seeds are an ideal repository for recombinant proteins in molecular farming applications. However, in order to use plant seeds efficiently for the production of such proteins, it is necessary to understand a number of fundamental biological properties of seeds. This includes a full understanding of promoters which function in a seed-specific manner, the subcellular targeting of the desired polypeptide and the final form in which a protein is stored. Once a biologically active protein has been deposited in a seed, it is also critical that the protein can be extracted and purified efficiently. In this review, these issues are examined critically to provide a number of approaches which may be adopted for production of recombinant proteins in plants. Particular attention is paid to the relationship between subcellular localization and protein extraction and purification. The robustness and flexibility of seed-based production is illustrated by examples close to or already in commercial production.

• Journal of Plant Biotechnology
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• v.8 no.1
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• pp.1-8
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• 2006

The differential response of soybean cultivars with or without sulfur (S) application was observed under fold conditions. Plant biomass decreased by sulfur deficiency but the reduction was less in Bragg variety about 26 % relative to the control than other ones over 45%, probably due to less reduction in loaves and pods. The photosynthetic rate of Bragg cultivar was also unaffected by the absence of sulfur application while it depressed in other lines. Soybean cultivars were compared in terms of storage protein, protein quality and biomass production by application of sulfur nutrition. The storage protein concentration tended to decrease without sulfur application in all the cultivars, however the differential response of protein quality only by 11S/7S ratio to sulfur nutrition status was observed: For instance, Bragg cultivar had higher biomass and protein production but protein quality decreased at sulfur deficiency. On the other hand, biomass and protein production in other cultivars remained louver at sulfur deficiency but protein quality differed genetically in spite of sulfur nutrition status. These results suggest that the response of soybean to sulfur nutrition is controlled by genotypic difference and sulfur supply status.

• Journal of Plant Biotechnology
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• v.7 no.2
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• pp.97-103
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• 2005

This study was carried out to obtain basic information for gene manipulation in potent edible tobacco (Nicotiana tabacum cv. TI 516). N. tabacum cv. TI 516 is a plant for a possible candidate to use as an edible vaccine, since it contains a low level of nicotine. The effective plant regeneration system through leaf disc culture was achieved using a MS basal medium supplemented with 0.1 mg $1^{-1}$ NAA and 0.5 mg $1^{-1}$ BA. In order to transform the N. tabacum cv. TI 516 with the green fluorescent protein (GFP) gene, Agrobacterium tumefaciens LBA 4404 containing the GFP gene was used. Genomic PCR confirmed the integration of the GFP gene into nuclear genome of transgenic plants. Expression of the GFP gene was identified in callus, apical meristem and root tissue of transgenic N. tabacum cv. TI 516 plants using fluorescence microscopy. Western blot analysis revealed the expression of GFP protein in the transgenic edible tobacco plants. The amount of GFP protein detected in the transgenic tobacco plants was approximately 0.16% of the total soluble plant protein (TSP), which was determined by ELISA.

• Nutrition Research and Practice
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• v.17 no.5
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• pp.1028-1041
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• 2023

BACKGROUND/OBJECTIVES: This study aimed to analyze the potential of school meals in South Korea as a sustainable tool to reduce carbon emissions by focusing on animal- vs. plant-based protein foods. MATERIALS/METHODS: By using a stratified proportional allocation method, 536 out of the 11,082 schools nationwide were selected including 21 kindergartens, 287 elementary-, 120 middle- and 108 high schools. A total of 2,680 meals served for 5 consecutive days (June 21-25, 2021) were collected. We analyzed the average serving amounts of protein foods (animal- vs. plant-based) per meal and then, calculated the estimated average amounts of carbon emission equivalents per meal by applying the conversion coefficients. The t-test and analysis of variance were used for statistical analyses (α = 0.05). RESULTS: The average serving amount of animal-based protein foods per meal was 12.5 g, which was approximately 3 times higher than that of plant-based ones (3.8 g) (P < 0.001); the Meat-group had the highest average amount of 17.0 g, followed by Egg-group (9.6 g), Fish-group (7.6 g), and Beans-and-Nuts-group (3.8 g) (P < 0.05). Specifically, pork (25.1 g) was ranked first, followed by poultry (19.6 g), processed meat products (18.0 g). The estimated average amount of carbon emission equivalents of animal-based protein foods per meal was 80.1 g CO₂e, which was approximately 31 times higher than that of plant-based ones (2.6 g CO₂e) (P < 0.001); the Meat-group had the highest average amount of 120.3 g CO₂e, followed by Fish-group (44.5 g CO₂e), Egg-group (25.9 g CO₂e), and Beans-and-Nuts-group (2.6 g CO₂e) (P < 0.05). Specifically, processed meat products (270.8 g CO₂e) were ranked first, followed by pork (91.7 g CO₂e), and processed fish products (86.6 g CO₂e). CONCLUSIONS: The results implied that school meals with plant-based alternatives could be a sustainable tool to improve carbon footprint.

• Journal of Plant Biology
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• v.20 no.4
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• pp.151-155
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• 1977

Ultrastructural changes of the protein bodies in Glycine max during early germination were studied. There were no major morphological changes in protein bodies within 3 days after the imbibition, but from the 4th day the expanse of protein bodies could be observed. In subsequent stages, the aggregation of protein bodies coalesced into a large mass and then less electron-dense material in the central part of the cell. At last it bacame highly vacuolated.

• BMB Reports
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• v.38 no.4
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• pp.440-446
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• 2005

A cDNA that was rapidly induced upon abscisic acid, cold, drought, mechanical wounding and to a lesser extent, by high salinity treatment, was isolated from Arabidopsis seedlings. It was classified as DREB subfamily member based on multiple sequence alignment and phylogenetic characterization. Since it encoded a protein with a typical ERF/AP2 DNA-binding domain and was closely related to the TINY gene, we named it TINY2. Gel retardation assay revealed that TINY2 was able to form a specific complex with the previously characterized DRE element while showed only residual affinity to the GCC box. When fused to the GAL4 DNA-binding domain, either full-length or its C-terminus functioned effectively as a trans-activator in the yeast one-hybrid assay while its N-terminus was completely inactive. Our data indicate that TINY2 could be a new member of the AP2/EREBP transcription factor family involved in activation of down-stream genes in response to environmental stress.

• The Plant Pathology Journal
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• v.32 no.5
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• pp.452-459
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• 2016

The genomic region of Grapevine fanleaf virus (GFLV) encoding the movement protein (MP) was cloned into pET21a and transformed into Escherichia coli strain BL21 (DE3) to express the protein. Induction was made with a wide range of isopropyl-${\beta}$-D-thiogalactopyranoside (IPTG) concentrations (1, 1.5, and 2 mM) each for duration of 4, 6, or 16 h. However, the highest expression level was achieved with 1 mM IPTG for 4 h. Identity of the expressed protein was confirmed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) followed by Western blotting. The expressed 41 kDa protein was purified under denaturing condition by affinity chromatography, reconfirmed by Western blotting and plate-trapped antigen enzyme-linked immunosorbent assay (PTA-ELISA) before being used as a recombinant antigen to raise polyclonal antibodies in rabbits. Purified anti-GFLV MP immunoglobulines (IgGs) and conjugated IgGs detected the expressed MP and GFLV virions in infected grapevines when used in PTA-ELISA, double antibody sandwich-ELISA, and Western blotting. This is the first report on the production of anti-GFLV MP polyclonal antibodies and application for the virus detection.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.79.2-80
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• 2003

To better understand plant defense responses against pathogen attack, we identified the transcription factor-encoding genes in the hot pepper Capsicum annuum that show altered expression patterns during the hypersensitive response raised by challenge with bacterial pathogens. One of these genes, Ca1244, was characterized further. This gene encodes a plant-specific Type IIIA - zinc finger protein that contains two Cys$_2$His$_2$zinc fingers. Ca1244 expression is rapidly and specifically induced when pepper plants are challenged with bacterial pathogens to which they are resistant. In contrast, challenge with a pathogen to which the plants are susceptible only generates weak Ca1244 expression. Ca1244 expression is also strongly induced in pepper leaves by the exogenous application of ethephon, an ethylene releasing compound. Whereas, salicylic acid and methyl jasmonate had moderate effects. Pepper protoplasts expressing a Ca1244-smGFP fusion protein showed Ca1244 localizes in the nucleus. Transgenic tobacco plants overexpressing Ca1244 driven by the CaMV 355 promoter show increased resistance to challenge with a tobacco-specific bacterial pathogen. These plants also showed constitutive upregulation of the expression of multiple defense-related genes. These observations provide the first evidence that an Type IIIA - zinc finger protein, Ca1244, plays a crucial role in the activation of the pathogen defense response in plants.

• Korean Journal Plant Pathology
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• v.11 no.4
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• pp.374-379
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• 1995

The cDNA of tobacco mosaic virus-pepper strain (TMV-P) coat protein (CP) genes were introduced into tobacco plants (Nicotiana tabacum cv. Samsun nn) using a binary Ti plasmid vector of Agrobacterium tumefaciens. these cDNAs introduced into tobacco plants were detected by polymerase chain reaction. Symptom development was distinctly suppressed in the transgenic plant introduced buy sense CP cDNA when the plant was inoculated with TMV-P, while in transgenic tobacco plants of antisense CP gene, symptom development was not suppressed as in non-transgenic plants. TMV-P concentration in the sense CP transgenic tobacco plant was decreased to 1/14 of the concentration in non-transgenic plants. Expression of the kanamycin resistance gene of these transgenic plants could be detected in the progeny.

• Journal of Plant Biotechnology
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• v.30 no.3
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• pp.281-291
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• 2003

In this review, we described the catalogues of the rice proteome which were constructed in our program, and functional characterization of some of these proteins was discussed. Mass-spectrometry is the most prevalent technique to rapidly identify a large number of proteome analysis. However, the conventional Western blotting/sequencing technique has been used in many laboratories. As a first step to efficiently construct protein cata-file in proteome analysis of major cereals, we have analyzed the N-terminal sequences of 100 rice embryo proteins and 70 wheat spike proteins separated by two-dimensional electrophoresis. Edman degradation revealed the N-terminal peptide sequences of only 31 rice proteins and 47 wheat proteins, suggesting that the rest of separated protein sports are N-terminally blocked. To efficiently determine the internal sequence of blocked proteins, we have developed a modified Cleveland peptide mapping method. Using this above method, the internal sequences of all blocked rice proteins(i, e., 69 proteins) were determined. Among these 100 rice proteins, thirty were proteins for which homologous sequence in the rice genome database could be identified. However, the rest of the proteins lacked homologous proteins. This appears to be consistent with the fact that about 45% of total rice cDNA have been deposited in the EMBL database. Also, the major proteins involved in the growth and development of rice can be identified using the proteome approach. Some of these proteins, including a calcium-binding protein that tuned out to be calreticulin, gibberellin-binding protein, which is ribulose-1.5-bisphosphate carboxylase/oxygense active in rice, and leginsulin-binding protein in soybean have functions in the signal transduction pathway. Proteomics is well suited not only to determine interaction between pairs of proteins, but also to identify multisubunit complexes. Currently, a protein-protein interaction database for plant proteins(http://genome.c.kanazawa-u.ac.jp/Y2H)could be a very useful tool for the plant research community. Also, the information thus obtained from the plant proteome would be helpful in predicting the function of the unknown proteins and would be useful be in the plant molecular breeding.