• Journal of Plant Biotechnology
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• 제45권4호
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• pp.322-327
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• 2018

Post-translational modification of proteins regulates signaling cascades in eukaryotic system, including plants. Among these modifications, phosphorylation plays an important role in modulating the functional properties of proteins. Plants perceive environmental cues that directly affect the phosphorylation status of many target proteins. To determine the effect of environmentally induced phosphorylation in plants, in vivo methods must be developed. Various in vitro methods are available but, unlike in animals, there is no optimized methodology for detecting protein phosphorylation in planta. Therefore, in this study, a robust, and easy to handle Phos-Tag Mobility Shift Assay (PTMSA) is developed for the in vivo detection of protein phosphorylation in plants by empirical optimization of methods previously developed for animals. Initially, the detection of the phosphorylation status of target proteins using protocols directly adapted from animals failed. Therefore, we optimized the steps in the protocol, from protein migration to the transfer of proteins to PVDF membrane. Supplementing the electrophoresis running buffer with 5mM $NaHSO_3$ solved most of the problems in protein migration and transfer. The optimization of a fast and robust protocol that efficiently detects the phosphorylation status of plant proteins was successful. This protocol will be a valuable tool for plant scientists interested in the study of protein phosphorylation.

• 대한수의학회지
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• 제52권1호
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• pp.25-31
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• 2012

Brucella (B.) canis is mainly transmitted by direct or indirect contact with aborted fetuses and placenta. It's also known to be able to infect human, which likely results in providing veterinarians and companion animal owners for infectious risk. To develop diagnostic ELISA, we cloned and expressed rp1L gene of B. canis, which encodes the ribosomal protein L7/L12. Using this purified recombinant protein, indirect-ELISA (iELISA) was evaluated using 78 positive and 44 negative sera. The sensitivity and the specificity of iELISA were 94% and 89%, respectively. The results indicated that indirect-ELISA using recombinant ribosomal protein L7/L12 may be useful for diagnosis of canine brucellosis.

• BMB Reports
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• 제37권1호
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• pp.133-138
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• 2004

Proteomics is a leading technology for the high-throughput analysis of proteins on a genome-wide scale. With the completion of genome sequencing projects and the development of analytical methods for protein characterization, proteomics has become a major field of functional genomics. The initial objective of proteomics was the large-scale identification of all protein species in a cell or tissue. The applications are currently being extended to analyze various functional aspects of proteins such as post-translational modifications, protein-protein interactions, activities and structures. Whereas the proteomics research is quite advanced in animals and yeast as well as Escherichia coli, plant proteomics is only at the initial phase. Major studies of plant proteomics have been reported on subcellular proteomes and protein complexes (e.g. proteins in the plasma membranes, chloroplasts, mitochondria and nuclei). Here several plant proteomics studies will be presented, followed by a recent work using multidimensional protein identification technology (MudPIT).

• 식물조직배양학회지
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• 제27권5호
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• pp.387-393
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• 2000

The process by which Agrobacterium tumefaciens genetically transforms plants involves a complex series of reactions communicated between the pathogen and the plants. To identify plant factors involved in agrobacterium-mediated plant transformation, a large number of T-DNA inserted Arabidopsis thaliana mutant lines were investigated for susceptibility to Agrobacterium infection by using an in vitro root inoculation assay. Based on the phenotype of tumorigenesis, twelve T-DNA inserted Arabidopsis mutants(rat) that were resistant to Agrobacterium transformation were found. Three mutants, rat1, rat3, and rat4 were characterized in detail. They showed low transient GUS activity and very low stable transformation efficiency compared to the wild-type plant. The resistance phenotype of rat1 and rats resulted from decreased attachment of Agrobacterium tumefaciens to inoculated root explants. They may be deficient in plant actors that are necessary for bacterial attachment to plant cells. The disrupted genes in rat1, rat3, and rat4 mutants were coding a arabinogalactan protein, a likely cell wall protein and a cellulose synthase-like protein, respectively.

• Journal of Plant Biotechnology
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• 제2권1호
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• pp.1-12
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• 2000

• Journal of Photoscience
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• 제7권3호
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• pp.123-128
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• 2000

Phosphorylation of cellular proteins is a key regulatory mehanism for signal transduction pathway in living cells. Phytochrome, a red/far-red light photoreceptor in plants, is known to employ protein phosphorylation for its light signaling, although its detauked mechanism is still ambiguous. This study is intended to identify the phosphoproteins and protein kinases that are regulated by phytochrome, by employing transgenic rice seedlings that overexpress Arabidopsis phytochrome A. Red light stimulated phsophorylation of a 70 kDa protein and far-red light negated the effect. The red light induced phosphotylation of the 70 kDa protein was strongly activated by heparin and inhibited by poly-L-lysine, suggesting that the 70 kDa protein phosphorylating kinase belongs to GSK-3/SHAGGY protein kinase that has functional roles in establishing cell fate and pattern formation in Drosophila. Taken together with the fact that phytochrome controls plant development, these results may suggest that a GSK-3/SHAGGY-related protein kinase in plant(ASK) is likely to be involved in phytochrome signal transduction.

• 한국광과학회:학술대회논문집
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• 한국광과학회 2003년도 학술대회지
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• pp.58-68
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• 2003

• Journal of Nutrition and Health
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• 제22권4호
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• pp.223-236
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• 1989

To evaluate the differences of the levels and sources of protein intake human protein metabolism, an 26-day metabolic balance study was conducted in 10 healthy Korean adult females. In the pre-study, the subjects recorded their own diets for 3 days. The metabolic balance study consisted of 6-day adaptation period, 10-day moderate protein period(60-65g/d) and 10-day high protein period(90-95g/d). During the moderate and high protein period, 5 subjects were fed the higher animal protein meals and the other 5 subjects were fed the high plant protein meals. Body weight, nitrogen balance and blood chemistries were monitored through out the study. The urine volume were sighificantly larger in the animal protein group and, the dietary fiber and fecal weights were significantly heavier in the plant protein diet group. But no statistically significant differences were found between the two dietary groups in apparent nitrogen digestability, urinary nitrogen excretion and nitrogen balance. Body weight, serum protein, albumin and HDL-cholesterol levels were not changed, but serum total cholesterol level in the animal protein diet group was elevated significantly from 143.8mg/dl on moderate potein diet to 173.0mg/dl on high proetin diet. In conclusion, from the observation of this short-term N balance study, plant diet on the adequate level of calorie and protein intake had almost the same effect of animal protein diet for protein maintenace in adults.

• Journal of Plant Biology
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• 제36권1호
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• pp.83-90
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• 1993

As a part of the study on the relation between exogenous polyamines and various components necessary for protein biosynthesis in the germinating maize seeds, the effects of the polyamines on protein biosynthesis and irbosome activity were investigated. The protein biosynthesis activity by S-30 containing all components necessary for protein biosynthesis was increased by exogenous polyamines, spermidine, spermine and putrescine. As the concentration of polyamine treated was increased, the optimal Mg2+ concentration of in vitro poly U-dependent protein synthesis system was gradually reduced. However, the optimal Mg2+ concentration of poly U-dependent system containing optimal polyamine was 10 mM regardless of the sort of polyamine. It could be infered that polyamines play an important part in protein biosynthesis in the higher plant, and that the role of polyamines take partially the place of Mg2+ action. The activities of ribosome and S-100 in protein biosynthesis were increased by 46.7% and 17.7% with spermidine, and by 44.1% and 16.2% with spermine, and by 29.1% and 19.3% with putrescine. It could be concluded that the increase of protein biosynthesis by polyamines in mainly owing to the ribosome activation.

• Journal of Nutrition and Health
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• 제24권2호
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• pp.124-131
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• 1991

식이 단백질 섭취수준 및 동·식물성 단백질 급원이 칼슘 및 인대사에 미치는 영향을 검토하기 위하여 10명의 한국성인 연성을 대상으로 26일간의 통제 식이실험을 실시하였다. 대사실험 연구는 6일간의 적응기간과 10일간의 중단백식이(60g 단백질 545mg Ca)와 10일간의 고단백질 식이(90g 단백질 575mg Ca)로 구성되었다 중·고단백 식이기간 동안 동물성 단백질(75% 동물성 단백질)과 식물성 단백질(75% 식물성 단백질) 식이군으로 나누어 같은 식이에서 중단백식이 후 고단백식이를 섭취토록 하였고 실험식이 마지막 4일간은 각 2명에게 300mg 칼슘을 보충시켰다. 칼슘 흡수율은 단백질 섭취량의 증가시 동물성 단백질 식이군에서 유의적으로 증가하였다, 칼슘 보충은 급원에 관계없이 칼슘 흡수를 증가시켰다. 중단백 식이기간 동안 동물성과 단백질 식이에서 칼슘흡수율은 약 30% 이었으며 고단백질식이 기간 동안 동물성 단백질에서 46% 식물성 단백질에서 37% 이었다. 칼슘을 보충시켰을 때 중단백 식이기간동안 칼슘 흡수율은 약 46% 이었으며 고단백 식이군에서 53%이었다 뇨중 칼슘배설은 단백질 섭취수준에는 영향을 받지 않았으나 동물성 단백질 식이에서 식물성 단백질 식이보다 칼슘배설양이 많았다. 칼슘 균형을 단백질 섭취증가와 칼슘 보충으로 호전되었다. 인의 흡수는 단백질 섭취수준에는 영향을 받지않았으며 인 섭취량에 따라 증가하였다. 인 흡수율은 동물성 단백질 식이군에서 약 77∼81% 로 식물성 단백질 식이군에서의 55∼65% 보다 높았다. 이상의 결과로 보아 단백질 섭취수준 60g에서 90g으로의 증가와 칼슘보충은 뇨중 칼슘 배설에는 영향을 주지않으며 칼슘 흡수를 증진시켜 칼슘균형을 호전시키는 것으로 나타났다.