• 한국자원식물학회지
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• 제10권3호
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• pp.221-226
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• 1997

Improvement of seed protein quality might be an essential issus in soybean and would give more profit directly to both farmers and users. This study was carried out to investigate the effects of reduced-S form(s) on seed storage protein components in soybean during seed filling stages. The reduced-S forms during seed fill were sodium thiosulfate, sodium sulfite, sodium sulfide, thioaceteat, $\beta$-mercaptoethanol, thiourea, thiamine-HCI, L-cysteine, L-cystine, and L-methionine. Seed storage protein concentration did not appear to be affected by any reduced-S forms. However, glycinin and $\beta$-conglycinin concentration seemed to be changed greatly by L-methionine. This resulted in the increase in the 11S/7S ratio(3.58). Among the $\beta$-conglycinin, $\beta$-subunit was not accumulated at all. $\alpha$-subunit concentration appeared to be decreased and $\alpha'$-subunit concentration was not altered in comparison with sulfate control. Also, $\beta$-conglycine concentration, especially $\beta$-subunit concentration, tended to be decreased with L-cystine treatment, resulting in an increase in the 11S/7S ratio(1.83). The glycinin concentration tended to be increased at the expense of the decrease in the $\beta$-conglycinin concentration. Therefore, it is suggested that enhancing soybean protein quality would be achieved by improving metabolic pathways of S assimilation in soybean plants during seed filling period under sulfate-sufficient condition.

• Applied Biological Chemistry
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• 제32권1호
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• pp.64-72
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• 1989

Near full length cDNA clones encoding the rice seed storage protein, prolamine, were isolated and divided into two homology classes based on cross-hybridization and DNA sequencing analysis. These cDNA clones contain a single open reading frame encoding a putative rice prolamine precursor(M.W.=17,200) possessing atypical 14 amino acid signal peptide. Clones of these two homology classes diverge mainly by insertions/deletions of short nucleotide stretches and point mutations. The deduced primary structures of both types of prolamine polypeptides are devoid of any major tandem repetitive sequences, a feature prevalent in other cereal prolamines. No significant homology teas detected between the rice prolamine and other cereal prolamines, indicating that the rice gene evolved from a different ancestor that gave rise to other cereal prolamine genes. Developing wheat and rice endosperms were examined using ultrathin sections prepared from tissues harvested at various days after flowering. By immunocytochemical localization techniques, wheat prolamines are localized within vesicles from Golgi apparatus and in homogeneous regions of protein bodies. The involvement of the goli apparatus in the packaging of wheat prolamines into protein bodies indicates a pathway which differs from the mode of other cereal prolamines and resembles the mechanism employed for the storage of rice glutelin and legume globulins.

• Plant Resources
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• 제1권1호
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• pp.6-12
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• 1998

The Fp1 gene, originally isolated from red pepper seedlings, encode the iron storage protein, and have a high homology with ferritin genes at DNA and amino acid level. In order to determine ferritin protein expression in vegetative tissue. Fp1 gene was constructed in plant expression vector(PIG12IHm) and introduced in red pepper(var. Bukang, Chungyang and Kalag-Kimjang 2) via Agrobacterium tumefaciensmediated transformation. After selection on MS media containing Kanamycin(Km), putatively selected transformants were confirmed by amplification of selectable marker gene(Fp1 and NPII) by polymerase chain reaction. Northern blot showed that transcripts of Fp1 gene were detected in mature leaves of the plants. In A6, A7 and A8 and A14 of transgenic plants, transcript of Fp1 gene was increased seven-fold to eight-fold than other transgenic plants. Also the proteins obtained from leaves of transgenic plants were immunologically detected by Western blot using rabbit anti-ferritin polyclonal antibody. The expression protein appeared as strong band of apparent mass of 23.5kDa. suggesting the iron accumulation in transgenic red pepper plants.

• Journal of Microbiology and Biotechnology
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• 제16권6호
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• pp.901-910
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• 2006

Poly(3-hydroxybutyrate) [P(3HB)] is a microbial polyester intracellularly accumulated as distinct granules in numerous microorganisms as an energy and carbon storage material. Recombinant Escherichia coli harboring the heterologous P(3HB) biosynthesis genes accumulates large amounts of P(3HB) granules, yet the granule-associated proteins have not been identified. Therefore, this study reports on an analysis of the P(3HB) granule-associated proteome in recombinant E. coli. Fiye proteins out of 7 spots identified were found to be involved in functions of translation, heat-stress responses, and P(3HB) biosynthesis. Two of the major granule-associated proteins, IbpA/B, which are already known to bind to recombinant proteins forming inclusion bodies in E. coli, were further analyzed. Immunoblotting and immunoelectron microscopic studies with IbpA/B antibodies clearly demonstrated the binding and localization of IbpA/B to P(3HB) granules. IbpA/B seemed to play an important role in recombinant E. coli producing P(3HB) by stabilizing the interface between the hydrophobic P(3HB) granules and the hydrophilic cytoplasm. Thus, IbpA/B were found to act like phasins in recombinant E. coli, as they are the major proteins bound to the P(3HB) granules, affect the morphology of the granules, and reduce the amount of cytosolic proteins bound to the P(3HB) granules.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2010년도 정기총회 및 춘계학술발표회
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• pp.14-14
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• 2010

Plastid proteomics are essential organelles present in virtually all cells in plants and green algae. Plastids are responsible for the synthesis and storage of key molecules required for the basic architecture and functions of plant cells. The proteome of plastid, and in particular of chloroplast, have received significant amounts of attention in recent years. Various fractionation and mass spectrometry (MS) techniques have been applied to catalogue the chloroplast proteome and its sub-organelles compartments. To better understanding the function of the lumenal sub-organelles within the thylakoid network, we have carried out a systematical analysis and identification of the lumenal proteins in the thylakoid of wheat by using Tricine-SDS-PAGE, and LTQ-ESI-FTICR mass spectrometry followed by SWISS-PROT database searching. We isolation and fractionation these membrane from fully developed wheat leaves using a combination of differential and gradient centrifugation couple to high speed ultra-centrifuge. After collecting all proteins to eliminate possible same proteins, we estimated that there are 407 different proteins including chloroplast, chloroplast stroma, lumenal, and thylakoid membrane proteins excluding 20 proteins, which were identified in nucleus, cytoplasm and mitochondria. A combination of these three programs (PSORT, TargetP, TMHMM, and TOPPRED) was found to provide a useful tool for evaluating chloroplast localization, transit peptide, transmembranes, and also could reveal possible alternative processing sites and dual targeting. Finally, we report also sub-cellular location specific protein interaction network using Cytoscape software, which provides further insight into the biochemical pathways of photosynthesis. The present work helps understanding photosynthesis process in wheat at the molecular level and provides a new overview of the biochemical machinery of the thylakoid in wheat.

• Applied Microscopy
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• 제29권4호
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• pp.499-509
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• 1999

완두 종자에 축적되는 저장물질은 주로 전분과 단백질로서 이러한 저장물질 때문에 고정이나 전자현미경 관찰시료를 제작하기가 쉽지 않다. 따라서 자엽을 얇게 절편을 만들고 효소를 사용하여 단일세포로 분리한 다음 고정하여 관찰하였다. 완두의 저장단백질이 축적되는 단백질 저장 액포는 종자발달의 이른 시기에 기존의 액포를 둘러싸고 발달하게 되므로서 액포는 수축되고 단백질 저장 액포는 점점 발달하여 그 가장자리에 단백질 덩어리가 축적되게 된다. 이와는 별도로 종자발달의 이른 시기에 조면소포체의 내강에 전자밀도가 높은 단백질이 축적되기 시작하여 늦은 시기에 이 소포체의 끝이 부풀어서 구형의 단백과립으로 발달하였다. 완두종자의 저장단백질은 주로 vicilin과 legumin으로서 단백과립에 대한 면역세포화학적 방법으로 확인한 결과 vicilin은 세포질에 발달된 작은 단백과립과 단백질 저장액포의 가장자리에 축적된 단백질 덩어리에 모두 반응하였으나 legumin은 세포질의 단백과립에만 반응하였다. 또한 소포체에 존재하는 단백질인 Bip은 단백질 저장액포에 축적된 단백질 덩어리의 안쪽 가장자리에만 반응하였다. 이는 단백질이 활발하게 축적되고있는 시기에 특징적으로 작용하는 Bip의 기능과 관련되는 것으로 사료된다.

• Asian-Australasian Journal of Animal Sciences
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• 제32권6호
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• pp.881-890
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• 2019

Objective: A study was conducted to develop non-dairy creamer analogs/mimics using egg white, egg yolk, soy protein and their combinations, and their nutrient content, shelf-life and flavor acceptability were compared. Methods: Spray dried egg white, egg yolk, and soy protein isolate were purchased from manufacturers and used for the formulae. Results: The protein contents of the non-dairy creamer analogs/mimics were about 8.5% as calculated. The amounts of oleic and linoleic acid content increased as the amount of yolk increased in the formula, but the increases of polyunsaturated fatty acids were <0.5% of total fat. Addition of egg yolk to the formula increased choline and lutein content in the products, but the amounts were <0.4 mg/g for choline and $4{\mu}g/g$ for lutein. The lutein in the products continued to decrease over the storage time, and only about 15% to 20% of the 0-month amounts were left after 3 months of storage. Although the thiobarbituric acid reactive substances values of the spray-dried non-dairy creamer analogs/mimics increased as storage time increased, the values were still low. Yellowness, darkness, and egg flavor/odor of the non-dairy creamer analogs/mimics increased as the amount of egg yolk in the formula increased. The overall acceptability of the non-dairy creamer analogs/mimics was closely related to the intensity of egg flavor/odor, but storage improved their overall acceptance because most of the off-odor volatiles disappeared during the storage. Water temperature was the most important parameter in dissolving spray-dried non-dairy creamer analogs/mimics, and $55^{\circ}C$ to $75^{\circ}C$ was the optimal water temperature conditions to dissolve them. Conclusion: Higher amounts of yolk and soy protein combinations in place of egg white reduced the cost of the products significantly and those products contained better and balanced nutrients than the commercial coffee creamers. However, off-flavor and solubility were two important issues in the products.

• 한국동물생명공학회지
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• 제34권3호
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• pp.190-196
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• 2019

Tight junctions are constituents of the blood-epididymis barrier that play roles in regulating the unidirectional transcellular transport of ions, water, and solutes to maintain optimal conditions for sperm maturation and storage. Claudin 1 (Cldn1) and 4 (Cldn4) are known as tight junction proteins and are expressed in the basolateral membranes as well as tight junctions in the epididymis of rodents. Here, we examined the expression and localization of Cldn1 and 4 to determine the function of these proteins in the pig epididymis. Cldn1 was highly expressed in the basolateral membrane of epithelial cells in the caput and corpus regions of the epididymis. In the cauda region, however, Cldn1 labeling was significantly decreased in the basolateral membrane of epithelial cells. In contrast, labeling indicated that Cldn4 was expressed in the basolateral membrane in the cauda region of the epididymis and was present at punctate reactive sites in the caput and corpus regions. However, in no region of the epididymis did we detect colocalization of Cldn1 and 4 with labeled ZO-1, the distribution of which is restricted to the tight junctions. Our results indicate that Cldn1 and 4 were region-specifically expressed in the pig epididymis but not present in the tight junctions of epididymal epithelium. In addition, reciprocal regulation in specific regions of the epididymis between Cldn1 and 4 may play an important role in generating an optimal luminal environment for sperm maturation and storage in the pig epididymis.

• Asian-Australasian Journal of Animal Sciences
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• 제18권3호
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• pp.427-432
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• 2005

The purpose of this study was performed as model study using four animals to investigate the correction between the changes in Differential Scanning Calorimetry thermogram of muscle proteins during storage and meat freshness. M. longissimus dorsi of pork was obtained immediately after slaughter and chilled/stored at either $-2^{\circ}C$ or $25^{\circ}C$ for up to 96 h for analyses. DSC thermograms were determined and compared with pH values, ATP-related compounds, K-values, volatile basic nitrogen (VBN) levels, bacterial counts and electrophoretic behavior. Changes in pH, bacterial counts, VBN and K-values were associated with increased storage temperature and time. The levels of pH values, bacterial counts, VBN and K-values of pork samples stored at $25^{\circ}C$ were higher than those of the pork samples stored at $-2^{\circ}C$. ATP concentration decreased faster in samples stored at $25^{\circ}C$. Only IMP increased in samples stored at $-2^{\circ}C$, whereas the concentration of hypoxanthine and inosine increased in samples stored at $25^{\circ}C$. One exothermic peak and two endothermic peaks appeared on the thermograms of pork stored at either temperature. Lower transition temperature of myosin, sarcoplasmic protein and actin peaks were observed. The freshness parameters of K-value, VBN and hypoxanthine showed highly negative correlations (-0.742- -0.9980) to the changes in transition temperature. Therefore, the shift temperature on DSC thermogram can be used as an indicator of the freshness parameters of meat.

• 한국환경농학회지
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• 제32권1호
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• pp.48-54
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• 2013

수수 종자의 품종 간 특이적으로 발현하는 단백질을 동정하여 기능성 유전자를 확보하고 이들 유전자를 이용하여 수수의 기능성 강화 및 품종 판별 기술 개발을 위한 유용 유전자를 확보하고자 프로테오믹스 기법을 이용하여 수수 종자로부터 단백질을 추출하였다. 추출한 단백질을 이차원전기영동후, colloidal CBB 염색을 통해 품종 별로 발현에 차이를 보이는 단백질을 분석하였다. 총 652 개의 spot들 중에 8개의 단백질 spot들이 발현 정도에 변화를 보였으며, 이들 단백질을 MALDI-TOF/TOF MS와 MASCOT database를 통해 동정한 결과, RNA metabolism (spot 1, spot 4) HSP (spot 2), 저장 단백질 (spot 3, spot 5, spot 6), 산화-환원 (spot 8) 관련 단백질 등이 동정되었다. 특히 동정된 단백질은 주로 흰찰수수 (WCS)에서 발현 정도가 높게 나오는 경향을 보였으며, 흰찰수수 (WCS)에서 유일하게 발현 되는 단백질로 Cupin family protein, Gloubulin 등이 동정되었다. DEAD-box helicase는 흰찰수수 (WCS)를 제외한 나머지 세 품종에서 발현되었다. Ribonuclease T2와 Aldo-Keto reductase는 대풍수수 (DPS)를 제외한 나머지 세 품종에서 발현되었다. HSPs는 토종수수 (TJS)에서만 발현 되는 것을 확인하였다. 이들 동정된 단백질들은 수수의 품종 별 특성을 이해하는데 중요한 단서를 제공할 것으로 예측된다.