• 한국작물학회지
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• 제60권1호
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• pp.41-46
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• 2015

본 연구는 국내 육성 품종인 태광콩의 등숙기에 따른 단백질 발현 양상을 비교함으로써 등숙기 단백질 발현의 차이에 대한 기초자료를 얻고자 수행하였다. 동한 개화 후 종실의 등숙이 진행됨에 따라서 단백질 발현 양상이 세가지 경향으로 나뉘어 지는 것을 확인하였다. 첫 번째는 등숙이 진행됨에 따라서 단백질 발현 정도가 증가하다가 감소되며, 두 번째는 증가와 감소의 시기가 성숙기에 이루어지며, 세번째는 등숙기부터 성숙기까지 점진적으로 증가하는 것이다. 이러한 현상은 단백질의 기능에 따라 달라지는 것으로 사료된다. 등숙 초기에는 등숙에 필요한 단백질의 발현이 증가할 것이며 등숙 후기에는 저장단백질의 발현이 증가할 것으로 사료된다. 따라서 향후 좀 더 많은 수의 단백질 spot 들을 동정하여 어떤 기능을 가진 단백질이 등숙기에 따라 단백질의 발현 양상이 달라지는지는 좀 더 면밀히 관찰할 필요성이 있다고 사료된다.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2017년도 9th Asian Crop Science Association conference
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• pp.86-86
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• 2017

Prolamins are the main seed storage proteins in cereals. Gluten proteins seem to be prolamins because their primary structure have the meaningful quantity of proline and glutamine amino acid residues. Gluten proteins are found in crops such as wheat (Triticum aestivum), barley (Hordeum vulgare), and rye (Secale cereale) which are major food crops in the Triticeae tribe. Glutenin and gliadin, hordein, and secalin are typical gluten proteins found in wheat, barley, and rye, respectively. Gluten affect grain quality so that many researches, such as isolation or characterization of their genes, have been carried out. To improve the quality of grains in the Triticeae tribe, it is necessary to understand the relationship within their gluten proteins and their evolutionary changes. The sequences of nucleotides and amino acids of gluten protein including glutenins, gliadins, hordeins, and secalins were retrieved from NCBI (https://www.ncbi.nlm.nih.gov/) and Uniprot (http://www.uniprot.org/). The sequence analysis and the phylogenetic analysis of gluten proteins were performed with various website tools. The results demonstrated that gluten proteins were grouped with their homology and were mostly corresponded with the previous reports. However, some genes were moved, duplicated, or disappeared as evolutionary process. The obtained data will encourage the breeding programs of wheat, barley, rye, and other crops in the Triticeae tribe.

• 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.

• Journal of Plant Biotechnology
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• 제38권4호
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• pp.263-271
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• 2011

벼는 세계 인구의 60%에 의해 소비되고 있는 주요 식량작물이며 그 종자의 주성분은 탄수화물로 인류의 중요한 에너지원이 된다. 미곡(米穀)은 주식으로 다량 섭취하게 되는데 특히 동물성 단백질의 섭취가 부족한 국가 또는 지역에서는 쌀 단백질이 콩 단백질과 함께 중요한 영양공급원이 되고 있어 벼의 종자단백질은 인류에 매우 중요한 영양성분이라 할 수 있다. 그런데 벼의 종자단백질은 필수아미노산인 라이신이 부족하므로 아미노산 조성 변경에 의한 영양적인 개량이 요구되기도 하는 한편 선진국에서는 혈압조절이나 면역증강 등 생리기능을 가진 건강증진용 기능성 단백질 또는 펩티드로 주목받고 있다. 따라서 벼의 종자단백질의 조성변경과 더불어 이종의 저장단백질의 도입에 의한 벼 종자단백질 개량 연구가 진행되어 왔다. 본 총설에서는 벼의 종자 저장단백질의 생합성과 축적 특징 및 저장단백질 집적의 유전적 제어 기작에 대하여 알아보고 또한 벼 종자 저장단백질 조성 변경, 이종단백질 도입에 의한 벼 종자 저장단백질 개량 연구 현황을 기술하고자 한다.

• 생명과학회지
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• 제8권3호
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• pp.318-325
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• 1998

To investigate changes in protein and total lipid contents, seed storage protein pattern, and fatty acid composition of germination perilla(Perilla frutescens) seeds. Also, the corresponding value components in cotyledons, hypocotyles and roots were measured according to germination stage. The results were summarized as follows ; During germination, pertein and total lipid contents of Yepsilldalggae and Kwangyang cultivar were decreased continuously. In particular, protein contents rapidly decreased to the 3 days after germination(DAG), and then total lipid contents rapidly decreased between 3 DAG and 10 DAG. In changes of protein and total lipid contents of cotyledons, protein contents of Yeupsildalggae was increased during the germination, but Kwangyang cultivar was decreased during the same periods. The total lipids contents of Yeupsildalggae and Kwangyang cultivar were decreased during the germination. According to SDS-PAGE analysis, there was no detectible polypeptide bands on the gel before seed germination suggesting that this may be due to the rapid degradation of the storage proteins in the mature seed by hydrolyttic enzymes during the stage. During germinatation , the polypeptide band with 27$\sim$28KD of Yeupsildalggae and Kwangyang cultivar were accumulated gradually. In changes of fatty acid composition of total lipid of Yeupsildalggae and Kwangyang cultivar , saturated fatty acids such as palmitic acid and stearic acid increased during the germination. On the other hand, unsaturated fatty acid such as linoleic acid and linolenic acid decreased during the same periods. However, oleic acid increased to the 5 DAG, and then was repidly decreased.

• 식물조직배양학회지
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• 제24권4호
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• pp.197-201
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• 1997

Plant seed oil-bodies or oleosomes ate the repository of the neutral lipid stored in seeds. These organelles in many oilseeds may comprise half of the total cellular volume. Oleosomes are surrounded by a half-unit membrane of phospholipid into which are embedded proteins called oleosins. Oleosins are present at high density on the oil-body surface and after storage proteins comprise the most abundant proteins in oilseeds. Oleosins are specifically targeted and anchored to oil-bodies after co-translation on the ER. It has been shown that the amino-acid sequences responsible for this unique targeting reside primarily in the central hydrophobic tore of the oleosin polypeptide. In addition, a signal-like sequence is found near the junction of the hydrophobic domain and ann N-terminal hydrophilic / amphipathic domain. This "signal" which is uncleaved is also essential for correct targeting. Oil-bodies and their associated oleosins may be recovered by floatation centrifugation of aqueous seed extracts. This simple partitioning step results in a dramatic enrichment for oleosins in the oil-body fraction. In the light of these properties, we reasoned that it would be feasible to create fusion proteins on oil-bodies comprising oleosins and an additional valuable protein of pharmaceutical or industrial interest. It was further postulated that if these proteins were displayed on the outer surface of oil-bodies, it would be possible to release them from the purified oil-bodies using chemical or proteolytic cleavage. This could result in a simple means of recovering high-value protein from seeds at a significant (i.e. commercial) scale. This procedure has been successfully reduced to practice for a wide variety of proteins of therapeutic, industrial and food no. The utillity of the method will be discussed using a blood anticoagulant, hirudin, and industrial enzymes as key examples.

• 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.

• 한국식물학회:학술대회논문집
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• 한국식물학회 1987년도 식물생명공학 심포지움 논문집 Proceedings of Symposia on Plant Biotechnology
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• pp.261-282
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• 1987

Plants store a significant amount of their nitrogen, sulfur and carbon reserves as storage proteins in seed tissues. The major proteins present in rice seeds are the glutelins. Glutelins are initially synthesized at 4-6 days postanthesis and deposited into protein bodies via Golgi apparatus. Based on nucleic acid sequences and Southern blot analysis, the three isolated glutelin genomic clones were representative members of three gene subfamilies each containing 5 to 8 copies. A comparison of DNA sequences displayed by relevant regions of these genomic clones showed that two subfamilies, represented by clones, Gt1 and Gt2, were closely, related and probably evolved by more recent gene duplication events. The 5' flanking and coding sequences of Gt1 and Gt2 displayed at least 87% homolgy. In contrast, Gt3 showed little or no homolgy in the 5' flanking sequences upstream of the putative CAAT boxes and exhibited significant divergence in all other portions of the gene. Conserved sequences in the 5' flanking regions of these genes were identified and discussed in light of their potential regulatory role. The derived primary sequences of all three glutelin genomic clones showed significant homology to the legume 11S storage proteins indicating a common gene origin. A comparison of the derived glutelin primary sequences showed that mutations were clustered in three peptide regions. One peptide region corresponded to the highly rautable hypervariable region of legume peptide region of legume 11S storage proteins, a potential target area for protein modification. Expression studies indicated that glutelin mRNA transcripts are differentially accumulated during endosperm development. Promoterss of Gt2 and Gt3 were functional as they direct transient expression of chloramphenicol acetyltransferase in cultured plant cell.

• 한국작물학회지
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• 제61권4호
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• pp.227-234
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• 2016

본 연구에서는 쌀의 저장 단백질 조성 분석을 통해 자포니카와 통일형 품종의 식미 특성 차이를 구명하고자 하였다. 쌀의 저장단백질 분석 결과, 통일형 품종은 자포니카 품종에 비해 알부민과 글로부린이 각 1.2배, 1.3배, 글루텔린의 서브유닛인 산성 글루텔린 ${\alpha}-2$의 비율이 약 1.3배 낮은 반면, 글루텔린의 서브유닛인 산성 글루텔린 ${\alpha}-1$ 비율은 1.7배 높은 특성을 나타냈다. 또한, 저장단백질 조성과 식미특성의 상관성 분석을 실시한 결과, 식미 총평은 알부민(R=0.495, p<0.01), 글로부린(R=0.567, p<0.01)과 정의상관을 보였으며 산성 글루텔린 유닛인 ${\alpha}-1$은 총평과 뚜렷한 부의 상관(R=-0.612, p<0.01)이 인정되었다. 식미 총평 이외에도 밥의 외관, 맛, 찰기, 질감과 관련하여 알부민은 정의 상관, ${\alpha}-1$ 글루텔린은 부의 상관관계를 나타냈다. 생태형에 따른 저장단백질 비율이 식미 특성에 미치는 영향을 구명하기 위해 저장단백질 비율과 식미 특성과의 상관 분석을 실시한 결과, 산성 글루텔린의 서브유닛인 ${\alpha}-1$, 알부민, 글로부린이 통일형 품종의 식미 특성에 영향을 주는 것으로 보이며 향후, 다양한 품종을 이용한 추가 연구를 통해 정밀한 해석이 필요할 것으로 생각된다.

• 한국작물학회지
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• 제47권2호
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• pp.85-94
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• 2002

Antibodies raised against the purified p-subunit of $\beta$-conglycinin were used in immunohistochemical studies to monitor the pattern of $\beta$-conglycinin mobilization in the cotyledons during soybean [Glycine max (L.) Merr.] seed germination. Western blot analysis revealed that the break down of the $\beta$-subunit of $\beta$-conglycinin commenced as early as 2 days after seed imbibition (DAI). Concurrent with the degradation of the $\beta$-subunit of $\beta$-conglycinin, accumulation of 48, 28, and 26 kD proteolytic intermediates was observed from 2 to 6 DAI. Western blot analysis also revealed that the acidic subunit of glycinin was mobilized earlier than the basic subunit. The basic glycinin subunit was subjected to proteolysis within 2 DAI resulting in the appearance of an intermediate product approximately 2 kD smaller than the native basic glycinin subunit. In contrast to the major seed storage proteins, lipoxygenase was subjected to limited proteolysis and was detected even after 8 DAI. The first sign of $\beta$-conglycinin breakdown was observed near the vascular strands and proceeded from the vascular strands towards the epidermis. Protein A-gold localization studies using thin sections of soybean cotyledons and antibodies raised against the $\beta$-subunit of $\beta$-conglycinin revealed intense labeling over protein bodies. A pronounced decrease in the protein A-gold labeling intensity over protein bodies was observed at later stages of seed germination. The protein bodies, which were converted into a large central vacuole by 8 DAI, contained very little 7S protein as evidenced by sparse protein A-gold labeling in the vacuoles.