• Title/Summary/Keyword: 43 kD protein

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Biochemical Studies on the Sugar Chain Structure of Glycoproteins with the Same Protein Core of Bovine Milk Fat Globule Membrane (공통의 1차 구조를 가진 우유 지방구막 구성단백질의 당쇄 구조에 관한 생화학적 연구)

  • Seok, Jin-Seok
    • Journal of Dairy Science and Biotechnology
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    • v.21 no.2
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    • pp.138-147
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    • 2003
  • We here analyzed and proposed the structures of the N-linked sugar chains of PAS-7 from bovine milk fat globule membrane. The N-linked sugar chains were liberated from PAS-7 by hydrazinolysis and, after modifying the reducing ends with 2-aminopyridine (PA), were separated into one neutral (7N,55%) and two acidic (7M mono-, 43%; 7D, di-, 2%) sugar chain roups. The latter were converted into neutral groups (7MN and 7DN) by sialidase digestion. The structure of each of these PA-neutral sugar chains was determined by sugar analysis, sequential exoglycosidase digestion, partial acetolysis, and 1H-NMR spectroscopy. The results show that the 10 sugar chains were of the biantennary complex type with and without fucose. The structure of 7N2A one of the major sugar chains, was proposed as; [structure: see text] A structural comparison between PAS-6 and -7 indicated that although they shared the same protein core, their sugar moiety was markedly different, involving the existence of a different pathway during the post-transcriptional modification.

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Production of monoclonal antibodies against VP28 of white spot syndrome virus (WSSV) (White spot syndrome virus (WSSV)의 VP28에 대한 단클론 항체 생산)

  • Bang, Ji-hyeong;Kim, Wi-Sik;Kim, Choon-sup;Kim, Jong-Oh;Oh, Myung-Joo
    • Journal of fish pathology
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    • v.32 no.1
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    • pp.45-48
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    • 2019
  • We developed and subsequently characterized mouse monoclonal antibodies (MAbs) against recombinant VP28 structural protein (rVP28) of white spot syndrome virus (WSSV). We established six hybridoma clones secreting MAbs against rVP28: 15A11, 20G6, 31H2, 34H6, 38D1 and 43A1. All six MAbs recognized the 25 kDa of protein in gill homogenates of WSSV-infected shrimp by western blot analysis, while no reactivity was observed in gill homogenates of normal shrimp. Moreover, high enzyme-linked immunosorbent assay (ELISA) optical density (OD) values (0.8-2.68) were observed in the hemolymphs from WSSV-infected shrimp, while low OD values (less than 0.24) were recorded in the hemolymphs from normal shrimp, by using these six MAbs produced in this study. These results suggest that these six MAbs are useful for the detection of WSSV.

Diversity in Lipid Contents and Fatty Acid Composition of Soybean Seeds Cultivated in Korea (국내 유통 콩의 지방함량 및 지방산 조성변이)

  • Kim, Sun-Lim;Lee, Yeong-Ho;Chi, Hee-Youn;Lee, Sun-Joo;Kim, Si-Ju
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.52 no.3
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    • pp.348-357
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    • 2007
  • The 117 soybeans seeds were collected from the nine provinces of Korea, and protein and lipid contents, and fatty acid composition levels were evaluated to investigate their relationship. The 100-seed weights of the black soybeans were varied $27.7{\sim}33.1g$, while the 100-seed weight of yellow soybeans were varied $24.6{\sim}36.6g$. Protein and lipid content of the 117 soybean seeds was 38.3% and 17.8%, respectively Protein contents of the 59 black soybean seeds (38.6%) were significantly higher than those of the 58 yellow soybean seeds (37.9%). However, lipid contents of the black soybean seeds (17.6%) were lower than those of the yellow soybean seeds (18.1%). Linoleic and oleic acid composition levels of the 117 soybean seeds were 53.75% and 22.08%. Unsaturated fatty acid levels of soybean seeds showed a statistically significant variability among the nine provinces of Korea, however, the differences were not found in the linoleic (18:2) and oleic acid (18:1) levels. Therefore, it was considered that the significant variability of unsaturated fatty acid were mainly due to the variations of linolenic acid (18:3) level. The composition levels of linoleic, oleic, palmitic (16:0), and linolenic acid in the yellow soybean seeds were 53.43%, 22.73%, 12.23%, and 8.24%, while those of the black soybean seeds were 54.13%, 21.48%, 12.47%, and 8.31%. Obtained results suggested that fatty acid composition levels were varied and possibly influence by the phenotype of seed coat colors. Oleic acid, mono-saturated fatty acid, showed the most remarkable variability between yellow and black soybean seeds, and the composition levels were higher in the yellow soybean seeds. Relationship between unsaturated fatty acid levels and 100-seed weights in the yellow soybean seeds showed a negative correlation (r=-0.513, P<0.01), but no relationship $(r=0.154^{ns})$ was observed in the black soybean seeds.

Electrophoretic and Immunological Evaluation of Secalin in Rye, Triticale, and Wheat-Rye Translocation Wheat

  • Seo, Yong-Weon;Hong, Byung-Hee
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.43 no.4
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    • pp.228-233
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    • 1998
  • Seed storage proteins have been used for studying biochemical genetics and end-use quality aspects. We conducted enzyme-linked immunosorbent assay (ELISA) and one-dimensional SDS-PAGE (1D SDS-PAGE) to evaluate different cereal crop species and Korean wheat lines for rye secalin proteins. The antisecalin antibody showed consistent specificity for rye secalin with little cross-reactivity to gliadins. Immunological cross-reactivities measured by the ELISA technique using competition assay showed significant differences of absorbance among rye, triticale, wheat-rye translocated wheat and non-translocated wheat. The absorbance values were lowest in rye followed by triticale, translocated wheat and non-translocated wheat. The ELISA for discrimination of wheat-rye translocation on the basis of antigen-antibody reactivity showed that none of the Korean wheat lines possessed 1RS and secalin proteins. The competitive ELISA experiment demonstrated specific determination for secalin that was originated from rye chromosomal parts. The result of 1D SDS-PAGE for identifying rye secalin subunits showed all three rye specific secalin protein subunits (75 KDa, 45 KDa, and 40 KDa) for rye and triticale, and 1RS specific secalins (45 KDa and 40 KDa) for 1AL/1RS and 1BL/1RS translocated wheats. All Korean wheats were lacking 1RS of rye chromosome and secalin.

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한우 난자의 체외성숙 시간에 따른 세포질 내 단백질 합성의 변화

  • 박용수;박흠대;변명대
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.32-32
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    • 2003
  • 소 난자의 체외성숙 과정에서 세포질 내 단백질의 생산과 축적의 변화는 핵 및 세포질 성숙과 밀접한 관계가 있다는 보고가 있지만, 난자의 성숙과 관련된 특정 단백질의 종류에 대한 보고는 없었다. 따라서 본 연구는 한우 난자의 체외성숙과 관련된 단백질의 생산 및 축적의 변화와 그 종류에 대해서 검토하였다. 체외성숙 시간(4.5, 9, 13.5, 18 및 24시간)에 따른 배양액 내의 단백질 합성의 변화는 2D gel electrophoresis를 이용하였고, 단백질 spot에 대해서는 peptide mass fingerprinting(PMF) 방법을 이용하였다. 또한 단백질 측정 시간에 신선 체외성숙 배양액으로 교환 후 난포란의 핵성숙과 배발달율을 검토하였다. 그 결과 한우 난포란의 체외성숙 시간에 따라 배양액에서 단백질의 양 및 질적인 변화를 확인하였다. 그리고 총 296개 단백질 spot들을 확인하였고, 그 중 30개 spot에서 유의적인 변화가 인정되었다. 또한 유의적인 변화를 보인 spot에 대한 PMF 분석을 통하여 Apolipoprotein A-1 precursor, Alpha enolase, Aldose reductase, 43kDa collectin precursor, Heat shock 27kDa protein, Plasminogen activator inhibitor-1 precursor, Thrombospondin 1 및 Transitional endoplasmic reticulum ATPase가 동정되었다 그리고 총 단백질 합성 경향은 0∼4.5 시간에는 감소하였고, 13.5∼18시간에 증가 한 후 다시 감소하는 경향을 나타내었으며, 단백질의 종류도 시간대별로 현저한 변화가 있었다. 한편 단백질을 측정하는 시기에 신선 체외성숙 배양액으로 교환한 후 난포란의 핵성숙 및 배발달율을 검토한 결과 18시간 체외성숙군에서 9시간째의 교환이 유의적으로 높은 핵성숙을 나타내었으나, 배발달율에서는 유의성이 인정되지 않았다. 그러나 24시간 체외성숙군에서 18시간째의 배양액 교환은 8세포기 및 배반포 발달율이 유의적(P<0.05)으로 높았다. 연구 결과로부터 소 난자의 체외성숙 시간에 따른 단백질 합성 경향의 차이를 확인하였고, 유의적인 변화를 나타낸 8가지의 단백질을 분리할 수 있었으며, 향후 이들 작용기전에 대한 연구가 필요하다고 사료된다.

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Mosquito Larvicidal Activity of Synechocystis PCC6803 Transformed with the cry11Aa gene to Culex tritaeniorhynchus and Anopheles sinensis (Cry11Aa 유전자로 형질전환된 Synechocystis PCC6803의 작은빨간집모기와 중국얼룩날개모기 유충에 대한 살충효과)

  • 이대원
    • Korean journal of applied entomology
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    • v.43 no.1
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    • pp.35-41
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    • 2004
  • Bacillus thuringiensis produces crystal proteins toxic to medically and agriculturally important pests during sporulation. To improve the activity of insecticidal crystal protein in applying to mosquito larval control, an expression vector, pSyn4D harboring the mosquitocidal cry11Aa gene under control of psbA promoter of Amaranthus hybridus was constructed. This expression vector was transformed into Synechocystis PCC6803 and a transformant, Tr2C was selected with kanamycin. The mosquitocidal cry11Aa gene was stably integrated Into genomic DNA of Tr2C in PCR detection using cry11Aa-specific primers. The transformant expressed 72-kDa Cry11Aa protein and median lethal time (LT$\sub$50/) was approximately 2.1 days for Culex tritaeniorhynchus larvae and 0.7 day for Anopheles sinensis larvae, respectively. These results suggest this transformant can be used for mosquito larval control as a biological control agent.

Pheontypic Characterization of Non-molting Mutants in Postembryonic Development of the Silkworm, Bomyx mori. (누에의 후배자발생 초기에 있어서 돌연변이 불면잠계통간의 형질발현 특성)

  • No, Si-Gap;Seon, Hui-Suk;Banno, Yutaka
    • Journal of Sericultural and Entomological Science
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    • v.40 no.1
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    • pp.43-51
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    • 1998
  • Studies were carried out to investigate phenotypic expression, mortality and biochemical analysis of haemolymph proteins of nm-d, nm-f, nm-k and nmn bib-molting mutants of the silkworm, Bombyx mori. The non-molting mutants characters were expressed in the homozygote of each mutant genes. All strains of non-molting mutants were similar with each other in physiological characteristics, but the expression varied with each strains. The larvae of nm-d, nm-i and nmn died between day 5 and day 9 after hatching without the first molt. The nm-f and nm-k mutants died between day 5 and day 16 with a slight increase of body weight and, more than 90% of the mutants larvae died before the first molt and a few of them survived to the 2nd and the 3rd instar and died. The haemolymph protein components of nm-d, nm-i, and nmn were rapidly reduced, and on the other hand those of nm-f and nm-k consistently until they died. And there were no distinguishable difference in haemolymph components of non-molting mutants, as compared to those normals.

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Processing and Fermentation of Food Wastes with MS Microorganism Complex for Swine Feeds (MS 미생물복합군에 의한 음식폐기물의 양돈사료화를 위한 적정처리공정)

  • Lee, Jeong-Chae;Jung, Woo-Jin;Lim, Kye-Taek;Kim, Tae-Hwan
    • Korean Journal of Environmental Agriculture
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    • v.19 no.4
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    • pp.270-275
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    • 2000
  • In order to investigate the proper processing of food wastes with miraculous soil-microorganisms (MS) for final use of swine feeds, calory, amino acid and fatty acid in food wastes were determined in relation with fermentation process with MS microorganism complex. Aflatoxin test was also performed to check safety of the fermented food wastes. Calory of food wastes was determined in average $7.60\;Kcal{\cdot}g^{-1}\;D.W.$ In finally processed food wastes, total content of amino acid was $93.0\;mg{\cdot}g^{-1}]\;D.W$, showing 18.5% of increase by the anaerobic fermentation. Essential and non-essential amino acids were measured at respectively 34.43 and $58.56\;mg{\cdot}g^{-1}\;D.W.$ Leucine, phenylalanine, isoleucine and threonine of essential amino acids and proline and glutamic acid of non-essential amino acids were highly composed as compared to others. The composition of fatty acid in food wastes was also increased by anaerobic fermentation for 3 weeks. Palmitic acid, oleic acid and palmitoleic acid were more important in quantity. Present results indicate that food wastes properly processed with MS have enough calory and are safe from aflatoxin, and that anaerobic fermentation with MS microorganism in an efficient process for hydrolyzing protein and lipids in food wastes.

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Indentification of Specific Proteins synthesized During Somatic Embryogenesis of Rice (벼 callus로부터 재분화 과정에서 생성되는 특수한 단백질의 동정)

  • Won, Jae Hee;Choi, Hong Jib;Kim, Dal Ung
    • Current Research on Agriculture and Life Sciences
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    • v.9
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    • pp.95-102
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    • 1991
  • This study was conducted to investigate the physiological and biochemical elements relating embryogenisis. Also we examined the differences of protein in embryogenic callus and nonembryogenic callus by SDS-PAGE and two-dimensional gel electrophoresis. In this study, we have focused attention on the process of total proteins during sometic embryogenesis of rice. When compared the proteins of embryogenic callus and nonembryogenic callus, some different proteins were observed and serval proteins were increased in embryogenic callus. Some proteins were decreased or disappeared in embryogenic callus. Near 43kD protein band was only observed in embryogenic callus. The other bands were similar to each lines. The result of two-diimensional gel electrophoresis, E-callus specific proteins were observed. This Results may indicate that these proteins were associated with somatic embryogenesis.

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Synthesis, Characterization and Cosmetic Application of Self-Assembled Sericin-PEG Nanoparticle

  • E. S. Choung;S. Y. Eom;Kim, J. H.;Kim, K. S.;Kim, K. H.;Lee, K. G.;Lee, Y. W.;C. S. Cho
    • Proceedings of the SCSK Conference
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    • 2003.09a
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    • pp.501-519
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    • 2003
  • Silk Sericin(SS) is a natural protein extracted from cocoon of bombix mori and shows moisturizing effect to the skin due to a number of hydroxyl groups in the structure. But its application to cosmetics is limited due to its poor solubility in water. In order to solve this drawback and expand its application to cosmetics, polyethyleneglycol(PEG) was conjugated with sericin by reacting activated polyethyleneglycol(ActPEG). Reaction site of sericin is tyrosine residue, which was determined by using $^1$H-NMR. Random coil structure of sericin was transformed to beta-sheet structure by conjugating polyethyleneglycol. It was confirmed that melting point of sericin-PEG conjugate was lowered compared to that of each sericin and PEG due to the interaction between sericin and PEG in crystalline structure. Self-assembled sericin-PEG nanoparticle was obtained by dialyzing with alcohol solution of sericin-PEG conjugate against water. The particle is spherical and has 200-400nm of size. The moisturizing ability of sericin-PEG nanoparticle was much higher than that of sericin itself. Incorporation of vitamin A into sericin-PEG nanoparticle was carried out by diafiltration method. The content of incorporated Vitamin A in sericin-PEG nanoparticle was 8.9 wt%. Releasing behaviour of vitamin A incorporated into nanoparticle was tested in phosphate buffer, pH 7.4 at 37$^{\circ}C$. and half-life of Vitamin A release was 43hrs. Sericin-PEG nanoparticle exhibited higher moisturing effect than sericin itself and distilled water, respectively. No toxicity and irritation were observed in animal tests. It can be expected that the self-assembled sericin-PEG nanoparticle can be developed for cosmetics.

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