• Title/Summary/Keyword: 30K protein

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Separation and Purification of Lysozyme from Chicken Eggwhite Through Ultrafiltration (한외여과를 통한 난백 중 라이소자임의 분리정제)

  • Koo Ja-Kyung;Son Dongho;Jun Hoejin;Lee Yunhee;Cho Namjun;Jang Dong Il
    • Membrane Journal
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    • v.15 no.2
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    • pp.121-131
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    • 2005
  • Separation and purification of lysozyme from chicken egg white was studied using ultrafiltration. We have obtained experimental data through the cellulose membranes with the molecular weight cut off (MWCO) of 10 kDa, 30 kDa and 100 kDa in a stirred ultrafiltration device. Certain amounts of egg white were dissolved into 20 mM phosphate buffers of pH 6, 7 and 8 to make protein solutions of $1\%,\;2\%,\;3\%\;and\;10\%$ concentration. Permeation flux increased with increasing MWCO of the membrane. Permeation flux increased with increasing transmembrane pressure (TMP) and decreasing the protein concentration. As the MWCO of membrane decreased, the selectivity increased. The selectivity increased with increasing TMP and protein concentration of the solution.

Antigenic Proteins of Helicobacter pylori of Potential Diagnostic Value

  • Khalilpour, Akbar;Santhanam, Amutha;Lee, Chun Wei;Saadatnia, Geita;Velusamy, Nagarajan;Osman, Sabariah;Mohamad, Ahmad Munir;Noordin, Rahmah
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.3
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    • pp.1635-1642
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    • 2013
  • Helicobacter pylori antigen was prepared from an isolate from a patient with a duodenal ulcer. Serum samples were obtained from culture-positive H. pylori infected patients with duodenal ulcers, gastric ulcers and gastritis (n=30). As controls, three kinds of sera without detectable H. pylori IgG antibodies were used: 30 from healthy individuals without history of gastric disorders, 30 from patients who were seen in the endoscopy clinic but were H. pylori culture negative and 30 from people with other diseases. OFF-GEL electrophoresis, SDS-PAGE and Western blots of individual serum samples were used to identify protein bands with good sensitivity and specificity when probed with the above sera and HRP-conjugated anti-human IgG. Four H. pylori protein bands showed good (${\geq}$ 70%) sensitivity and high specificity (98-100%) towards anti-Helicobacter IgG antibody in culture-positive patients sera and control sera, respectively. The identities of the antigenic proteins were elucidated by mass spectrometry. The relative molecular weights and the identities of the proteins, based on MALDI TOF/TOF, were as follows: CagI (25 kDa), urease G accessory protein (25 kDa), UreB (63 kDa) and proline/pyrroline-5-carboxylate dehydrogenase (118 KDa). These identified proteins, singly and/or in combinations, may be useful for diagnosis of H. pylori infection in patients.

Involvement of Phosphatidylinositol 3-Kinase in the Insulin Signaling in Preimplantation Mouse Embryos (생쥐 착상전 배아의 인슐린 신호전달 과정에 Phosphatidylinositol 3-Kinase의 관련성)

  • Gye, Myung-Chan;Nah, Hee-Young;Kim, Moon-Kyoo
    • Development and Reproduction
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    • v.4 no.1
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    • pp.29-35
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    • 2000
  • A phosphatidylinositol 3-kinase (PI3K) is a upstream component of insulin signaling by which protein synthesis can be stimulated in many systems. To elucidate involvement of PI3K and its downstream mammalian target of rapamycin (mTOR) in the insulin signaling in pleimplantation mouse embryos, 8-cell embryos were cultured to blastocysts in the presence or absence of insulin and/or inhibitor drugs. The number of blastomeres per blastocyst, protein synthesis, and protein phosphorylation were examined. There was significant difference in embryonic development to blastocyst stage and hatching was potentiated by the insulin supplementation. The increase in the mean celt numbers per blastocyst was apparent in the insulin culture. Wortmannin, a PI3K inhibitor and rapamycin, an inhibitor of mTOR abolished the stimulatory effect of insulin on morphological development mitosis and protein synthesis. In autoradiography, phosphoproteins pp22 and pp30 which undergo phosphorylation in response to insulin were identified. Taken together, it can be suggested that PI3K and mTOR engaged in insulin signaling in the mouse embryo 8-cell onward and mediate embryotropic offset of insulin.

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Expression of a Fusion Protein with Cry1Ac Protein and a Scorpion Insect Toxin in Acrystalliferous Bacillus thuringiensis Strain

  • Roh, Jong-Yul;Li, Ming-Shun;Chang, Jin-Hee;Park, Jae-Young;Shim, Hee-Jin;Shin, Sang-Chul;Boo, Kyung-Saeng;Je, Yeon-Ho
    • International Journal of Industrial Entomology and Biomaterials
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    • v.8 no.1
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    • pp.89-93
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    • 2004
  • Expression of a fusion protein between B. thuringiensis crystal protein, Cry1Ac1 and a scorpion insect toxin (AaIT, Androctonus australis Hector insect toxin) in acrystalliferous B. thuringiensis strain (Cry-B strain) was examined. The cry 1Ac1 gene was cloned in B. thuringiensis-E coli shuttle vector, pHT3101, under the control of the native cry 1Ac1 gene promoter (pProAc) and a gene encoding AaIT was inserted in XhoI site in the middle of the cry 1Ac1 gene (pProAc-ScoR). B. thuringiensis Cry-B strain carrying pProAc-ScoR (PyoAc-ScoR/CB) produced an inclusion body of irregular shape and the expressed fusion protein is approximately 65 kDa in size. Sporulated cells and spore-crystal mixtures of ProAc-ScoR/CB had insecticidal activity against Plutella xylostella larvae, showing $LT_50$ of ProAc-ScoR/CB (22.59 hrs) lower than that of ProAc/CB (30.06 hrs) at $1{\times}{10^7} {CEU/cm^2}$. These results suggest that the fusion protein including a B. thuringiensis crystal protein and an AaIT may be functionally expressed in B. thupingiensis. Moreover, we verified the additive toxicity of AaIT, which is a new feasible candidate for insect control.

Fermentation and Purification of LacZ-Fused Single Chain Insulin Precursor for($B^{30}$-Homoserine) Human Insulin

  • SeungYup Lee;Jeo
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.1 no.1
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    • pp.9-12
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    • 1996
  • In order to produce the single chain precursor of a novel human insulin analogue, (B30-Homoserine) insulin, the fermentative behaviors of Escherichia coli JM103 were studied, which harbors pKBA plasmid carrying a hybrid gene in which the gene for a single chain precursor was fused with lacZ gene under tac promoter. The maximal induction of gene expression was achieved when more than 0.05 mM of isopropyl-$\beta$-D-thiogalactopyranoside(IPTG) was supplemented to fermentation medium after 4 h cultivation of E. coli, and followed by longer than 2-h fermentation. The hybrid protein of the single chain insulin precursor was isolated from cytoplasmic inclusion bodies by dissolving in 8M urea solution, and purified through DEAE-Sephacel and Sephadex G-200 column chromatographies with a recovery of 35%. The finally purified hybrid protein showed a single band on sodium dodecyl sulfate-polyacrylamide gel.

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Biological Control of Plant Diseases With Bacillus Species

  • Li, Debao
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 1994.06a
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    • pp.75-85
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    • 1994
  • Biocontrol is playing a more and more important role in plant disease management. Evidences show that there are optimum prospects for people to apply biocontrol approach to control plant disease or to study the mechanism of antagonism.“The study of Antagonistic Protein of Bacillus spp.to Xanthomonas oryzae pv. oryzae”has been worked in our laboratory since 1986. One hundred and thirty antagonistics bacteria were screened out, most of them belonged to Bacillus spp., and showed very strong inhibitive effect to various plant pathogens. Nine antagonistic proteins (peptides) were purified (P11-I, P11-II, B8, B826-I, B826-II, A30-I, A30-II, G35). Two antagonistic protein related DNA fragments (B826-I, A30-II) were cloned and sequenced. B826-I DNA fragment composed by 905 bp, and it contained two ORF encoding 95, and 54 amino acids, respectively. By using Rifr and Kamr as the selective markers, we found the bacteria could colonize on rice leaf for at least 40 days. In greenhouse the antagonistic bacteria showed certain degree of control efficiency.

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The Role of Protein Kinases in Reprogramming and Development of SCNT Embryos

  • Choi, Inchul;Campbell, Keith H.S.
    • Journal of Embryo Transfer
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    • v.30 no.1
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    • pp.33-43
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    • 2015
  • Successful somatic cell nuclear transfer (SCNT) has been reported across a range of species using a range of recipient cells including enucleated metaphase II (MII) arrested oocytes, enucleated activated MII oocytes, and mitotic zygotes. However, the frequency of development to term varies significantly, not only between different cytoplast recipients but also within what is thought to be a homogenous population of cytoplasts. One of the major differences between cytoplasts is the activities of the cell cycle regulated protein kinases, maturation promoting factor (MPF) and mitogen activated protein kinase (MAPK). Dependent upon their activity, exposure of the donor nucleus to these kinases can have both positive and negative effects on subsequent development. Co-ordination of cell cycle stage of the donor nucleus with the activities of MPF and MAPK in the cytoplast is essential to avoid DNA damage and maintain correct ploidy. However, recent information suggests that these kinases may also effect reprogramming of the somatic nucleus and preimplantation embryo development by other mechanisms. This article will summarise the differences between cytoplast recipients, their effects on development and discuss the potential role/s of MPF and or MAPK in nuclear reprogramming.

Investigating herbal active ingredients and systems-level mechanisms on the human cancers (암치료를 위한 네트워크 기반 접근방식 활용 시스템 수준 연구)

  • Lee, Won-Yung
    • Herbal Formula Science
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    • v.30 no.3
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    • pp.175-182
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    • 2022
  • Objective : This study aims to investigate the active ingredients and potential mechanisms of the beneficial herb on human cancers such as the liver by employing network pharmacology. Methods : Ingredients and their target information was obtained from various databases such as TM-MC, TTD, and Drugbank. Related protein for liver cancer was retrieved from the Comparative Toxicogenomics Database and literature. A hypergeometric test and gene set enrichment analysis were conducted to evaluate associations between protein targets of red ginseng (Panax ginseng C. A. Meyer) and liver cancer-related proteins and identify related signaling pathways, respectively. Network proximity was employed to identify active ingredients of red ginseng on liver cancer. Results : A compound-target network of red ginseng was constructed, which consisted of 363 edges between 53 ingredients and 121 protein targets. MAPK signaling pathway, PI3K-Akt signaling pathway, p53 signaling pathway, TGF-beta signaling pathway, and cell cycle pathway was significantly associated with protein targets of red ginseng. Network proximity results indicated that Ginsenoside Rg1, Acetic Acid, Ginsenoside Rh2, 20(R)-Ginsenoside Rg3, Notoginsenoside R1, Ginsenoside Rk1, 2-Methylfuran, Hexanal, Ginsenoside Rd, Ginsenoside Rh1 could be active ingredients of red ginseng against liver cancer. Conclusion : This study suggests that network-based approaches could be useful to explore potential mechanisms and active ingredients of red ginseng for liver cancer.

Selection Attributes and Purchasing Perceptions and Attitudes of Protein Snacks According to Individual Health Lifestyle (개인의 건강 라이프스타일에 따른 단백질 스낵의 선택속성과 구매인식 및 태도)

  • Hwang, Ji Eon;Oh, Ji Eun;Cho, Mi Sook
    • Journal of the Korean Society of Food Culture
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    • v.33 no.4
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    • pp.394-401
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    • 2018
  • This study was conducted to investigate the effects of health lifestyle on high protein snack selection attributes and purchase behaviors among individuals aged 20-30 with high protein snack intake. In addition, the relationship between perception, attitude, satisfaction and recommendation of high protein snacks was invested. Finally, this study aims to provide basic information for marketing high-protein snacks and customized high protein snacks. Analysis of the selection attributes most important for healthy lifestyle, revealed significant differences among all groups excluding the external seeking group (p<0.001). The free living group regarded trust as one of the most important attributes of high protein snack selection, and both the tempered control group and the low-interest group found sensation and price factor to be important. Therefore, when developing high-protein snacks, it is important to determine which attributes of the snack will be highlighted by segmenting the consumer into health lifestyles. Focusing on what ingredients are used to develop high-protein snacks and nutritional ingredients is also important when targeting a free lifestyle group as the main customer. In addition, developing snacks that do not offer depending on the protein content is important when targeting a temperate management group or a low-interest group.

Effect of Proteins Modified by Enzymically Oxidized Caffeic Acid on the Concentration of Serum Cholesterol of Rats (효소적 갈변 단백질이 흰쥐 혈청콜레스테롤 농도에 미치는 영향)

  • Cho, Young-Su
    • Applied Biological Chemistry
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    • v.37 no.5
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    • pp.379-384
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    • 1994
  • Casein or soybean protein was subjected to the reaction with caffeic acid-tyrosinase system at $30{\sim}35^{\circ}C$ pH 6.8 with aeration for 5 hr. The effects of the modified proteins on male Wistar rats were studied by pair-feeding of a cholesterol-free diet for 2 weeks. Significant decrease in protein digestibility for the rats fed with the modified proteins was observed. The feeding of modified protein from casein caused an enlargement of caecum. The concentration of serum cholesterol and triglyceride in the rats fed with modified proteins were mostly unchanged against the rats fed with untreated proteins. These results suggest that the decrease in protein digestibility induced by enzymic browning-reaction did not cause the decrease in concentration of serum cholesterol.

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