• Title/Summary/Keyword: Protein bodies

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The First Report of a Megalocytivirus Infection in Farmed Starry Flounder, Platichthys stellatus, in Korea

  • Won, Kyoung-Mi;Cho, Mi Young;Park, Myoung Ae;Jee, Bo Young;Myeong, Jeong-In;Kim, Jin Woo
    • Fisheries and Aquatic Sciences
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    • v.16 no.2
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    • pp.93-99
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    • 2013
  • In 2009, a systemic megalocytivirus infection associated with high mortality was detected for the first time in cultured starry flounder Platichthys stellatus in Korea. Diseased starry flounder had pale bodies and gill coloring and enlarged spleens. Histopathological examinations revealed basophilic enlarged cells in various organs of diseased starry flounder. Polymerase chain reaction (PCR) was performed on tissue samples using three published primer sets developed for the red sea bream iridovirus. PCR products were detected for all primer sets, except 1-F/1-R, which are registered by the World Organization for Animal Health (OIE). The part of the gene corresponding to the full open reading frame encoding the viral major capsid protein (MCP) was amplified by PCR. PCR products of approximately 1,581 bp were cloned, and the nucleotide sequences were analyzed phylogenetically. The MCP gene of the starry flounder iridovirus, designated SFIV0909, was identical to that of the turbot reddish body iridovirus (AB166788).

Inhibition of Cell Proliferation and Induction of Apoptosis by Methanolic Extract of Aconiti Tuber in Human Renal Cell Carcinoma Cells (부자의 메탄올 추출물에 의한 신세포암의 증식억제 및 세포사멸 유도)

  • Kim, Jong-Hwan;Choi, Yung-Hyun;Hwang, Won-Deuk
    • The Journal of Internal Korean Medicine
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    • v.29 no.1
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    • pp.42-66
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    • 2008
  • Aconiti Tuber is a traditional medicinal plant generally used in Oriental medicine therapy. In this study, we investigated the biochemical mechanisms of anti-proliferative effects by the methanol extract of Aconiti tuber (MEBJ) in Caki-1 human renal cell carcinoma cells. It was found that MEBJ could inhibit, in a dose-dependent manner, cell growth which was associated with apoptotic cell death such as formation of apoptotic bodies, DNA fragmentation and increased populations of apoptotic-sub G1 phase. Apoptosis of Caki-1 cells by MEBJ was associated with an up-regulation of pro-apoptotic Bax expression, and a down-regulation of anti-apoptotic Bcl-2 in a dose-dependent manner; however, the levels of IAP family were not affected. MEBJ treatment also induced the proteolytic activation of caspase-3 and -8, and a inhibition of poly(ADP-ribose) polymerase (PARP) and $PLC{\gamma}1$ protein. Furthermore, MEBJ treatment caused a dose-dependent inhibition of iNOS and cyclooxygenase-2 (Cox-2). Though further studies will be needed to identify the active compounds that confer the anti-cancer activity of MEBJ, the present findings provide important new insights into the possible molecular mechanisms of the apoptotic activity of MEBJ in cancer cells.

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Microstructure of Lupin Seed;a Comparative Study With Soybean (루핀콩과 대두의 미세구조에 관한 비교 연구)

  • Lee, Cherl-Ho;Kim, Jeong-Kyo
    • Korean Journal of Food Science and Technology
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    • v.17 no.6
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    • pp.454-459
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    • 1985
  • The structure of the seed of Lupinue angustifolius was studied in order to investigate the Food quality of lupin seed in comparision with soybean. The cotyledonary cells of lupinseed was in egg-like shape and much (more than 4 times) larger than those of soybean. The microstructure of cotyledonary cells of lupinseed was characterized with thick cell wall having distinct pit-pairs. The protein bodies in lupinseed cotyledon cell contained numerous crystaloids, which was absent in soybean. The middle lamella of soybean cell was partially disintegrated by excessive heat treatment ($120^{\circ}C$, 20 min), whereas those of lupinseed did not change much by heting at $120^{\circ}C$ for 130 min.

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Expression and Purification of Recombinant Active Prostate-Specific Antigen from Escherichia coli

  • Jeong, Su-Jin;Lee, Seong-Wook
    • Journal of Microbiology and Biotechnology
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    • v.17 no.5
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    • pp.840-846
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    • 2007
  • Human prostate-specific antigen(PSA), a 33 kDa serine protease with comprehensive homology to glandular kallikrein, is secreted from prostatic tissue into the seminal fluid and enters into the circulation. The level of PSA increases in the serum of patients with prostatic cancer and hence is widely employed as a marker of the disease status. In particular, an enzymatically active PSA that is a form cleaved at the N-terminal seven-amino-acids prosequence, APLILSR, of proPSA may play an important roll in the progression of prostate cancer. Thus, the presence of the active form would selectively discriminate the cancer from benign prostatic hyperplasia. In this study, we developed a convenient purification method for the acquisition of active PSA and proPSA. Recombinant proPSA and active PSA were expressed directly in Escherichia coli, easily and efficiently isolated from inclusion bodies, refolded, and purified. Moreover, the enzymatic activity of the recombinant active PSA was confirmed as serine protease using chromogenic chymotrypsin substrate. This purified active PSA could be further applied to scrutinize the biological or conformational characteristics of the protein and to develop specific diagnostic and/or therapeutic agents against prostate cancer.

Escherichia coli Cytoplasmic Expression of Disulfide-Bonded Proteins: Side-by-Side Comparison between Two Competing Strategies

  • Angel Castillo-Corujo;Yuko Uchida;Mirva J. Saaranen;Lloyd W. Ruddock
    • Journal of Microbiology and Biotechnology
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    • v.34 no.5
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    • pp.1126-1134
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    • 2024
  • The production of disulfide bond-containing recombinant proteins in Escherichia coli has traditionally been done by either refolding from inclusion bodies or by targeting the protein to the periplasm. However, both approaches have limitations. Two broad strategies were developed to allow the production of proteins with disulfide bonds in the cytoplasm of E. coli: i) engineered strains with deletions in the disulfide reduction pathways, e.g. SHuffle, and ii) the co-expression of oxidative folding catalysts, e.g. CyDisCo. However, to our knowledge, the relative effectiveness of these strategies has not been properly evaluated. Here, we systematically compare the purified yields of 14 different proteins of interest (POI) that contain disulfide bonds in their native state when expressed in both systems. We also compared the effects of different background strains, commonly used promoters, and two media types: defined and rich autoinduction. In rich autoinduction media, POI which can be produced in a soluble (non-native) state without a system for disulfide bond formation were produced in higher purified yields from SHuffle, whereas all other proteins were produced in higher purified yields using CyDisCo. In chemically defined media, purified yields were at least 10x higher in all cases using CyDisCo. In addition, the quality of the three POI tested was superior when produced using CyDisCo.

Ataxin-2 Dysregulation Triggers a Compensatory Fragile X Mental Retardation Protein Decrease in Drosophila C4da Neurons

  • Cha, In Jun;Lee, Davin;Park, Sung Soon;Chung, Chang Geon;Kim, Seung Yeon;Jo, Min Gu;Kim, Seung Yeol;Lee, Byung-Hoon;Lee, Young-Sam;Lee, Sung Bae
    • Molecules and Cells
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    • v.43 no.10
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    • pp.870-879
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    • 2020
  • Dendrites require precise and timely delivery of protein substrates to distal areas to ensure the correct morphology and function of neurons. Many of these protein substrates are supplied in the form of ribonucleoprotein (RNP) complex consisting of RNA-binding proteins (RBPs) and mRNAs, which are subsequently translated in distal dendritic areas. It remains elusive, however, whether key RBPs supply mRNA according to local demands individually or in a coordinated manner. In this study, we investigated how Drosophila sensory neurons respond to the dysregulation of a disease-associated RBP, Ataxin-2 (ATX2), which leads to dendritic defects. We found that ATX2 plays a crucial role in spacing dendritic branches for the optimal dendritic receptive fields in Drosophila class IV dendritic arborization (C4da) neurons, where both expression level and subcellular location of ATX2 contribute significantly to this effect. We showed that translational upregulation through the expression of eukaryotic translation initiation factor 4E (eIF4E) further enhanced the ATX2-induced dendritic phenotypes. Additionally, we found that the expression level of another disease-associated RBP, fragile X mental retardation protein (FMRP), decreased in both cell bodies and dendrites when neurons were faced with aberrant upregulation of ATX2. Finally, we revealed that the PAM2 motif of ATX2, which mediates its interaction with poly(A)-binding protein (PABP), is potentially necessary for the decrease of FMRP in certain neuronal stress conditions. Collectively, our data suggest that dysregulation of RBPs triggers a compensatory regulation of other functionally-overlapping RBPs to minimize RBP dysregulation-associated aberrations that hinder neuronal homeostasis in dendrites.

Biosynthesis of Organic Germanium Using Cordyceps militaris (동충하초를 이용한 유기게르마늄의 생산)

  • Kim, Seung;Kim, Jae-Sung;Sapkota, Kumar;Choi, Bong-Suk;Park, Se-Eun;Park, Yeal;Chun, Hong-Sung;Yoo, Jin-Cheol;Choi, Han-Seok;Kim, Myung-Kon;Kim, Sung-Jun
    • The Korean Journal of Mycology
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    • v.34 no.2
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    • pp.84-87
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    • 2006
  • In the present study, an attempt has been made to produce a high quality medicinal mushroom Cordycops militaris supplemented with organic Ge. Cordycops militaris was cultivated in SDAY liquid medium containing yeast extract 10 g, peptone 10 g, glucose 40 g per liter and chrysalis media supplemented with inorganic Ge at 100, 500, 1000, and 5000 ppm concentrations. The greatest organic Ge production of 442.4 ppm/g and 284 ppm/g were observed in SDAY liquid and Chrysalis media cultures supplemented with 100 ppm inorganic Ge respectively. Similarly, 4,509.7 ppm/g and 1,058 ppm/g of organic Ge were obtained from liquid and chrysalis media cultures supplemented with 5,000 ppm and 1,000 ppm inorganic Ge, respectively. In addition, higher concentration of organic Ge was obtained in mycelia than fruiting bodies. These results indicate that the concentration of organic Ge increase with decreasing inorganic Ge concentration in the medium. This is the first report on production of high valuable Cordyceps militaris contained with organic Ge.

Parasporin-4, A Novel Cancer Cell-killing Protein Produced by Bacillus thuringiensis

  • Inouye, Kuniyo;Okumura, Shiro;Mizuki, Eiichi
    • Food Science and Biotechnology
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    • v.17 no.2
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    • pp.219-227
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    • 2008
  • Bacillus thuringiensis was isolated as a pathogen of the sotto disease of silkmoth larvae about a hundred years ago. Since then, this bacterium has attracted attentions of not only insect pathologists but also many other scientists who are interested in its strong and specific insecticidal activity. This has led to the recent worldwide development of B. thuringiensis-based microbial insecticides and insect-resistant transgenic plants, as well as a landmark discovery of par asp orin, a cancer cell-specific cytotoxin produced by B. thuringiensis. In this review, we describe examination of interaction between inclusion proteins of B. thuringiensis and brush border membrane of insects using a surface plasmon resonance-based biosensor, identification and characterization of parasporin-4, the latest parasporin produced by the B. thuringiensis A1470 strain, and an effective method for preparing the parasporin-4 from inclusion bodies expressed in the recombinant Escherichia coli cells.

Localization and Function of Cellulase in Endosperm Cells of Panax ginseng Seeds during Maturation and After-ripening (인삼 종자의 성숙과 후숙 과정에서 배유세포내 섬유소 가수분해효소의 분포 및 기능)

  • 유성철
    • Journal of Plant Biology
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    • v.36 no.4
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    • pp.327-335
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    • 1993
  • The active sites, intracellular transport, function of cellulase in association with the disintegration of the storage materials of the endosperm cells during seed maturation and after-ripening of Panax ginseng C.A. Meyer seeds were studied by electron microscopy. Cytochemical activities of the cellulase occurred in protein bodies and vesicles of endosperm cells in seed with red seed coat. In after-ripening seed, the activities were strongly found in the cell wall of endosperm near the umbiliform layer and on neighbouring vesicles, so it is assumed that these cells begin to be decomposed. Cellulase activities were initiated before the decomposition of storage materials. But, no activity was observed in the umbiliform layer, so it is suggested that cellulase lose its activity after the completion of lysis process.

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Effects of Aloe vera Treatment on Blood Glucose Level and Clinical Chemistry in Diabetic Patients (당뇨병 환자의 혈당 및 임상에 미치는 알로에의 효과)

  • 박웅양;윤여표
    • Journal of Food Hygiene and Safety
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    • v.10 no.1
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    • pp.13-17
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    • 1995
  • In order to study the effects of Aloe vera treatment on blood glucose level and clinical chemistry in diabetic patients, eight diabetic patients were administered orally with 800 mg of Aloe vera three time a day for three months. The high levels of blood and urine glucose in diabetic patients were significantly reduced by administration of Aloe vera. The increased plasma triglyceride concentration was also significantly reduced by Aloe vera treatment. A little amount of urine bilirubin, hematuria, nitrite, urobilinogen, protein and ketone bodies were detected before treatment, but not detected after Aloe vera treatment. But other blood parameters of clinical chemistry values were not affected by Aloe vera treatment. These data suggest that Aloe vera can be effective in the treatment of the diabetic patients.

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