• 제목/요약/키워드: Protein bodies

검색결과 326건 처리시간 0.032초

화경버섯의 항세균성 렉틴 (An Antibacterial Lectin from Lampteromyces japonicus)

  • 윤주억;민태진;윤희식
    • 한국균학회지
    • /
    • 제23권1호통권72호
    • /
    • pp.46-52
    • /
    • 1995
  • 화경버섯의 자실체로부터 PAGE로 한 렉틴을 정제하고, LJAP(Lampteromyces japonicus antibacterial protein)라 이름 붙였다. LJAP는 17-kDa의 서브유닛트가 모여서 된 백 kDa이 넘는 회합 단백질이였다. 이 단백질은 아미노산 분석결과 세린, 글리신, 산성 아미노산의 함량이 높았다. LJAP는 대장균중 특히 JM 109, K 12, HB 101, JW 380에 대한 항세균작용이 뛰어났다. 한편, 항세균활성은 당단백질인 asialofetuin에 의하여 크게 억제되었다. 본 당단백질은 화경버섯에서 처음으로 얻어진 항세균활성과 적혈구 응집활성을 가진 렉틴이다.

  • PDF

Development of a Recombinant Protein Vaccine Based on Cell-Free Protein Synthesis for Sevenband Grouper Epinephelus septemfasciatus Against Viral Nervous Necrosis

  • Kim, Jong-Oh;Kim, Jae-Ok;Kim, Wi-Sik;Oh, Myung-Joo
    • Journal of Microbiology and Biotechnology
    • /
    • 제25권10호
    • /
    • pp.1761-1767
    • /
    • 2015
  • Sevenband grouper, Epinephelus septemfasciatus, is becoming an important aquaculture species in Korea. However, viral nervous necrosis disease is a large problem causing mass mortality in sevenband grouper aquaculture. Recombinant protein vaccines are one of the best methods to reduce these economic losses. However, the cell-based expression method mainly produces inclusion bodies and requires additional procedures. In this study, we expressed a recombinant viral coat protein of sevenband grouper nervous necrosis virus (NNV) using a cell-free protein synthesis system. The purified recombinant NNV coat protein (rNNV-CP) was injected into sevenband grouper at different doses followed by a NNV challenge. Nonimmunized fish in the first trial (20 μg/fish) began to die 5 days post-challenge and reached 70% cumulative mortality. In contrast, immunized fish also starting dying 5 days postchallenge but lower cumulative mortality (10%) was observed. Cumulative morality in the second trial with different doses (20, 4, and 0.8 μg/fish) was 10%, 40%, and 50%, respectively. These results suggest that rNNV-CP can effectively immunize sevenband grouper depending on the dose administered. This study provides a new approach to develop a recombinant vaccine against NNV infection for sevenband grouper.

High-level Expression and Purification of Recombinant 4-Aminobutyrate Aminotransferases in Escherichia coli

  • Lee, Sung Gu;Tae Jin Choi;Young Tae Kim
    • Journal of Microbiology and Biotechnology
    • /
    • 제6권3호
    • /
    • pp.162-166
    • /
    • 1996
  • The protein coding sequence of the 4-aminobutyrate aminotransferase was amplified by polymerase chain reaction (PCR) from a previously cloned cDNA of pig brain using a pair of primers based on the published sequence. The amplified DNA was introduced into a T7 expression vector. Recombinant 4-aminobutyrate aminotransferases were overexpressed in Escherichia coli. The inclusion bodies were formed when enzyme was overexpressed. The unfolded, overproduced proteins were purified by chromatography with hydroxyapatite and refolded by a sequential dialysis method. The renatured 4-aminobutyrate aminotransferase regained the catalytic activity. However, the purified mutant protein did not show the catalytic function of 4-aminobutyrate aminotransferase.

  • PDF

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

  • SeungYup Lee;Jeo
    • Biotechnology and Bioprocess Engineering:BBE
    • /
    • 제1권1호
    • /
    • pp.9-12
    • /
    • 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.

  • PDF

A Unique Strategy for Recovering Recombinant Proteins from Molecular Farming: Affinity Couture on Engineered Oilbodies

  • Seon, Jeong-Hoon;J.Steven Szarka;Maurice M. Moloney
    • Journal of Plant Biotechnology
    • /
    • 제4권3호
    • /
    • pp.95-101
    • /
    • 2002
  • Molecular faming has the potential to provide large amounts of recombinant protein for use in diagnostics and as therapeutics. Various strategies have been developed to enhance the expression level, stability, and native folding of recombinant proteins produced in plants. Few investigations into the subcellular distribution of recombinant proteins within plant cells have been published despite the potential to increase the expression level and impact the purification process. This review article discusses the current strategies used for targeting recombinant proteins to various subcellular locations and the advantages of targeting to seed oil bodies for molecular farming applications. Specifically, the affinity capture of antibodies using recombinant oilbodies is discussed.

Microscopy of Microbial Gas Vesicles

  • Park, Junhyung;Kim, Ki Woo
    • Applied Microscopy
    • /
    • 제47권3호
    • /
    • pp.165-170
    • /
    • 2017
  • Gas vesicles are intracellular gas-filled protein-shelled nanocompartments. The structures are spindle or cylinder-shaped, and typically $0.1{\sim}2{\mu}m$ in length and 45~250 nm in width. A variety of prokaryotes including photosynthetic bacteria and halophilic archaea form gas vesicles in their cytoplasm. Gas vesicles provide cell buoyancy as flotation devices in aqueous habitats. They are used as nanoscale molecular reporters for ultrasound imaging for biomedical purposes. The structures in halophilic archaea are poorly resolved due to the low signal-to-noise ratio from the high salt concentration in the medium. Such a limitation can be overcome using focused ion beam-thinning or inelastically scattered electrons. As the concentric bodies (~200 nm in diameter) in fungi possess gas-filled cores, it is possible that the concept of gas vesicles could be applied to eukaryotic microbes beyond prokaryotes.

Genome wide identification of Staufen2-bound mRNAs in embryonic rat brains

  • Maher-Laporte, Marjolaine;DesGroseillers, Luc
    • BMB Reports
    • /
    • 제43권5호
    • /
    • pp.344-348
    • /
    • 2010
  • Messenger ribonucleoprotein particles (mRNPs) are used to transport mRNAs along neuronal dendrites to their site of translation. Staufen2 is an mRNA-binding protein expressed in the cell bodies and cellular processes of different brain cells. It is notably involved in the transport of dendritic mRNAs along microtubules. Its knockdown expression was shown to change spine morphology and impair synaptic functions. However, the identity of Staufen2-bound mRNAs in brain cells is still completely unknown. As a mean to identify these mRNAs, we immunoprecipitated Staufen2-containing mRNPs from embryonic rat brains and used a genome wide approach to identify Staufen2-associated mRNAs. The genome wide approach identified 1780 mRNAs in Staufen2-containing mRNPs that code for proteins involved in cellular processes such as post-translational protein modifications, RNA metabolism, intracellular transport and translation. These results represent an additional and important step in the characterization of Staufen2- mediated neuronal functions in rat brains.

인삼(Panax ginseng C.A. Meyer)의 종자형성에 있어서 배유세포의 미세구조의 변화 및 저장물질의 형성 (Ultrastructural Changes and Formation of Storage Materials in Endosperm Cells during the Seed Formation of Panax ginseng C.A. Meyer)

  • 유성철
    • Journal of Plant Biology
    • /
    • 제34권3호
    • /
    • pp.201-213
    • /
    • 1991
  • This study has been carried out to investigate the ultrastructural changes, formation of storage materials in endosperm cells with electron microscope during the seed formation of Panax ginseng C.A. Meyer. In the early stage of seed formation with green seed coat, the endosperm was cellular type. Cell plate was largely composed of dictyosome vesicles in early stage of wall formation after mitosis. Central vacuole was gradually subdivided into several small-sized vacuoles. During the differentiation of plastids, some proplastid was replaced by amyloplast with starch grains and lamellar structure. A number of mitochondria with well developed cristae were distributed in cytoplasm. Rough endoplasmc reticulum, dictyosome, microbody, free ribosomes and polysomes were evenly distributed in cytoplasm. Spherical spherosomes were formed from dictyosome containing the lipid materials of even electron density. Protein bodies were formed by interfusing between vacuoles and vesicles derived from rough endoplasmic reticulum which contained the amorphous protein of high electron density.

  • PDF