• Title/Summary/Keyword: inclusion bodies

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Comparison of Cytoplasmic Inclusions Induced by Maize Dwarf Mosaic Virus Strain A and B (Maize Dwarf Mosaic Virus strain A와 B에 의해 유도된 세포질 봉입체의 비교분석)

  • Choi, Chang-Won;Gardner, Wayne S.
    • Applied Microscopy
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    • v.24 no.2
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    • pp.105-114
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    • 1994
  • Comparative ultrastructural studies of sorghum (Sorghum bicolor L. Moench) cultivar (cv.) HOK and cv Pioneer 8680 leaf cells separately infected with maize dwarf mosaic virus (MDMV) strain A and B, respectively, revealed the formation of numerous cylindrical inclusions in both cross and longitudinal sections. The MDMV-A and -B were distinguished by the presence or absence of specific inclusion bodies in the cytoplasm. Electron microscope revealed the presence of pinwheels, bundles, scrolls, and laminated aggregates in Pioneer 8680 sorghum leaf cells infected with MDMV-B while no laminated aggregates were found in cells of HOK sorghum leaf cells infected with MDMV-A. Differences in the ultrastructure of cylindrical inclusions between two strains of MDMV, especially with respect to laminated aggregates, have been morphologically indexed to classify potyviruses into subdivision. The presence of laminated aggregates may be assigned to subdivision III while the absence of laminated aggregates is assigned to subdivision I. These variations of structures associated with cylindrical inclusions appeared virus-specific inductions and may be represented the morphogenesis of inclusion bodies following development of infections.

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In Vitro Formation of Protein Nanoparticle Using Recombinant Human Ferritin H and L Chains Produced from E. coli

  • RO HYEON SU;PARK HYUN KYU;KIM MIN GON;CHUNG BONG HYUN
    • Journal of Microbiology and Biotechnology
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    • v.15 no.2
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    • pp.254-258
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    • 2005
  • We have conducted in vitro reconstitution study of ferritin from its subunits FerH and FerL. For the reconstitution, FerH was produced from an expression vector construct in Escherichia coli and was purified from a heat treated cell extract by using one-step column chromatography. FerL was expressed as inclusion bodies. The denatured form of FerL was obtained by a simple washing step of the inclusion bodies with 3 M urea. The reconstitution experiment was conducted with various molar ratios of urea-denatured FerH and FerL to make the ferritin nanoparticle with a controlled composition of FerH and FerL. SDS-PAGE analysis of the reconstituted ferritins revealed that the reconstitution required the presence of more than 40 molar$\%$ of FerH in the reconstitution mixture. The assembly of the subunits into the ferritin nanoparticle was confmned by the presence of spherical particles with diameter of 10 nm by the atomic force microscopic image. Further analysis of the particles by using a transmission electron microscope revealed that the reconstituted particles exhibited different percentages of population with dense iron core. The reconstituted ferritin nanoparticles made with molar ratios of [FerH]/[FerL]=l00/0 and 60/40 showed that 80 to $90\%$ of the particles were apoferritin, devoid of iron core. On the contrary, all the particles formed with [FerH]/[FerL]=85/ 15 were found to contain the iron core. This suggests that although FerH can uptake iron, a minor portion of FerL, not exceeding $40\%$ at most, is required to deposit iron inside the particle.

Biological and Physicochemical Properties of Canine Parvovirus lated from the Dogs with Diarrhea (설사증 나환견으로 부터 분리한 Canine Parvovirus의 성상에 관한 연구)

  • 최해연;정운선;전무형;박성국;민원기
    • Korean Journal of Veterinary Service
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    • v.13 no.2
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    • pp.162-183
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    • 1990
  • From 1988 to 1989, 8 strains of canine parvovirus-2 (CPV-2) were isolated from the fecal specimens from the dogs that were clinically diagnosed as canine parvoviral enteritis in the veterinary hospitals located in the regions of Taejon and Chungbuk province. Studios on biological and physicochemical properties for the isolates were carried out. The results obtained by experiments are summarized as follows. 1. Among 62 fecal samples collected from the dogs with enteric diseases, 24 (38.7%) showed the haemagglutinating activity against porcine erythrocyte, ranging from 16 to 16, 384 of HA titers. 2. When 8 fecal specimens with high HA titer over 1, 000 were inocultated into CRFX cells, intranuclear inclusion bodies were obseverd in all of eight specimens, of w)lick three specimens showed cytoplasmic inclusions concurrently with the intranuclear inclusion bodies. 3. In study on species-specificity of haemagglutinating activity of the isolates, TJ-89-1 and TJ-89-2, it was found that the isolates revealed the highest haemagglutinating activity with porcine erythrocytes, showing the relatively lower haemagglutination titers with the erythrocytes from cat and rabbit. None of erythrocytes from other animals reacted with the isolates. 4. By the cross-haemagglutination inhibition test of the Isolates with reference viruses and sera, the Isolates were evidently identified as the strains of canine parvovirus-2. 5. In Physicochemical property test, it was evident that the isolates were stable in, lipid solvent, pH and heat treatment at $56^{\circ}C$ for 30 min. and contain DNA genome. 6. When seven puppies were inoculated intraorally with the isolate at HA titer of 8, 192, all of the puppies showed the symptoms of anorexia, vomiting, diarrhea and died at the 5th to 10th days post inoculation(pi). The fecal samples from all of the puppies revealed significantly high HA titers afterward the 5th days pi. Body temperature and the number of total leucocytes were slightly increased at the early stage of infection. but extremely decreased at the stage of collapse. HI titers of the sera started to increase at the 2nd to 3rd days pi reaching 512 to 1, 024 at the 4th to 5th day pi.

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Cloning, Expression, and Purification of a Lipase from Psychrotrophic Pseudomonas mandelii (Pseudomonas mandelii의 lipase 유전자 클로닝, 발현 및 정제)

  • Kim, Jun-Sung;Lee, Chang-Woo
    • Journal of Life Science
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    • v.22 no.3
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    • pp.306-311
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    • 2012
  • A gene encoding a lipase, lipT, was cloned from the psychrotrophic bacterium Pseudomonas mandelii and sequenced. An open reading frame of 1,686 bp was found that encodes a polypeptide consisting of 562 amino acid residues. Sequence analysis revealed a Gly-His-Ser-Leu-Gly sequence, which matches the consensus Gly-X-Ser-X-Gly motif conserved among lipolytic enzymes. The recombinant LipT protein was predominantly expressed as inclusion bodies in Escherichia coli and subsequently purified by nickel-chelate affinity chromatography. A small fraction of LipT was refolded, and the subsequent LipT exhibited substrate preferences for p-nitrophenyl butyrate (C4) and p-nitrophenyl octanoate (C8).

Pathological findings of the mixed infection with canine distemper virus and Streptococcus canis on farmed badger

  • Kim, Ji-hyeon;Lee, Kyunghyun;Jung, Ji-Youl;Choi, Eun-Jin;Kim, Ha-Young;So, ByungJae
    • Korean Journal of Veterinary Service
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    • v.41 no.1
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    • pp.51-55
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    • 2018
  • Herein, we report a case of badgers showing high morbidity and mortality rate due to the mixed infection of canine distemper virus (CDV) and Streptococcus canis (S. canis) in a farm where wild animal, badger, is being reared for herbal medicine. During the period of about one month, 120 out of 320 badgers showed severe respiratory symptoms and died, and 3 bodies were submitted to the Animal and Plant Quarantine Agency for disease diagnosis. The lung with the most severe necropsy findings failed to collapse and showed dark reddening and had yellowish nodules on the cut surface. The characteristic and common histopathologic findings include multifocal necrosis with hemorrhage of the lung, severe lymphoid depletion of the spleen and intracytoplasmic or intranuclear inclusion bodies in almost all organs. Finally, CDV and S. canis were identified by immunohistochemistry and bacterial isolation, respectively. This is the first mixed infection case of CDV and S. canis in badgers being raised on the farm.

A Study on the Synthesis of Anorthite and its Characteristics. (Anorthite의 합성 및 그 특성에 관한 연구)

  • 백용혁;이종권
    • Journal of the Korean Ceramic Society
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    • v.20 no.2
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    • pp.153-160
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    • 1983
  • This study was carried out to research the change of mineral phases and the characteristics(apparent specific gra-vity water absorption firing shrinkage modulus of rupture thermal expansion and specific dielectric constant) of the sintered bodies manufactured by kaolin and limestone. Samples were composed of the same theoretical composition as it of anorthite and fired up to 145$0^{\circ}C$ Investigated the change and micro-structure of the mineral phases by XRD and SEM the characterisdtics of the sintered bodies by TMA Automatic Capacitance Bridge and etc. The results were as follow. 1. Reactions of sintering are occurred between 860-95$0^{\circ}C$ and 1200-138$0^{\circ}C$ and state of bloating is occurred at 1410-145$0^{\circ}C$ 2. For the inclusion of feldspar and its fine particles of materials the temperature of producing and collapsing is decreased. 3. Pseudo-wollastonite and gehlenite are formed about 95$0^{\circ}C$ 4. At 114$0^{\circ}C$ anorthite are begin to forming and increase continuously to 138$0^{\circ}C$. Above 141$0^{\circ}C$ content of anorthite are decreased. 5. The variations of bending strength with sintering temperature reflect similar trend of sintered contraction and in-crease continuously from 120$0^{\circ}C$. At 145$0^{\circ}C$ reached about 680kg/cm2. 6. Specific dielectric constant$($\varepsilon$_s)$ of specimen sintered at 141$0^{\circ}C$ is 7.12 and that value is most favorable.

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Identification of Bean Common Mosaic Virus Obtained from Seeds of Phaseolus vulgaris (강낭콩에서 종자전염된 Bean Common Mosaic Virus의 분류동정에 관한 연구)

  • Choi Y.M.;Lee S.H.;Park J.S.;Kim J.S.
    • Korean journal of applied entomology
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    • v.23 no.1 s.58
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    • pp.15-21
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    • 1984
  • The virus infecting French bean (Phaseolus vulgaris L.) was identified as Bean Common Mosaic Virus(BCMV) based on the host range, symptomatology, serology, morphology of virus particles and inclusion bodies. Isolates of BCMV were obtained from seeds of P. vulgaris collected at Suweon, Jangsu and Jinju in Korea. French bean produced vein clearing, mosaic, stunting and leaf curling. Symptom of Chenopodium quinoa was local lesions on the inoculated leaves, not on the upper leaves. The electron micrograph of the virus from French bean was flexuous approximately 750nm in length. Cylindrical and pinwheel cytoplasmic inclusion bodies were observed in French bean leaf infected by BCMV. BCMV from the French bean was transmitted through seed and green peach aphid, Myzus persicae. The thermal inactivation point was $55\~60^{\circ}C$, dilution end point was $10^{-3}\~10^{-5}$ and longevity in vitro was $2\~3$ days for BCMV from French bean. The isolates of BCMV reacted positively against BCMV antiserum. The extract of BCMV infected bean leaves, Azukibean mosaic virus (AZMV) and Cowpea aphid borne mosaic virus(CaMV) also reacted with BCMV antiserum, however, BCMV and CaMV showed the spur in agar gel diffusion test.

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Addition of an N-Terminal Poly-Glutamate Fusion Tag Improves Solubility and Production of Recombinant TAT-Cre Recombinase in Escherichia coli

  • Kim, A-Hyeon;Lee, Soohyun;Jeon, Suwon;Kim, Goon-Tae;Lee, Eun Jig;Kim, Daham;Kim, Younggyu;Park, Tae-Sik
    • Journal of Microbiology and Biotechnology
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    • v.30 no.1
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    • pp.109-117
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    • 2020
  • Cre recombinase is widely used to manipulate DNA sequences for both in vitro and in vivo research. Attachment of a trans-activator of transcription (TAT) sequence to Cre allows TAT-Cre to penetrate the cell membrane, and the addition of a nuclear localization signal (NLS) helps the enzyme to translocate into the nucleus. Since the yield of recombinant TAT-Cre is limited by formation of inclusion bodies, we hypothesized that the positively charged arginine-rich TAT sequence causes the inclusion body formation, whereas its neutralization by the addition of a negatively charged sequence improves solubility of the protein. To prove this, we neutralized the positively charged TAT sequence by proximally attaching a negatively charged poly-glutamate (E12) sequence. We found that the E12 tag improved the solubility and yield of E12-TAT-NLS-Cre (E12-TAT-Cre) compared with those of TAT-NLS-Cre (TAT-Cre) when expressed in E. coli. Furthermore, the growth of cells expressing E12-TAT-Cre was increased compared with that of the cells expressing TAT-Cre. Efficacy of the purified TAT-Cre was confirmed by a recombination test on a floxed plasmid in a cell-free system and 293 FT cells. Taken together, our results suggest that attachment of the E12 sequence to TAT-Cre improves its solubility during expression in E. coli (possibly by neutralizing the ionic-charge effects of the TAT sequence) and consequently increases the yield. This method can be applied to the production of transducible proteins for research and therapeutic purposes.

Body discourse on DE&I in the fashion industry analyzed through The New York Times (뉴욕타임즈를 통해 분석한 패션산업 내 DE&I에 관한 신체담론)

  • Myeongseon Yi;Eunhyuk Yim
    • The Research Journal of the Costume Culture
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    • v.32 no.2
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    • pp.164-180
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    • 2024
  • In the context of a globalized society where diversity, equity, and inclusion (DE&I) have emerged as pivotal values, the fashion industry is undergoing scrutiny for its practices related to body DE&I. This study examines the nature of the discourse surrounding body DE&I within the fashion industry, focusing on how such discussions are shaped, disseminated, and manifested in both the industry and broader society. Critical discourse analysis is applied by utilizing, content from the New York Times and leveraging Fairclough's analytical framework encompassing textual, discursive, and social practices. The findings indicate that the New York Times emphasizes diversity, with a significant focus on the shapes and sizes of women's bodies, developing a narrative centered around women's bodies through visible and representative domains. The analysis suggests conflicted discourse, with prevailing critiques against the fashion industry's standardization of beauty and superficial inclusivity efforts. Moreover, the industry's adaptation to social demands for body DE&I is observed as sporadic, often leveraging non-normative bodies as a marketing strategy rather than genuinely embracing diversity. This study highlights the importance of continuous, in-depth discourse and social practices regarding DE&I within the fashion industry, as well as the need for systemic changes and policies that genuinely reflect societal demands for inclusivity. The findings provide a foundation for future investigations into the multifaceted relationship between fashion discourse, DE&I, and social practices, advocating for a more inclusive and critically aware fashion industry.

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
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    • v.6 no.3
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    • pp.162-166
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    • 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.

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