• Title/Summary/Keyword: gels

Search Result 730, Processing Time 0.029 seconds

Comparative proteome analysis of diploid and tetraploid root in Platycodon grandiflorum

  • Kwon, Soo Jeong;Roy, Swapan Kumar;Yoo, Jang-Hawan;Cho, Seong-Woo;Kim, Hag Hyun;Boo, Hee Ock;Woo, Sun-Hee
    • Proceedings of the Korean Society of Crop Science Conference
    • /
    • 2017.06a
    • /
    • pp.123-123
    • /
    • 2017
  • In spite of the potential medicinal significance and a wide range of pharmacologic properties of Platycodon grandiflorum, the molecular mechanism of its roots is still unknown. The present study was conducted to profile proteins from 3, 4 and 5 months aged diploid and tetraploid roots of Platycodon grandiflorum using high throughput proteome approach. Two-dimensional gels stained with CBB, a total of 68 differential expressed proteins were identified from the diploid root out of 767 protein spots using image analysis by Progenesis SameSpot software. Out of total differential expressed spots, 29 differential expressed protein spots (${\geq}2-fold$) were analyzed using LTQ-FTICR MS whereas a total of 24 protein spots were up-regulated and 5 protein spots were down-regulated. On the contrary, in the case of tetraploid root, a total of 86 differential expressed proteins were identified from tetraploid root out of 1033 protein spots of which a total of 39 differential expressed protein spots (${\geq}2-fold$) were analyzed using LTQ-FTICR MS whereas a total of 21 protein spots were up-regulated and a total of 18 protein spots were down-regulated. It was revealed that the identified proteins from the explants were mainly associated with the nucleotide binding, oxidoreductase activity, transferase activity. Taken together, the identified proteins may be helpful to identify key candidate proteins for genetic improvement of plants.

  • PDF

Proteome characterization of the liquid cultured tetraploid roots in Platycodon grandiflorum

  • Ko, Jung-Hee;Kwon, Soo Jeong;Roy, Swapan Kumar;Cho, Seong-Woo;Kim, Hag Hyun;Boo, Hee Ock;Woo, Sun-Hee
    • Proceedings of the Korean Society of Crop Science Conference
    • /
    • 2017.06a
    • /
    • pp.125-125
    • /
    • 2017
  • The roots of Platycodon grandiflorum are commonly used for treating bronchitis, asthma, tuberculosis, diabetes, and other inflammatory diseases. Since the molecular mechanism underlying the roots of the plant is unclear. Therefore, the present study was conducted to profile proteins from liquid cultured tetraploid roots of Platycodon grandi orum fl using high throughput proteome approach. Two-dimensional gels stained with CBB, a total of 659 differentially expressed proteins were identified from the liquid medium cultured tetraploid roots of which 32 proteins spots (${\geq}1.5-fold$) were sorted for mass spectrometry analysis. Out of these 32 proteins, a total of 15 proteins were up-regulated such as Serine carboxypeptidase-like 27, Transcription factor bHLH150, 60 kDa jasmonate-induced protein, Cytosolic Fe-S cluster assembly factor NBP35, Regulatory associated protein of TOR 2 and a total of 17 proteins were down-regulated such as Protein G1-like2, Phenylalanine ammonia-lyase, Fructokinase-2, Trihelix transcription factor GT-3a, Guanine nucleotide-binding protein alpha-1 subunit. However, the frequency distribution of identified proteins was carried out within functional categories based on molecular functions, cellular components, and biological processes. Functional categorization revealed that the most of the identified proteins from the explants were mainly associated with the nucleic acid binding, oxidoreductase, transferase activity, protein binding and hydrolase activity. In addition, the proteomic feedback of tetraploid roots of P. grandiflorum may potentially be used to understand the characteristics of proteins and their functions.

  • PDF

Expression and Antigenicity of Replicase Protein from Snow Mountain-Like Caliciviruses, Korean Isolates (한국형 사람 Calicivirus Replicase 단백의 발현 및 항원성 평가)

  • Chang, Mi-Yoon;Yang, Jai-Myung;Kim, Kyung-Hee
    • The Journal of Korean Society of Virology
    • /
    • v.27 no.2
    • /
    • pp.151-160
    • /
    • 1997
  • In view of the potential of replicase protein as a diagnostic reagent for human caliciviruses (HuCVs), we have cloned and over-expressed this gene from the Snow Mountain-like Korean strains in Escherichia coli as a fusion protein with glutathione S-transferase (GST), and described the preliminary antigenic characterization of the recombinant products. Each 470bp fragment corresponding to highly conserved region of RNA-dependent RNA polymerase was generated by RT-PCR from stools of two diarrheal children, cloned in pMOSBlue T-vector, and subcloned between the EcoRI and SalI restriction sites of pGEX-4T-3, a GST gene fusion vector, yielding $pGCV_{pol}$. This construct expressed a Snow Mountain-like HuCV replicase under the control of the IPTG-inducible tac promoter. An extract prepared by sonication of the E. coli cell inclusion bodies bearing $pGCV_{pol}$ products was purified and analyzed by SDS-PAGE. After Coomassie blue staining, it was shown that the recombinant replicase migrated on the gels with an approximate molecular mass of 46.5 kDa, that was subsequently cleaved into a 26 kDa GST fragment and a 20.5 kDa replicase protein upon digestion with thrombin protease. The replicase was recognized on immunoblotting with the sera from symptomatic children with the HuCV-associated diarrhea but not by asymptomatic sera from adults. The results presented the first biological activity of individually expressed HuCV replicase subunit and provided important reagents for diagnosis of HuCV infection.

  • PDF

Effect of Glycolysis Rate in Porcine Muscle Postmortem on Gel Property of Pork Surimi (돼지 근육의 사후 해당속도가 돈육 수리미의 젤 특성에 미치는 영향)

  • Kang Guen-Ho;Yang Han-Sul;Jeong Jin-Yeon;Joo Seon-Tea;Park Gu-Boo
    • Food Science of Animal Resources
    • /
    • v.25 no.4
    • /
    • pp.423-429
    • /
    • 2005
  • Properties of pant surimi derived from porcine longissimus muscle were investigated Rapid glycolysis of muscle reduced yield $\%$ of water-washed pork and moisture $\%$ of pent surimi because of ie lower ultimate pH. Gel Hardness was significantly (p<0.05) higher in pork surimi from rapid glycolysis muscle, but springiness was higher (p<0.05) in pork surimi from normal glycolysis muscle. SDS-PAGE pattern showed denaturation of sarcoplasmic proteins onto myofibrillar proteins in rapid glycolysis muscle, resulted in dark color and hard texture of pork surimi. Color and texture of gels were related with water-holding capacity of muscle proteins and moisture $\%$ in gel matrix. Results imply that glycolysis rate of porcine muscle at postmortem could affect gel properties of pork surimi, and muscle with rapid glycolysis muscle could produce a hard texture of pork surimi and dark color.

Evaluation of Acid-treated Fish Sarcoplasmic Proteins on Physicochemical and Rheological Characteristics of Pork Myofibrillar Protein Gel Mediated by Microbial Transglutaminase

  • Hemung, Bung-Orn;Chin, Koo Bok
    • Food Science of Animal Resources
    • /
    • v.35 no.1
    • /
    • pp.50-57
    • /
    • 2015
  • Fish sarcoplasmic protein (SP) is currently dumped as waste from surimi industry and its recovery by practical method for being the non-meat ingredient in meat industry would be a strategy to utilize effectively the fish resource. This study was aimed to apply pH treatment for fish SP recovery and evaluated its effect on pork myofibrillar protein (MP) gel. The pH values of fish SP were changed to 3 and 12, and neutralized to pH 7 before lyophilizing the precipitated protein after centrifugation. Acid-treated fish SP (AFSP) showed about 4-fold higher recovery yield than that of alkaline-treated SP and water absorption capacity was also about 1.2-fold greater. Because of the high recovery yield and water absorption capacity, AFSP was selected to incorporate into MP with/without microbial transglutaminase (MTG). The effects of AFSP and MTG on the physicochemical and rheological characteristics of MP and MP gel were evaluated. MTG induced an increase shear stress of the MP mixture and increase the breaking force of MP gels. MP gel lightness was decreased by adding AFSP. MP gel with MTG showed higher cooking loss than that without MTG. A reduction of cooking loss was observed when the AFSP was added along with MTG, where the insoluble particles were found. Therefore, AFSP could be contributed as a water holding agent in meat protein gel.

Effects of subsequent curing on chloride resistance and microstructure of steam-cured mortar

  • Hu, Yuquan;Hu, Shaowei;Yang, Bokai;Wang, Siyao
    • Advances in concrete construction
    • /
    • v.9 no.5
    • /
    • pp.449-457
    • /
    • 2020
  • The influence of subsequent curing on the performance of fly ash contained mortar under steam curing was studied. Mortar samples incorporated with different content (0%, 20%, 50% and 70%) of Class F fly ash under five typical subsequent curing conditions, including standard curing (ZS), water curing(ZW) under 25℃, oven-dry curing (ZD) under 60℃, frozen curing (ZF) under -10℃, and nature curing (ZN) exposed to outdoor environment were implemented. The unsteady chloride diffusion coefficient was measured by rapid chloride migration test (RCM) to analyze the influence of subsequent curing condition on the resistance to chloride penetration of fly ash contained mortar under steam curing. The compressive strength was measured to analyze the mechanical properties. Furthermore, the open porosity, mercury intrusion porosimetry (MIP), x-ray diffraction (XRD) and thermogravimetric analysis (TGA) were examined to investigate the pore characteristics and phase composition of mortar. The results indicate that the resistance to chloride ingress and compressive strength of steam-cured mortar decline with the increase of fly ash incorporated, regardless of the subsequent curing condition. Compared to ZS, ZD and ZF lead to poor resistance to chloride penetration, while ZW and ZN show better performance. Interestingly, under different fly ash contents, the declining order of compressive strength remains ZS>ZW>ZN>ZD>ZF. When the fly ash content is blow 50%, the open porosity grows with increase of fly ash, regardless of the curing conditions are diverse. However, if the replacement amount of fly ash exceeds a certain high proportion (70%), the value of open porosity tends to decrease. Moreover, the main phase composition of the mortar hydration products is similar under different curing conditions, but the declining order of the C-S-H gels and ettringite content is ZS>ZD>ZF. The addition of fly ash could increase the amount of harmless pores at early age.

Identification of Retinol-binding Protein Produced by Caprine Endometrium during Periattachment Period of Early Pregnancy

  • Liu, K.H.;Huang, J.C.;Lin, J.H.
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.15 no.12
    • /
    • pp.1708-1713
    • /
    • 2002
  • Endometrial explants obtained from does between days 13 and 21 of pregnancy were cultured in a modified minimum essential medium in the presence of [$^35S$]methionine and [$^3H$]-leucine. Proteins synthesized and secreted into medium were analyzed by fluorography of two-dimensional polyacrylamide gel electrophoresis and fluorography. No marked qualitative changes in patterns of protein production by caprine endometrium between days 13-21 of pregnancy. At least 11 proteins showed consistently a clear spot or a grouping of spots with characteristic location on two-dimensional gels. A major low molecular weight protein consisted of two major isoforms (pI 5.3-6.0) of similar molecular mass (21 kDa). Limited N-terminal sequence analysis of these two isoforms showed that the protein had complete homology with bovine placental and plasma retinol-binding protein (RBP) over the first 20 amino acids. Through use of the antiserum raised against bovine placental RBP, immunoreactive RBP was detected in cultures conditioned by uterine explants prepared at days 13, 15 and 21 of pregnancy. In the present study, proteins synthesized and secreted by caprine endometrium during periattachment period of early pregnancy were characterized. The pregnant endometrium secreted a number of neutral-to-acidic proteins which constituted, in part, the histotroph. A vitamin A-transport protein, RBP, was identified in cultures conditioned by endometrium of days 13-21 of pregnancy. The uterine endometrium is the only source of retinol for embryonic tissues. The uterine RBP appears to transport retinol locally toward embryonic tissues. Secretion of RBP by caprine endometrium of days 13, 15 and 21 of pregnancy suggested that retinol played an important role in conceptus development during periattachment period of early pregnancy.

Analysis of Physical Properties for Various Compositions of Reusable LMG and LCV Micelle Gel

  • Kang, Jin Mook;Lee, Dong Han;Cho, Yu Ra;Hwang, Seon Bung;Ji, Young Hoon;Ahn, So Hyun;Keum, Ki Chang;Lee, Re Na;Cho, Sam Ju;Noh, Insup
    • Progress in Medical Physics
    • /
    • v.27 no.4
    • /
    • pp.175-179
    • /
    • 2016
  • In this study, we evaluated the reusable leuco malachite green (LMG) micelle gel properties dependent on various components of chemical concentration and compared with leuco crystal violet (LCV). The gels were delivered to 10, 20, 30, 40 and 50 Gy at 6 MV photon beam from linear accelerator and analyzed using spectrophotometry. We confirmed that the reusable LMG and LVC absorbance wavelength peak were made up at 630 nm and 600 nm respectively. The transparency of reusable LMG decreased with higher amount of trichloroacetic acid (TCAA) and lower reusable LMG dyes. 1 mM reusable LMG was the lowest transparency. The sensitivity was increased depending on lower trichloroacetic acid (TCAA) concentrations and the amount of suitable surfactant (Triton X-100), which was found to be 7 mM. However, we were not able to investigate sensitivity effects factor from reusable LMG dyes. The gel dosimeter containing 16 mM TCAA, 7 mM Triton X-100 gel dosimeter showed the highest sensitivity at $0.0021{\pm}0.0001cm^{-1}.Gy^{-1}$. The sensitivity of LCV was found to be higher than reusable LMG at $0.0037{\pm}0.0005cm^{-1}.Gy^{-1}$. The reusable LMG and LCV dose responses were shown to be $R^2=0.997$, $R^2=0.999$ respectively, as stable measurement results. Future research is necessary to improve dose sensitivity, dose rate dependency and gel fading with extensive chemical formulations.

Effect of Setting Temperatures and Time on the Gelation Properties (Suwari and Modori Phenomena) of Surimi from Mechanically Deboned Chicken Meat

  • Lee, Sung Ki;Min, Byung Jin
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.17 no.12
    • /
    • pp.1758-1763
    • /
    • 2004
  • This study was carried out to investigate gel forming and degradation properties (suwari and modori phenomena) of chicken surimi from mechanically deboned chicken meat (MDCM) at various setting temperatures and time. Chicken surimi was manufactured by a continuous process including chopping of MDCM, washing with 5% NaCl solution or pure water, standing, straining and centrifuging etc. Total process of washing for the MDCM from chopping to centrifuging was repeated over 3 cycles. Gel from prepared surimi were formed at $90^{\circ}C$ for 30 min after various setting treatments. The textural properties of gels were measured at the temperature ranges of low (10$^{\circ}C$), medium (25$^{\circ}C$ and $30^{\circ}C$) and high (45 to 70$^{\circ}C$). The compressive force (CF), hardness and fracturability of surimi gel at 10$^{\circ}C$ increased as setting time increased, and showed the highest value at 30 h of setting time. The CF and hardness of chicken surimi gel at 25$^{\circ}C$ and 30$^{\circ}C$ showed the highest values at 10 h of setting time. Most of gel strengths including CF, and texture profile analysis (TPA) values showed the highest levels in the range 47.5 to 52.5$^{\circ}C$ (p<0.05). The gel strength at 60$^{\circ}C$ increased slightly at 30 min, but then continued to decrease with longer setting times. There was no increase of gel strength at $70^{\circ}C$, but only a continuous decrease over setting time. In conclusion, suwari (gel setting) and modori (gel degradation) phenomena occur during the gel formation of surimi from MDCM. The temperature range in chicken surimi was 47.5 to 52.5$^{\circ}C$ for suwari and 60 to 70$^{\circ}C$ for modori.

Pilot Study of the Sensitivity and Specificity of the DNA Integrity Assay for Stool-based Detection of Colorectal Cancer in Malaysian Patients

  • Yehya, Ashwaq Hamid;Yusoff, Narazah Mohd;Khalid, Imran A.;Mahsin, Hakimah;Razali, Ruzzieatul Akma;Azlina, Fatimah;Mohammed, Kamil Sheikh;Ali, Syed A.
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.13 no.5
    • /
    • pp.1869-1872
    • /
    • 2012
  • Background: To assess the diagnostic potential of tumor-associated high molecular weight DNA in stool samples of 32 colorectal cancer (CRC) patients compared to 32 healthy Malaysian volunteers by means of polymerase chain reaction (PCR). Methods: Stool DNA was isolated and tumor-associated high molecular weight DNA (1.476 kb fragment including exons 6-9 of the p53 gene) was amplified using PCR and visualized on ethidium bromide-stained agarose gels. Results: Out of 32 CRC patients, 18 were positive for the presence of high molecular weight DNA as compared to none of the healthy individuals, resulting in an overall sensitivity of 56.3% with 100% specificity. Out of 32 patients, 23 had tumor on the left side and 9 on the right side, 16 and 2 being respectively positive. This showed that high molecular weight DNA was significantly (p = 0.022) more detectable in patients with left side tumor (69.6% vs 22.2%). Out of 32 patients, 22 had tumors larger than 1.0 cm, 18 of these (81.8%) being positive for long DNA as compared to not a single patient with tumor size smaller than 1.0 cm (p <0.001). Conclusion: We detected CRC-related high molecular weight p53 DNA in stool samples of CRC patients with an overall sensitivity of 56.3% with 100% specificity, with a strong tumor size dependence.