• 제목/요약/키워드: SDS-PAGE gel electrophoresis

검색결과 340건 처리시간 0.025초

Extracellular synthesis of silver nanoparticle by Pseudomonas hibiscicola - Mechanistic approach

  • Punjabi, Kapil;Mehta, Shraddha;Yedurkar, Snehal;Jain, Rajesh;Mukherjee, Sandeepan;Kale, Avinash;Deshpande, Sunita
    • Advances in nano research
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    • 제6권1호
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    • pp.81-92
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    • 2018
  • Biosynthesis of nanoparticles has acquired particular attention due to its economic feasibility, low toxicity and simplicity of the process. Extracellular synthesis of nanoparticles by bacteria and fungi has been stated to be brought about by enzymes and other reducing agents that may be secreted in the culture medium. The present study was carried out to determine the underlying mechanisms of extracellular silver nanoparticle synthesis by Pseudomonas hibiscicola isolated from the effluent of an electroplating industry in Mumbai. Synthesized nanoparticles were characterized by spectroscopy and electron microscopic techniques. Protein profiling studies were done using Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (1D-SDS PAGE) and subjected to identification by Mass Spectrometry. Characterization studies revealed synthesis of 50 nm nanoparticles of well-defined morphology. Total protein content and SDS PAGE analysis revealed a reduction of total protein content in test (nanoparticles solution) samples when compared to controls (broth supernatant). 45.45% of the proteins involved in the process of nanoparticle synthesis were identified to be oxidoreductases and are thought to be involved in either reduction of metal ions or capping of synthesized nanoparticles.

산천어의 바이러스성 질병에 관한 연구-II -산천어 치어에서 1HNV 분리- (Studies on Viral Disease of masu salmon, Oncorhynchus masou-II Isolation of infectious hematopoietic necrosis virus form masu salmon fry)

  • 손상규;박명애;박정우
    • 한국어병학회지
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    • 제6권2호
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    • pp.87-92
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    • 1993
  • 1. 1990년 2월 강원도 삼척군에 소재한 송어양식장에서 전염성 조혈기 괴사증 증상으로 산천어 자치어가 대량폐사하였다. 2. 폐사한 산천어 치어로부터 세포배양법에 의해 전염성 조혈기 괴사증 바이러스가 분리되었다. 3. 분리된 바이러스는 구조단백의 크기와 항원성에 있어 미국에서 분리된 RB-76 바이러스주(electro-pherotype 1)와 유사하였다.

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Cloning and Expression of Mycobacterium bovis Secreted Protein MPB83 in Escherichia coli

  • Xiu-Yun, Jiang;Wang, Chun-Feng;Wang, Chun-Fang;Zhang, Peng-Ju;He, Zhao-Yang
    • BMB Reports
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    • 제39권1호
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    • pp.22-25
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    • 2006
  • The gene encoding MPB83 from Mycobacterium bovis Vallee111 chromosomal DNA was amplified by using polymerase chain reaction (PCR) technique, and the PCR product was approximately 600bp DNA segment. Using T-A cloning technique, the PCR product was cloned into pGEM-T vector and the cloning plasmid pGEM-T-83 was constructed successfully. pGEM-T-83 and pET28a(+) were digested by BamHI and EcoRI double enzymes. The purified MPB83 gene was subcloned into the expression vector pET28a(+), and the prokaryotic expression vector pET28a-83 was constructed. Plasmid containing pET28a-83 was transformed into competence Escherichia coli BL21 (DE3). The bacterium was induced by isopropyl-$\beta$-D-thiogalactopyranoside (IPTG) and its lysates were loaded directly onto sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE), approximately 26 kDa exogenous protein was observed on the SDS-PAGE. The protein was analyzed using Western-blotting. The results indicated that the protein was of antigenic activity of M. bovis. The results were expected to lay foundation for further studies on the subunit vaccine and DNA vaccine of MPB83 gene in their prevention against bovine tuberculosis.

Cloning and Expression of Human KCNE1 Gene

  • ;;;유민
    • 대한의생명과학회지
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    • 제16권4호
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    • pp.299-305
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    • 2010
  • KCNE1 is the causal gene of long QT syndrome. KCNE1 gene is located in chromosome 21. In compliance with this KCNE1 gene the proteins come out. KCNE1 is responsible for $K^+$ channel which maintains the normal function of the heart muscle for contraction. Affected individuals manifest prolongation of the QT interval on electrocardiongrams, a sign of abnormal cardiac repolarization. The clinical features of LQT result from episodic cardiac arrhythmias, such as torsade de pointes and ventricular fibrllation. Blood DNA was isolated and kept in $4^{\circ}C$ refrigerator. The KCNE1 gene was amplified by PCR method and about 414 bp band was identified by agarose gel electrophoresis. PCR products were inserted into pGEX-4T-1 vector in order to express KCNE1 protein after treatment with IPTG SDS-PAGE was carried out and the protein band which was about 47 kDa was clearly odserved. Results of this study would contribute to the detailed understanding of KCNE1 protein function and to designing better treatment of Long QT symdrome.

SOLVING BIOANALYTICAL PROBLEMS BY THE METHOD OF MATRIX-ASSISTED LASER DESORPTION IONIZATION MASS SPECTROMETRY (MALDI-MS)

  • Zhao, Shankai;Zhong, Feng;Zhu, Zhihua
    • 분석과학
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    • 제8권4호
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    • pp.865-868
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    • 1995
  • The method of matrix-assisted laser desorption ionization mass spectrometry has been used to solve some bioanalytical problems, which is difficult to analyse by general methods. For the selection of proper laser wavelength and matrices, eight matriees was used with laser wavelength of 226 and 355nm. The result shows that with wavelength of 355nm better results could be obtained with most of the matrices. The molecular weight of eytochrome C, which was seperated by gel electrophoresis and electro-blotted onto NC membrane is determined by MALDI. The accuracy is better than 0.1%, which is much higher than that of SDS-PAGE. Protein mixture extracted from crude peanut oil is directly determined by MALDI. The molecuiar weight of its three components are determined, and the result also demonstrated that these proteins are in free manner. As proteins arc in 2S bond, with the traditional method, SDS-PAGE, it is not able to decide whether protein exists in combination mode or in free manner. In the technique of two phase aquesous solution, which is used for separating biomaterials, water soluble polymers stained with dyes are used in this technique. By the use of MALDI the number or the dye molecules react with the polymer PEG molecule are determined, and that is difficult to determined by other methods.

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Characterization of Edible Film Fabricated with Channel Catfish Ictalurus punctatus Gelatin by Cross-Linking with Transglutaminase

  • Oh, Jun-Hyun
    • Fisheries and Aquatic Sciences
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    • 제15권1호
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    • pp.9-14
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    • 2012
  • The objectives of this research were to improve the film-forming properties of Channel Catfish Ictalurus punctatus skin gelatin (CSG) by cross-linking with transglutaminase (TG), determine and optimize the TG reaction time, and characterize the mechanical and barrier properties of CSG edible film. Cross-linking of CSG was performed by TG for 0, 10, 20, 30, and 40 min at $50^{\circ}C$, and the reaction was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The color and mechanical and barrier properties of edible films fabricated with CSG cross-linked with TG were characterized. Gelatin yields from the extraction ranged from 18.2% to 23.3%. SDS-PAGE exhibited dark bands at 120 and 250 kDa, indicating successful TG-mediated cross-linking. The color of CSG film was not affected by TG cross-linking. The tensile strength of CSG films cross-linked with TG decreased from 42.59 to 21.73 MPa and the percent elongation increased from 42.92% to 76.96% as reaction times increased from 0 to 40 min. There was no significant difference in water vapor permeability of CSG films.

정상치은열구액과 치주질환시 치은열구액의 단백질 조성비교에 관한 연구 (THE PROTEIN COMPOSITION OF GINGIVAL CREVICULAR FLUID SAMPLED FROM NORMAL SUBJECTS AND PATIENTS WITH PERIODONTAL DISEASE)

  • 김수아;이진용;김형섭
    • Journal of Periodontal and Implant Science
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    • 제23권3호
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    • pp.391-398
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    • 1993
  • Gingival crevicular fluid (GCF) is a promising source for markers of destructive periodontal disease activity. This study was undertaken to evaluate the protein composition of GCF in varying stages of the gingival inflammatory response. GCF sampled from 26 people with clinically healthy gingiva and 18 people with periodontitis were examined via sodium dodecyl sulphate polyacrylamide gel electrophoresis(SDS/PAGE). The result were as follows. 1. Total amount of GCF protein of diseased group significantly different from that of normal group. But difference in protein concentration was not that significant. 2. In analyzing GCF with SDS/PAGE, it was suggested that albumin is used as indicator plasma protein leakage because of heavily staining bond of albumin in patients with periodontal disease. 3. In diseased group, overall bonds of protein and bands of high molecular weight protein were heavily stained. It was proved useful information on high molecular plasma protein leakage with increasing vascular permeability due to inflammation.

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정제된 먹물버섯(Coprinus comatus) laccase에 의한 염료 탈색 (Decolorization of dyes by a purified laccase from Coprinus comatus)

  • 김수연;최지영;최형태
    • 미생물학회지
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    • 제53권2호
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    • pp.79-82
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    • 2017
  • 먹물버섯 Coprinus comatus를 yeast extract peptone dextrose 액체배지에서 배양하고 laccase의 생성 분비를 확인하였다. 이 때 분비된 laccase를 이온교환수지 방법과 전기영동 분획수집방법으로 순수하게 정제하였다. 정제된 효소단백질의 분자량은 SDS-PAGE로 분석하였을 때 약 67 kDa으로 나타났고 최적산도는 pH 4.3, 최적온도는 $25^{\circ}C$로 확인되었다. 기질 o-tolidine에 대한 Km 값은 0.45 mM, Vmax 값은 0.0132 OD/min/unit로 나타났다. 정제된 laccase를 사용하여 염료 RBBR과 Poly R-478의 탈색을 분석한 결과 배양 12시간 후에 RBBR은 49.3% 탈색되었고 poly R-478은 41.6% 탈색되었다.

Quality properties of whole milk powder on chicken breast emulsion-type sausage

  • Kang, Kyu-Min;Lee, Sol-Hee;Kim, Hack-Youn
    • Journal of Animal Science and Technology
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    • 제63권2호
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    • pp.405-416
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    • 2021
  • The aim of the study was to determine the effect of whole milk powder (WMP) as heterologous proteins on chicken breast emulsion-type sausages. The quality properties of WMP on such chicken breast emulsion-type sausages were investigated by measuring the proximate composition, pH, color, cooking yield, protein solubility, and by applying other methods, such as texture profile analysis (TPA), microphotograph, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and electronic nose. The crude fat, protein, and ash contents of 15% WMP samples were significantly higher than the control samples (p < 0.05). The redness of the cooked samples significantly increased with an increase in the WMP contents (p < 0.05). The cooking yield of WMP treated samples was significantly higher than the control sample (p < 0.05). Additionally, the hardness, gumminess, and chewiness of WMP treated samples were significantly higher than the control sample (p < 0.05). The sarcoplasmic and myofibrillar proteins of samples containing 15% WMP were significantly higher than the control samples (p < 0.05). The result of SDS-PAGE showed that the C protein, sarcoplasmic protein, actin, and tropomyosin increased with an increase in the WMP contents. The principal component analysis plot of WMP-treated samples was clearly different from that of the control samples. Based on these results, it was predicted that WMP could be useful as heterologous protein on emulsion-type sausage.

Effects of Using Soybean Protein Emulsion as a Meat Substitute for Chicken Breast on Physicochemical Properties of Vienna Sausage

  • Kang, Kyu-Min;Lee, Sol-Hee;Kim, Hack-Youn
    • 한국축산식품학회지
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    • 제42권1호
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    • pp.73-83
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    • 2022
  • The aim of this study is to determine the effects of using emulsion manufactured with soybeans (ES) to substitute chicken breast in Vienna sausages. Four types of Vienna sausages (S1: 10% ES and 50% chicken, S2: 20% ES and 40% chicken, S3: 30% ES and 30% chicken, and S4: 40% ES and 20% chicken) for this study were made. The pH, color, proximate composition, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), microphotographs, cooking yields, and texture profile analysis of sausages were examined. The pH value of uncooked and cooked sausages increased significantly with increasing ES content (p<0.05). The crude protein contents of S2, S3, and S4 were significantly higher than that of the control (p<0.05). Furthermore, the SDS-PAGE results showed that α-conglycinin, β-conglycinin, and the acidic subunit of glycinin all increased with increasing ES content. Microphotographs revealed that increasing the ES content decreased the size of fat globules. The cooking yields of samples increased significantly with increasing ES content (p<0.05). The hardness values of ES treated samples were significantly lower than that of the control (p<0.05). Therefore, 30% substitute of chicken breast with ES can improve the quality and structure of Vienna sausage, without inducing critical defects.