• Journal of Microbiology and Biotechnology
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• 제23권3호
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• pp.357-363
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• 2013

The identification of bacteriophage proteins on the surface of Lactobacillus rhamnosus Pen was performed by LC-MS/MS analysis. Among the identified proteins, we found a phage-derived major tail protein, two major head proteins, a portal protein, and a host specificity protein. Electron microscopy of a cell surface extract revealed the presence of phage particles in the analyzed samples. The partial sequence of genes encoding the major tail protein for all tested L. rhamnosus strains was determined with specific primers designed in this study. Next, RT-PCR analysis allowed detection of the expression of the major tail protein gene in L. rhamnosus strain Pen at all stages of bacterial growth. The transcription of genes encoding the major tail protein was also proved for other L. rhamnosus strains used in this study. The present work demonstrates the spontanous release of prophage-encoded particles by a commercial probiotic L. rhamnosus strain, which did not significantly affect the bacterial growth of the analyzed strain.

• International Journal of Oral Biology
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• 제30권1호
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• pp.31-37
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• 2005

Treponema maltophilum, a Group IV oral spirochete, is associated with periodontitis and endodontic infections. In this study we analyzed the functional role of the major surface protein of this organism (MspA) in human gingival fibroblasts (HGFs). The full-length gene encoding MspA was cloned and expressed in Escherichia coli by using the expression vector pQE-30. The recombinant protein (rMspA) was purified by affinity chromatography with nickel-nitrilotriacetic acid agarose and possible contamination of E. coli endotoxin in rMspA was removed by using polymyxin B-agarose. rMspA significantly induced the expression of pro inflammatory cytokines like IL-6 and IL-8 and intercellular adhesion molecule (ICAM)-1 in HGFs, when analyzed by reverse transcription-PCR, flow cytometry, and enzyme-linked immunosorbent assay. Our results indicate that MspA of T. maltophilum may play an important role in amplifying the local immune response by upregulating the expression of proinflammatory cytokines and ICAM-1.

• 약학회지
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• 제35권6호
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• pp.461-465
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• 1991

Silica-based gel filtration chromatography has been used to characterize molecular weight of proteins. However, the molecular weight measured by this method was distorted by protein-silica interactions like hydrophobic and electrostatic forces. Therefore, we characterized protein-silica interaction using two forms of phytochrome (124 kDa) having different hydrophobicity and surface charge. PH and ionic strength affected the retention time of phytochrome suggesting that electrostatic force is the major interaction between protein and silica surface.

• Parasites, Hosts and Diseases
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• 제45권1호
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• pp.59-63
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• 2007

Merozoite surface protein-1(MSP-1) and merozoite surface protein-2(MSP-2) were used to develop vaccines and to investigate the genetic diversity in Plasmodium falciparum malaria in Iran. Nested polymerase chain reaction amplification was used to determine polymorph isms of block 2 of the MSP-1 and the central domain of MSP-2 genes. A total of 67 microscopically positive P. falciparum infected individuals from a major endemic region, southeast Iran, were included in this trial. Nine alleles of MSP-1 and 11 alleles of MSP-2 were identified. The results showed that amplified product from these surface antigen genes varied in size and there was specific pattern for each isolate. Besides, regarding this pattern, 23 multiple infections with at least 2 alleles were observed. While the endemic regions of malaria in Iran is classified in low to moderate group, but extensive polymorphism was observed for each marker and the MSP-2 central repeat was the most diverse that could be considered in designing malaria vaccine.

• Parasites, Hosts and Diseases
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• 제37권4호
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• pp.277-283
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• 1999

To investigate the development of a subunit vaccine against theileriosis in cattle, the DNA fragments encoding piroplasm surface protein (p33) of Theileria sergenti of a Korean isolate were expressed in baculoviruses. The expressed p33 was characterized by indirect fluorescent antibody (IFA) and western blotting analysis. The expression of p33 was mainly detected on the surface of infected Sf21 cells by IFA. The immunoblotting analysis revealed the presence of a same molecular weight protein band of p33. The antigenicity of expressed polypeptide was further examined through the inoculation of a guinea pig. The sera of guinea pigs immunized with p33 expressed cell Iysate showed similar fluorescent antibody patterns and reacted with the same molecular weight protein of T. sergenti in immunoblotting analysis, thus indicating that this protein can be a promising candidate for a subunit vaccine in the future.

• Journal of Periodontal and Implant Science
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• 제46권1호
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• pp.2-9
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• 2016

Purpose: Porphyromonas gingivalis and Tannerella forsythia have been implicated as the major etiologic agents of periodontal disease. These two bacteria are frequently isolated together from the periodontal lesion, and it has been suggested that their interaction may increase each one's virulence potential. The purpose of this study was to identify proteins on the surface of these organisms that are involved in interbacterial binding. Methods: Biotin labeling of surface proteins of P. gingivalis and T. forsythia and liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis was performed to identify surface proteins involved in the coaggregating activity between P. gingivalis and T. forsythia. Results: It was found that three major T. forsythia proteins sized 161, 100, and 62 kDa were involved in binding to P. gingivalis, and P. gingivalis proteins sized 35, 32, and 26 kDa were involved in binding to T. forsythia cells. Conclusions: LC-MS/MS analysis identified one T. forsythia surface protein (TonB-linked outer membrane protein) involved in interbacterial binding to P. gingivalis. However, the nature of other T. forsythia and P. gingivalis surface proteins identified by biotin labeling could not be determined. Further analysis of these proteins will help elucidate the molecular mechanisms that mediate coaggregation between P. gingivalis and T. forsythia.

• 대한수의학회지
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• 제51권2호
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• pp.107-115
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• 2011

Theileria (T.) buffeli (formerly T. sergenti/T. orientalis) is the major hemo-protozoan distributed in the Far East Asian countries such as Korea, China and Japan. It is responsible for the clinical symptoms of anorexia, ateliosis, anemia, fever and icterus. It also causes abortion and sudden death under severe cases, resulting in economic losses for many livestock farms. The objective of this study was to analyze the genetic diversity of the major surface protein (Msp) gene in T. buffeli in Holstein in Korea, and we characterized the association of the diversification of the Msp gene and its relationship with the pathogenicity of Theileria. For this, complete blood counts and Theileria PCR sequence analysis were performed from 57 Holstein in Jeju Island. A total of 26 PCR positive Holstein (16 anemic and 10 non-anemic) were then randomly selected based on 18s rRNA sequence typing of the Theileria Msp gene. The DNA sequence of the T. buffeli Msp gene in Holstein showed 99.0%, 99.2%, 99.9%, 99.5%, 98.7%, 98.4% and 98.4% homology with T. sergenti, Theileria spp., T. sergenti, Theileria spp., Theileria spp., Theileria spp. and Theileria spp., respectively. The result showed a genetic variation of 57.7% (type I), 3.8% (type II), 15.4% (type III), 7.7% (type IV), 13.5% (type V) and 1.9% (type VI). Type I is the most frequent type in both anemic and non-anemic Holstein while type II was found in only non-anemic Holstein. This results of our study help confirm the diversity of Msp gene types and demonstrate that the gene type distribution of Msp genes varies among Theileria-infected Holstein in Jeju Island.

• Food Science and Biotechnology
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• 제17권2호
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• pp.324-329
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• 2008

The effect of ultra-high pressure homogenization on the emulsifying properties of whey protein was investigated in a model emulsion made with whey protein isolate and soya oil under various pH. The emulsifying properties, the average diameter of the oil droplets ($d_{vs}$), and the protein load, were measured for each emulsion produced at different homogenization pressures (50 to 200 MPa) and pH values (4.6 to 8.0). According to the results of variance analysis and response surface, the pH had more influence on oil droplet size and protein load than homogenization pressure. The model equations, which were obtained by response surface analysis, show that pH and homogenization pressure had the major effect on oil droplet size and protein load. Higher homogenization pressure decreased the average droplet size and the protein load. Homogenization at high pressure, as opposed to low pressure, causes no overprocessing, but the effect was pH-dependent. The average diameter of the oil droplets increased slightly by decreasing the pH from 8.0 to 6.5 and then increased dramatically toward the isoelectric point of whey protein (i.e., at pH 4.6). Moreover associated droplets were found at acidic pH and their size was increased at high temperature.

• Archives of Pharmacal Research
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• 제21권5호
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• pp.521-526
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• 1998

The physicochmical properties of recombinant hepatitis B surface antigen (r-HBsAg), which was expressed in C127 mammalian cell were studied. Using roller bottle culture in DMEM supplemented with fetal bovine serum, 10-15 mg/L of r-HBsAg was produced with about 31% of purification yield. The purity of r-HBsAg by HPLC was 99.8% and electron microscopic examination showed homogeneous spherical particle with 22 nm in diameter, a morphological characteristic of HBsAg. The density of r-HBsAg by CsCI density gradient method was 1.19g/ml and the isoelectric point by Mono $P^{TM}$ HR 5/20 column was 4.6. The analysis of subunit protein pattern using SDS-PAGE followed by scanning densitometry gave 81.3% of S protein and 18.7% of pre-S protein. fluorophore-assisted-carbohydrate-electrophoresis analysis showed the relative amount of carbohydrate to protein was 1.7% and it smajr component was N-acetyl glucosamine, which was about 39% of total carbohydrate. The relative amount of lipid to protein determined by vanillin phosphoric acid method was 32.5% and its major component was phospholipid, which was about 70% of total lipid. The physicochemical properties of C127 mammalian cell-derved r-HBsAg are similar to those of p-HBsAg, suggesting that the r-HBsAg can be used in developing a new preventive vaccine against hepatitis B.

• Parasites, Hosts and Diseases
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• 제26권3호
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• pp.155-162
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• 1988

Toxoplasmn gondii이 강독주인 RH주와 조직내 cyst 형성주인 Fukaya주의 세포막 단배 성분을 SDS 존재하에 서 전기영동하여 분석하였다. 먼저 RH tachyzoite와 Fukaya의 cyst를 각각 마우스의 복강액과 뇌조직으로부터 분리하였는데, 불연속 Percoll density-gradient서 원심분리하여 tachygoite는 50 U와 605 Percoll용액 경계면에서, cyst는 40%와 50%의 경계면 및 50%와 60 % 경계면에서 얻었으며, cyst는 저장액으로 처리하여 bradyzoite를 얻었다. Lactoperoxidase를 촉매로 세포막에 방사성 요오드를 표지시킨 후 자가방사표지그림을 얻었을 때, bradyzoite 는 15 KDa와 14 KDa의 분자량을 가진 단백질이 주요 단백질로 나타났으며, tachyzoite에서는 30 KDa 단백질이 주요 단백질로 나타났다. 또, 당단백질의 존재를 파악하기 위해서 lectin blotting을 시행하였는데, concanavalin A는 bradyzoite에서 200K∼50KDa의 여러 단백질을, .그리고 tachyzoite에서는 52KDa 단백질을 주로 하는 33K∼20 KDa단백질을 검출하였으며, phytohemagglutinin은 두·유형에서 아무런 단백질도 검출하지 못하였다. 한편, 이들을 효소면역이적법으로 항 Fukfya항체와 항 RH항체로 반응시켰을 때, 많은 교차 반응을 보였으나, bradyzoite에서는 15 KDa 단백질이, 그리고 tachyzoite에서는 52 KDa, 30 KDa 및 25 KDa 단백 짙이 각각 유형 특이 항원 단백으로 나타났다. 위의 결과들로, bradyzoite에서는 15 KDa 단백질이 당단백질은 아니지만 특이 항원성을 갖는 주요 백으로 나타났으며, tachyzoite에서는 지금까지 주요 세포막 단백으로. 알려진 P3O외에 당단백질이며 성을 갖는 세포막 단백으로 SaKDa 단백 (gps2)을 확인할 수 있었다.