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곤충세포에서 인유두종 바이러스 18형의 바이러스 유사 입자 생산
강병태, 박순희 김천대학교 1999 김천대학교 논문집 Vol.20 No.-
To express the Capsid protein feom human Paillomavirus(HPV) type 18 in insect cells, we have cloned L1 and L2 open reading frame(ORF). into baculovirus expression vector. pAcUw 31. contained dual promotor. The recombinant plasmid, pAc/18 L1-L2 was transfected into Sf-21 and the high titier recombinant baculovirus was then taken through plaque pruification. From insect cells infected with the recombinant virus, the capsid proteins were produced and confirmed by western blot analysis. These proteins were detedced the mobility of 63Kd by HPV16L1 monoclonal antibody for 18L1 and of 70Kd by TrpE-HPV 18L2 polyclonal antibody for 18L2, respectively. But the expression level of capsid protein in dual premoter-based vector were much lower than that of those proteins in single promotor-based vector. The 3-day-infected cellular lysates from this recombinant virus was purified by cesium chloride gradient centrifugation. Viruslike particles(VLPs) were found to have a buoyant density of 1.29 g/ml and measured a diameter of 45-55 nm, which corresponds to that of native papillomavirus virions. The micrographs also showed that the another portion of VLPs was prepared for the abnormal shape as elliptical form or capsomers. It assumed that VLPs may be on progress the assembly or be made with the unregular assembly.
Park, Sue Nie,Lee, Kyung Ae,Cho, Young Sik,Choe, Yong Kyung,Yoon, Do Young,Kwon, Dur Han,Kang, Jeong Woo,Cho, Cheong Weon,Cho, Min Chul,Shim, Jung Hyun 생화학분자생물학회 2002 BMB Reports Vol.34 No.1
The human papillomavirus E7 pmtein can form a specific complex with a retinoblastoma tumor suppressor gene product (p105-Rb) that results in the release of the E2F transcription factor, which is critical for the growth-deregulation and transforming properties of the viral E7 oncoprotein. In an attempt to apply interaction between the E7 oncoprotein and a target cellular protein Rb for an in vitro screening system for drugs against human papillomavirus infection, we primarily investigated the E7Rb binding through a pull down assay and enzyme-linked immunosorbent assay. The pull down assay showed that both glutathione S-transferase-tagged E7 and His-tagged E7 immobilized on resins specifically produced complexes with bacterially expressed Rb in a dose-dependent manner, as determined by immunoblot analyses. This result coincided with that of an enzyme-linked immunosorbent assay, which is a useful system for the mass screening of potential drugs. Taken together, this screening system (based on the interaction between E7 and Rb) can be a promising system in the development of drugs against cervical cancers caused by human papillomavirus infection.
( Sue Nie Park ) 한국동물실험대체법학회 2007 한국동물실험대체법학회 학술대회집 Vol.2007 No.1
Recently, the understanding of the mechanism of genotoxicity and / or the prediction of carcinogenicity of chemicals have been attempted by toxicogenomics. However, the procurement of enough amount data set for prediction analysis is usually not so easy for the costs. Therefore, it would be useful to improve the prediction power by applying proper analytical methods on the data set obtained for small number of chemicals. Thus, in this study, we attempted to apply and compare the classification approaches such as k-NN(k-nearest Neithborhood), SVM(Support Vector Machine), PLS(Partial Least Square regression), PCR(Principle Component regression), and ridge regression discriminant classification and compared to ensemble method with bootstrap resampling. Four chemicals each representing genotoxic carcinogen(glycidol), genotoxic noncarcinogen(8-hydroxyquinoline), nongenotoxic carcinogens(o-nitrotoluene), and nongenotoxic noncarcinogen(1,2-dichlorobenzene) were selected to evaluate the classification methods using microarray chip data. Each minimal error rate was 0.13, 0.0, 0.0, 0.0, and 0.0 for training step while 0.8, 0.4, 0.8, 0.8, and 0.8 was obtained in test data, respectively. The application of LogitBoost resulted in minimum error of 0.13 in training data and 0.3 in test data with improvement of prediction power. Our study may offer the more effective method in evaluating toxicity of chemicals related to genotoxicity and / or predicition of carcinogenecity.
Ryu, Sung-Weon,Park, Sang-Chan,Bang, Mun-Nam,Han, Sung-Sik,Park, Young-Kil,Park, Sue-Nie,Shim, Young-Soo,Kan, Seong-Man,Bail, Gill-Han The Korean Society for Microbiology 2004 Journal of Bacteriology and Virology Vol.34 No.2
Despite recent economic prosperity, Korea still has high prevalence of tuberculosis. Molecular biologic characterization of Korean Mycobacterium tuberculosis strains might provide a deeper understanding of the forces contributing to the spread of tuberculosis in Korea. Therefore, we analyzed the cell lysate proteome of a representative Korean Mycobacterium tuberculosis isolate (K01) in comparison with laboratory reference strains H37Rv and H37Ra. Seven spots were strongly expressed only in K01 strain compared with M. tuberculosis H37Rv and H37Ra. Through continuous MALDI-MS analysis, these spots were identified as hypothetical protein Rv3849, secreted immunogenic protein Mpt64, Acetyl/propionyl-CoA Carbpxylase (AceD1), alkyl hydroperoxide reductase C (AhpC), N-acetylmuramyl-L-alanine amidase, a putative UDP glucose epimerase, and a transposase. A deeper study of these proteins may provide a clue in the development of effective new anti-tuberculosis vaccines against Korean M. tuberculosis isolates.
Ryoo, Sung-Weon,Park, Young-Kil,Park, Sue-Nie,Shim, Young-Soo,Liew, Hyun-Jeong,Kang, Seong-Man,Bai, Gill-Han The Microbiological Society of Korea 2007 The journal of microbiology Vol.45 No.3
In Korea, the Mycobacterium tuberculosis K-strain is the most prevalent clinical isolates and belongs to the Beijing family. In this study, we conducted comparative porteomics of expressed proteins of clinical isolates of the K-strain with H37Rv, H37Ra as well as the vaccine strain of Mycobacterium bovis BCG following phagocytosis by the human monocytic cell line U-937. Proteins were analyzed by 2-D PAGE and MALDI-TOF-MS. Two proteins, Mb1363 (probable glycogen phosphorylase GlgP) and MT2656 (Haloalkane dehalogenase LinB) were most abundant after phagocytosis of M. tuberculosis K-strain. This approach provides a method to determine specific proteins that may have critical roles in tuberculosis pathogenesis.