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Purification of the recombinant hepatitis B virus core antigen (rHBcAg) produced in the yeast Saccharomyces cerevisiae and comparative observation of its particles by transmission electron microscopy (TEM) and atomic force microscopy (AFM)
Chen, H; Lu, JH(陆军洪); Liang, WQ; Huang, YH; Zhang, WJ; Zhang, DB; Chen, H (reprint author), Shanghai Jiao Tong Univ, Coll Life Sci & Biotechnol, Shanghai 200030, Peoples R China
2004
Source PublicationMICRON
ISSN0968-4328
Volume35Issue:5Pages:311
AbstractHepatitis B virus core antigen (HBcAg) gene (C gene) was expressed in Saccharomyces cerevisiae and the products (rHBcAg or core particles) were purified from a crude lysate of the yeast by three steps: Sephrose CL-4B chromatography, Sucrose step-gradient ultracentrifugation and CsCl-isopycnic ultracentrifugation. It has been observed that HBcAg was synthesized in yeast cells as a particle consisting of polypeptides with a molecular weight of 21.5 kDa (p21.5). Results of ELISA test and density analysis of CsCl-isopycnic ultracentrifugation indicated that the purified products (rHBcAg particles) with HBcAg antigenicity mainly located at the densities of 1.27 and 1.40 g ml(-1), respectively. Observation and analysis of the purified rHBcAg products by TEM indicated that rHBcAg peptides could mainly self-assemble into two size classes of core particles. The larger particles were similar to 30.1 nm and the smaller were similar to 21.5 nm in mean diameter. Further observation and analysis of the same rHBcAg (core) particles by AFM also indicated that rHBcAg (core) particles were similar to the native HBcAg (core) particles from infected human hepatocytes and mainly composed of two size classes of particles core. The larger particles were similar to 3 1.3 nm and the smaller were similar to 22.5 mn in mean diameter which was similar to the results obtained by TEM. All results from both TEM and AFM suggested that core particles (capsids) produced in S. cerevisiae possessed dimorphism. (C) 2004 Elsevier Ltd. All rights reserved.
KeywordRhbcag Purification Saccharomyces Cerevisiae Rhbcag (Core) Particles (Capsids) Transmission Electron Microscopy Atomic Force Microscopy Dimorphism
Subject AreaChemistry ; Physics
Indexed BySCI
Language英语
Funding Project应物所项目组
Document Type期刊论文
Identifierhttp://ir.sinap.ac.cn/handle/331007/8172
Collection中科院上海应用物理研究所2004-2010年
Corresponding AuthorChen, H (reprint author), Shanghai Jiao Tong Univ, Coll Life Sci & Biotechnol, Shanghai 200030, Peoples R China
Recommended Citation
GB/T 7714
Chen, H,Lu, JH,Liang, WQ,et al. Purification of the recombinant hepatitis B virus core antigen (rHBcAg) produced in the yeast Saccharomyces cerevisiae and comparative observation of its particles by transmission electron microscopy (TEM) and atomic force microscopy (AFM)[J]. MICRON,2004,35(5):311.
APA Chen, H.,Lu, JH.,Liang, WQ.,Huang, YH.,Zhang, WJ.,...&Chen, H .(2004).Purification of the recombinant hepatitis B virus core antigen (rHBcAg) produced in the yeast Saccharomyces cerevisiae and comparative observation of its particles by transmission electron microscopy (TEM) and atomic force microscopy (AFM).MICRON,35(5),311.
MLA Chen, H,et al."Purification of the recombinant hepatitis B virus core antigen (rHBcAg) produced in the yeast Saccharomyces cerevisiae and comparative observation of its particles by transmission electron microscopy (TEM) and atomic force microscopy (AFM)".MICRON 35.5(2004):311.
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