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)
文献类型： 外文期刊
作者： Chen, H 1 ; Lu, JH 2 ; Liang, WQ 2 ; Huang, YH 2 ; Zhang, WJ 2 ; Zhang, DB 2 ;
作者机构： 1.Shanghai Jiao Tong Univ, Coll Life Sci & Biotechnol, Shanghai 200030, Peoples R China
2.Shanghai Jiao Tong Univ, Coll Life Sci & Biotechnol, Shanghai 200030, Peoples R China; Shanghai Univ, Sch Life Sci, Shanghai 200436, Peoples R China; Shanghai Inst Appl Phys, Shanghai 201800, Peoples R China; Shanghai Acad Agr Sci, Agro Biotech Res Ctr, Shanghai 201106, Peoples R China; Nanjing Univ, Dept Biol Sci & Technol, Nanjing 210093, Peoples R China
关键词： rHBcAg purification;Saccharomyces cerevisiae;rHBcAg (core) particles (capsids);transmission electron microscopy;atomic force microscopy;dimorphism
期刊名称：MICRON （ 影响因子：2.251； 五年影响因子：1.969 ）
ISSN： 0968-4328
年卷期： 2004 年 35 卷 5 期
页码：
收录情况： SCI
摘要： Hepatitis 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.
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