猪圆环病毒2型病毒样颗粒的纯化及鉴定

doi:10.16431/j.cnki.1671-7236.2019.06.027

Abstract

Abstract:

To obtain and analyze highly purified porcine circovirus virus type 2 (PCV2) virus-like particles(VLPs),the PCV2 recombinant Cap protein (PCV2 rCap) was expressed by Escherichia coli expression system,and was purified by the means of ammonium sulfate precipitation,gel filtration chromatography,ion exchange chromatography and CsCl density gradient centrifugation.The immunoreactivity of purified VLPs were identified by Western blotting,the diameter and morphology of purified VLPs were observed by transmission electron microscopy (TEM).Its components were detected and analyzed by high performance liquid size exclusion chromatography (HPSEC).After purification,the PCV2 rCap protein was detected by SDS-PAGE,the target protein band of 28 ku was detected,and its purity was 97.53% by gray scale scanning.The results of Western blotting showed that the specific protein strips had obvious immunoreactivity.The TEM detected the regular spherical particles with the particle size of 17.35 to 19.24 nm,indicating that the obtained PCV2 rCap protein could express itself in the bacteria.They were assembled into VLPs and deagglomerated without being destroyed during the purification process,and remained in a natural state;In addition,HPSEC detected that the proportion of normally assembled VLPs was 92.67%.The results of this study provided a reference for further study of the spatial structure of PCV2 VLPs and their immune protection mechanisms.

Key words: porcine circovirus type 2(PCV2); virus-like particles(VLPs); purification; identification; gel filtration chromatography; ion exchange chromatography

CLC Number:

S852.65

WANG Xi, LI Guopan, CHEN Qingqing, XU Baojuan, RONG Jun. Purification and Identification of Porcine Circovirus Type 2 Virus-like Particles[J]. , 2019, 46(6): 1792-1800.

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Purification and Identification of Porcine Circovirus Type 2 Virus-like Particles

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