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Updated: Aug 16, 2026
Generation and Assembly of Virus-Specific Nucleocapsids of the Respiratory Syncytial Virus
Published on: July 27, 2021
[Purification and structural analysis of the nucleocapsid of PRRS virus expressed from Escherichia coli]
Jin-Hai Huang1, Han-Chun Yang, Xin Guo
1Key Laboratory of Preventive Veterinary Medicine of Ministry of Agriculture, College of Veterinary Medicine, China Agirictural University, Beijing 100094, China.
Abstract:
The DNA fragment encoding the nucleocapsid protein (N) of PRRSV BJ4 strain were cloned into the BamH I / EcoR I sites of pET28a vector to construct the expression plasmid pET28-N by designing special primers. The soluble protein (P28-N) were obtained by introducing the expression plasmid into E. coli BL21 (DE3) host cell, and the amount of recombinant protein reached to 28% of the total mass of bacterial protein. PET28-N were purified by nickel-affinity column of Proband resin. The circular dichroism (CD) analysis showed that the purified PET28-N shared a significant (26%) alpha-helical structure, beta-sheet (23.7%), beta-turn (19.8%), and random coil (30.3%), respectively. Finally,the secondary structure of N protein of PRRSV was deduced.
