ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2013, Vol. 44 ›› Issue (11): 1832-1837.doi: 10.11843/j.issn.0366-6964.2013.11.019

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Studies on the Efficient Expression of African Swine Fever Virus p54 Protein and Its Antigenic Analysis in ELISA

GONG Zhen-hua1, WANG Li-ping1,2, ZANG Jing-shuai2, ZHANG Kang1,2, LIU Chun-ju1, WU Xiao-dong1, ZHANG Qi-di2, QIN Xiao-bing2, CHEN Lin-lin2, SHAN Hu2*, WANG Shu-shuang1, WANG Zhi-liang1*   

  1. (1. China Animal Health and Epidemiology Center, Qingdao 266032, China; 2.College of Animal Science and Technology, Qingdao Agricultural University, Qingdao 266109, China)
  • Received:2013-09-04 Online:2013-11-23 Published:2013-11-23

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Abstract:

In this study, p54 protein of African swine fever virus (ASFV) was prokaryotically expressed efficiently, and the expressed p54 protein was tested to be specific to ASFV in ELISA. A sequence-optimized P54 gene was synthesized with reference of P54 gene (E183L) sequence of ASFV Con09/Bzz020 strain, the synthesized ASFV P54 gene was cloned into pET-30c(+) and the recombinant plasmid pET-30c(+)-p54 was obtained. Nucleotide sequence analysis of pET-30c(+)-p54 showed that the synthesized sequence-optimized P54 gene was cloned successfully into pET-30c(+) with the right reading frame. The p54 fusion protein with approximately 20.7 kDa was expressed efficiently, solubly and stably, and easily to be purified with the purity at 900 μg·mL-1, the expressed p54 protein had the same amino acid sequence as the p54 protein of Con09/Bzz020, the expressed p54 protein was tested to be specific to anti-ASFV serum in Western-blot and had a P/N value of ASFV at 4.67 in ELISA. The results suggested that the expressed p54 protein with high yield was easy to be purified and was specific to anti-ASFV sera, and it could be used for diagnosis of ASFV by ELISA.

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