[Prokaryotic expression and identification of S-dsRNA gene from Cryptosporidium parvum virus].

Yu mei Diao, Peng tao Gong, Wei Li, Li bo Su, Xiangsheng Huang, Jian hua Li, Xi chen Zhang

Research output: Contribution to journalArticle

Abstract

To clone and express S-dsRNA gene of Cryptosporidium parvum virus, and investigate the reactogenicity of the recombinant. Total RNA was extracted from Cryptosporidium parvum and S-dsRNA gene was amplified by RT-PCR. The PCR product was cloned into pET-28a(+) expression vector. The recombinant plasmid pET-28a(+)-S was transformed into E. coli BL21 (DE3) and induced with IPTG. The expression situation of recombinant protein was analyzed by SDS-PAGE. Its reactogenicity was examined by Western blotting analysis. pET-28a (+)-S was identified by PCR and double endonuclease digestion. SDS-PAGE result showed that the recombinant protein (M, 37,000) was expressed in the form of inclusion body. High level expression of recombinant protein was found at 1 mmol/L IPTG condition after incubation at 37 degrees C for 4 h and reached up to 72.6% of the total protein. The protein was recognized by the antisera from mice immunized with antigens from Cryptosporidium parvum oocysts. The S-dsRNA gene of Cryptosporidium parvum virus has been expressed with adequate reactogenicity.

Original languageEnglish (US)
Pages (from-to)29-32
Number of pages4
JournalZhongguo ji sheng chong xue yu ji sheng chong bing za zhi = Chinese journal of parasitology & parasitic diseases
Volume29
Issue number1
StatePublished - Feb 28 2011
Externally publishedYes

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Cryptosporidium parvum
Recombinant Proteins
Viruses
Isopropyl Thiogalactoside
Polymerase Chain Reaction
Genes
Polyacrylamide Gel Electrophoresis
Oocysts
Endonucleases
Inclusion Bodies
Immune Sera
Digestion
Proteins
Plasmids
Clone Cells
Western Blotting
RNA
Escherichia coli
Antigens

ASJC Scopus subject areas

  • Medicine(all)

Cite this

[Prokaryotic expression and identification of S-dsRNA gene from Cryptosporidium parvum virus]. / Diao, Yu mei; Gong, Peng tao; Li, Wei; Su, Li bo; Huang, Xiangsheng; Li, Jian hua; Zhang, Xi chen.

In: Zhongguo ji sheng chong xue yu ji sheng chong bing za zhi = Chinese journal of parasitology & parasitic diseases, Vol. 29, No. 1, 28.02.2011, p. 29-32.

Research output: Contribution to journalArticle

Diao, Yu mei ; Gong, Peng tao ; Li, Wei ; Su, Li bo ; Huang, Xiangsheng ; Li, Jian hua ; Zhang, Xi chen. / [Prokaryotic expression and identification of S-dsRNA gene from Cryptosporidium parvum virus]. In: Zhongguo ji sheng chong xue yu ji sheng chong bing za zhi = Chinese journal of parasitology & parasitic diseases. 2011 ; Vol. 29, No. 1. pp. 29-32.
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