Construction of an artificial recombinant bicistronic plasmid DNA vaccine against porcine rotavirus

Onderstepoort Journal of Veterinary Research

Qin Zhao¹  Xintian Wen²  Xiaoping Ma²  Yong Huang²  Xiaobo Huang²  Tingting Cui²  Sanjie Cao²  Jun Xiong²  Zhongkai Hu²  Yongzhi Wang³ 

[1] College of Food Science, Sichuan Agricultural University;College of Veterinary Medicine and Institute of Preventive Veterinary Medicine, Sichuan Agricultural University;Sichuan Rural Science and Technology Development Centre, Chengdu;

关键词: DNA vaccine;    immune response;    NSP4 gene;    porcine rotavirus;    Salmonella typhimurium χ4550 strain;    VP7 gene;   

DOI  :  

来源: DOAJ

【 摘 要 】

The attenuatedSalmonella typhimuriumχ4550 strain was used to harbour a reconstructedbicistronic DNA vaccine against porcine rotavirus, which carried the rotavirus nonstructuralprotein 4 (NSP4) and VP7 genes simultaneously. Using a balanced lethal system, the kanamycinresistance gene of expressing eukaryotic plasmids pVAX1 and pVAXD were replaced by theaspartate β-semialdehyde dehydrogenase(asd) gene. The NSP4 cleavage product (259–525)of rotavirus OSU strain and VP7 full-length genes were amplified by reverse transcriptionpolymerase chain reaction and then inserted into the eukaryotic single-expression plasmid,pVAX1-asd, and the eukaryotic dual-expression plasmid, pVAXD-asd, respectively. Therecombinant plasmids pVAX1-asd-NSP4, pVAX1-asd-VP7 and pVAXD-asd-NSP4-VP7 weretransformed into the attenuatedS. typhimuriumχ4550 strain by electrotransformation. Anindirect immunofluorescence assay of the expressed COS-7 cell suggested that the recombinantS. typhimuriumχ4550 strain was constructed successfully. The recombinantS.typhimuriumχ4550 strain was orally administered to BALB/c mice. The group immunised with dual- expression plasmids produced a significantly higher level of serum Immunoglobulin G (IgG)and intestinal Immunoglobulin A (IgA) than the group immunised with single-expressionplasmids. These results indicated that eukaryotic bicistronic plasmid DNA vaccines could besuccessfully constructed to enhance humoural, mucosal and cellular immune response againstrotavirus infection.

【 授权许可】

Unknown