猪口蹄疫病毒多抗原表位重组腺病毒的构建与鉴定

本研究设计构建了含有猪O型口蹄疫病毒VP1（21—60）-（141-160）-（200—213）位氨基酸的基因的重组腺病毒质粒pAd-VP，经PacI酶切后转染HEK-293A细胞，3次噬斑纯化获得了重组腺病毒rAd—VP。该重组腺病毒于HEK-293A细胞连续传代至20代效价稳定，TCID50为10^-10／mL。RT—PCR检测证明目的基因在mRNA水平上可有效表达；应用O型口蹄疫病毒标准阳性血清进行间接荧光抗体试验，在rAdVP感染的HEK-293A细胞的胞质可见清晰荧光。证明该重组腺病毒对VP1（21-60）-（141—160）-（200—213）位氨基酸的基因进行了成功的表达，从而为FMDV多抗原表位腺病毒活载体疫苗的研究奠定了基础。

Construction and Indentification of Recombinant Adenovirus Containing Multiple Antigen Epitopes of Swine Foot-and-Mouth Disease Virus

A recombinant replication-defective Human adenovius serotype 5 plasmid pAd-VP,containing amino acids(21-60)-(141-160)-(200-213) coding region of swine foot-and-mouth disease virus serotype O VP1,was constructed using the method of homologous recombination in E.coli BJ5183.After the recombinant plasmid pAd-VP linearized with PacI transferred into HEK-293A cell,the recombinant virus was isolated and purified in HEK-293A cells by three times plaque purification.This recombinant adenovirus could be stably passaged in HEK-293A cells and TCID_50 was 10~-10/mL.The mRNA of the gene was detected by RT-PCR.Obvious fluorescence was observed in the HEK-293A cell stained with standard serum of FMDV serotype O in IFA.The recombinant adenovirus could express the antigen of FMDV.The adenovirus containing multiple antigen epitopes of FMDV can be furtherly used to develop the FMDV vaccine.