Construction of recombinant baculovirus expressing TGEV spike protein S2 and the preliminary researc

• 作 者: Li Wang Xianjin Dai Han Song Yue Zhou Cuifang Ye Yang Hu Zhenhui Song
• 机 构: Department of Veterinary Medicine,Southwest University,Rongchang Campus,Chongqing 402460,P.R.China
• 出 处: 第七届中国畜牧科技论坛
• 发表日期: 2016年4期
• 关键词: Transmissible gastroenteritis virus S gene baculovirus expression system immune

　　S protein is the only structure protein to induce TGEV-neutralizing antibodies.At the N terminus of the S protein,there are four antigenic sites,A,B,C,and D,which have been shown to be involved in the stimulation of neutralizing antibodies.In this study we constructed recombinant baculoviruses expressing functional antigenic sites D and A sites of spike gene of transmissible gastroenteritis coronavirus(TGEV)with size of 1034bp,was amplified by using PCR assays and cloned into pFastBacTMDual vector,after transposition and transfection,the recombinant baculoviruses rBacmid-S2 was obtained.Then the recombinant baculoviruses rBacmid-S2 was identified with molecular masses of 37.9KDa by SDS-PAGE.Mice were immunized intramuscularly,orally with recombinant baculovirus rBacmid-S2 cell lysis buffer 7×107TCID50/100μL per mice.Sera and feces samples were collected separately on days 0,14,28,42 to detect levels of IgG and sIgA against TGEV by ELISA assays.The results indicated that the recombinant baculovirus rBacmid-S2 could significantly induce humoral(specific IgG)and mucosal(sIgA)in mice and the antibody levels of intramuscular route higher than the orally.The results preliminary confirmed that the recombinant baculovirus expressing S2 gene of TGEV could induce a certain mucosal and humoral immune responses in mice.