Broekhuijsen M P, Blom T, Kottenhagen M, Pouwels P H, Meloen R H, Barteling S J, Enger-Valk B E
Vaccine. 1986 Jun;4(2):119-24. doi: 10.1016/0264-410x(86)90050-2.
Part of the genome of foot-and-mouth disease virus (FMDV) type 01,BFS, including the sequence encoding the capsid polypeptide VP1, was cloned in Escherichia coli following a new cloning strategy. The clone containing the VP1 sequence was used for the construction of two expression plasmids encoding VP1 fusion proteins. Subsequently, substantial amounts of the two VP1-beta-galactosidase fusion proteins, containing either one (amino acid region 140-160) or two (amino acid regions 140-160 and 200-213) antigenic determinants of the virus, were synthesized by E. coli bacteria. The protein containing the amino acid region 140-160 of VP1 fused to beta-galactosidase efficiently induced antibodies in rabbits specifically reacting with FMDV type 01,BFS. The same protein was also capable of eliciting neutralizing antibodies. The fusion protein containing both antigenic determinants did not efficiently induce antibodies reacting with FMDV.
采用一种新的克隆策略,将01型口蹄疫病毒(FMDV)BFS株的部分基因组,包括编码衣壳多肽VP1的序列,克隆到了大肠杆菌中。含有VP1序列的克隆用于构建两个编码VP1融合蛋白的表达质粒。随后,大肠杆菌合成了大量的两种VP1-β-半乳糖苷酶融合蛋白,其中一种含有该病毒的一个抗原决定簇(氨基酸区域140 - 160),另一种含有两个抗原决定簇(氨基酸区域140 - 160和200 - 213)。与β-半乳糖苷酶融合的含有VP1氨基酸区域140 - 160的蛋白能有效诱导兔体内产生与01型口蹄疫病毒BFS株特异性反应的抗体。同样的蛋白也能够引发中和抗体。含有两个抗原决定簇的融合蛋白不能有效诱导与口蹄疫病毒发生反应的抗体。