Klepfer S, Reed A P, Martinez M, Bhogal B, Jones E, Miller T J
Department of Molecular Biology, Smithkline Beecham Animal Health, King of Prussia, Pennsylvania, USA.
Adv Exp Med Biol. 1995;380:235-41. doi: 10.1007/978-1-4615-1899-0_38.
The spike gene of the feline enteric coronavirus (FECV), strain FECV-1683, was PCR amplified from total RNA extracted from FECV-infected cells and its sequence determined. A primary translation product of 1454 amino acids is predicted from the nucleotide sequence, containing a N-terminal signal sequence, a C-terminal transmembrane region and 33 potential N-glycosylation sites. The sequence shares 92% homology with the previously published feline infectious peritonitis virus, strain WSU-1146; however, several regions were identified that distinguished FECV from Feline Infectious Peritonitis virus, FIPV. The full length FECV S gene was cloned and expressed in vaccinia virus. Recombinants produced a 200 kD protein which was recognized by sera from cats infected with FIPV. When kittens were immunized with the vaccinia/FECV S recombinant, neutralizing antibodies to FIPV were induced. After challenge with a lethal dose of FIPV, the recombinant vaccinated animals died earlier than control animals immunized with vaccinia virus alone.
从感染猫肠道冠状病毒(FECV)-1683毒株的细胞中提取总RNA,通过PCR扩增该毒株的刺突基因并测定其序列。根据核苷酸序列预测,其初级翻译产物为1454个氨基酸,包含一个N端信号序列、一个C端跨膜区域和33个潜在的N-糖基化位点。该序列与先前发表的猫传染性腹膜炎病毒WSU-1146毒株具有92%的同源性;然而,也发现了几个将FECV与猫传染性腹膜炎病毒(FIPV)区分开来的区域。克隆了全长FECV S基因并在痘苗病毒中表达。重组体产生了一种200 kD的蛋白质,该蛋白质可被感染FIPV的猫的血清识别。用痘苗/FECV S重组体免疫小猫后,可诱导产生针对FIPV的中和抗体。在用致死剂量的FIPV攻击后,接种重组体的动物比仅接种痘苗病毒的对照动物死亡更早。