Crabb B S, Studdert M J
Centre for Equine Virology, School of Veterinary Science, University of Melbourne, Parkville, Vic, Australia.
Vet Microbiol. 1995 Sep;46(1-3):181-91. doi: 10.1016/0378-1135(95)00082-l.
A series of truncated equine herpesvirus 1 (EHV1) glycoprotein C (gC) molecules was examined for use as serodiagnostic antigens for EHV1 and EHV4. Small regions of EHV1 glycoprotein C, an immunodominant EHV1 glycoprotein, were expressed in Escherichia coli as glutathione S-transferase (GST) fusion proteins using the bacterial expression vector pGEX-2T. Sera obtained from horses, including sera from specific-pathogen-free (SPF) foals, following exposure to either EHV1, EHV4 or both viruses were used. Several of the fusion proteins were shown to encompass EHV1 specific epitopes while others encompassed strong, cross-reactive epitopes. One clone, termed pEC-3, produced a soluble and stable fusion protein which encompassed amino acids 107-275 of EHV1 gC. Strong cross-reactive epitopes on pEC-3 were localised to a region encompassed by amino acids 137 to approximately 152 while EHV1 specific epitope(s) were identified downstream of this region, i.e., approximately amino acids 152 to 275. E. coli expressed EHV1 gC polypeptides showed clear potential for use as diagnostic reagents for the detection of cross-reactive and type-specific EHV1 and EHV4 antibodies present in convalescent equine sera.
对一系列截短的马疱疹病毒1型（EHV1）糖蛋白C（gC）分子进行了检测，以用作EHV1和EHV4的血清学诊断抗原。EHV1糖蛋白C是一种免疫显性EHV1糖蛋白，其小区域利用细菌表达载体pGEX - 2T在大肠杆菌中作为谷胱甘肽S - 转移酶（GST）融合蛋白表达。使用从马获得的血清，包括来自无特定病原体（SPF）马驹的血清，这些马驹在接触EHV1、EHV4或两种病毒后采集血清。一些融合蛋白显示包含EHV1特异性表位，而其他融合蛋白包含强的交叉反应性表位。一个名为pEC - 3的克隆产生了一种可溶性且稳定的融合蛋白，其包含EHV1 gC的第107 - 275位氨基酸。pEC - 3上的强交叉反应性表位定位于第137位至约152位氨基酸所包含的区域，而EHV1特异性表位在该区域下游被鉴定出来，即大约第152位至275位氨基酸。大肠杆菌表达的EHV1 gC多肽显示出明显的潜力，可作为诊断试剂用于检测恢复期马血清中存在的交叉反应性和型特异性EHV1和EHV4抗体。
