绵羊肺腺瘤病毒 SU 蛋白与 Hyal-2蛋白的结合域研究

摘要

试验证明绵羊的 Hyal-2不仅可以与绵羊肺腺瘤病毒的囊膜蛋白 Env 结合，也可以单独与绵羊肺腺瘤病毒 SU 蛋白亚基结合。为了进一步研究绵羊 Hyal-2与 SU 蛋白可能结合的区域，应用在线生物学软件预测了 SU 蛋白的膜外区（对应编码基因序列238 bp～867 bp）将之前已经构建的缺失型 SU 蛋白真核表达载体 pEGFP-C1-（su1-su5）与 pDsRed-monomer-N1-hyal-2共转染293T 细胞，运用激光共聚焦方法结合免疫共沉淀方法确定 SU 蛋白和 Hyal-2蛋白可能结合区域。激光共聚焦显微镜观察结果表明，SU1-SU5与 Hyal-2均存在细胞内共定位。免疫共沉淀方法检测缺失型蛋白 SU1-SU5与绵羊 Hyal-2均不存在相互作用。su 基因所缺失区域238 bp～867 bp 对于 SU 蛋白与受体的结合都是必不可少的。%The experimental results have shown that the Hyal-2 can not only be combined with the Env pro-tein of Ovine pulmonary adenomatosis virus(OPAV),but also can be combined with the SU protein of OPAV.To design the experiment for further studying the possible binding regions of Hyal-2 and SU pro-tein,the extracellular domain of SU protein was predicted by using online biology software(238 bp-867 bp).pEGFP-C1-(su1-su5)and pDsRed-monomer-N1-hyal-2 vectors were cotransfected into 293T cells. The possible binding area between SU protein and Hyal-2 protein by using laser confocus and co-immuno-precipitation methods was determined.The results showed that SU1-SU5 and Hyal-2 were co-located in the cells,there was no interaction between the SU1-SU5 and the Hyal-2 of sheep by using the method of co-im-munoprecipitation.The deleted region 238 bp-867 bp of the su gene is essential for the binding of SU with the receptor.