Objective To detect the function of I BHV-1 TKVgE-/ EGFP+ gene-deleted mutant vims strain, find a new method to prevent infectious bovine rhinotracheitis (IBR) , the charaction of TK-/gE-/ EGFP+ gene-deleted mutant virus strain was further studied. Methods Southern hybridization, western dot blot and Plaques test were employed in this investment. Results TK gene is deleted and the mutant cannot produce glycoprotein E when gE gene is deleted. The diameter of mutant strain much smaller than that of field strain, the mutant strain grows on MDBK cells and the virus titer can keep 106. OTCID50/0. 1 mL for more than 40 hours, which indicated that the mutant strain inherit instantly. Conclusion The gene-deleted mutant virus strain was successful constructed in this study, and the mutantrnvirus strain was stable. This will be useful for the further study of IBR.%目的 构建Ⅰ型牛痘疹病毒(BHV-1)的TK -/gE -双缺失突变株,检测其生物学功能,为治疗牛传染性鼻气管炎提供参考.方法 构建了带有EGFP表达盒的BHV-1 TK-/gE-双缺失突变株.通过southern杂交、蛋白质斑点印迹、空斑试验、MDBK细胞增殖实验等技术方法,对重组基因所构成病毒突变株的生物学特性进行鉴定.结果 Southern杂交及蛋白斑点印迹表明,课题组成功构建了带有EGFP表达盒的双缺失突变株；该突变株具有与野生株相当的繁殖力,但TK-/gE-成功缺失,毒力大幅减弱；BHV-1 TK-/gE-遗传稳定,不会返强.结论 本项目组成功构建了Ⅰ型牛疱疹病毒(BHV -1)的TK -/gE -双缺失突变株,该缺失株具有良好的安全性和稳定性,为该突变株进一步作为生物安全疫苗和多价病毒载体提供了依据,为预防和治疗牛传染性鼻气管炎提供了技术支持.
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