重组α-病毒的制备和鉴定

Preparation of recombinant alpha-virus

目的 探讨从DNA水平制备重组α 病毒的新途径 ,寻找更简便的制备重组α 病毒疫苗的新方法。方法 将表达β 半乳糖苷酶的载体和包装载体用磷酸钙法共转染BHK包装细胞 ,收集并纯化上清中的病毒。再用该病毒在体外感染BHK细胞 ,鉴定所制备病毒滴度和目的基因的表达情况。结果 用本方法制备出了较高效价的重组病毒 ,并能在哺乳细胞中很好地表达。结论 从DNA水平可以制备出较高滴度的重组α 病毒 。

Objective To explore a new simpler method for the preparation of recombinant alpha virus as a novel vaccine at the DNA level. Methods Plasmids expressing β gal protein and helper plasmids were transfected into BHK cells. Virus in culture supernatant of the transfected BHK cells were collected and purified and used to infect BHK cells in vitro to identify the expression of target gene and the titre of the recombinant virus. Results Recombinant virus with high titre, prepared by this method, could be expressed well in mammalian cells in vitro . Conclusion High titre recombinant alpha virus can be produced at the DNA level and this method can be applied for vaccine preparation and gene therapy.