胎牛血清不同组分对昆虫杆状病毒复制的影响

The affect of different components of FBS on BAC virus replication

关键词:  FBS  血清  昆虫杆状病毒  SF9细胞

Key words:  FBS  Serum  BAC  SF9 cell

摘要:

为探讨胎牛血清（FBS）中的不同组分对昆虫杆状病毒（BAC）复制的影响，找到血清中能提高病毒复制效果的有效组分，对亲和凝胶层析柱法分离获得血清中的四个混合组分（S1、S2、S3和S4）进行了病毒培养实验。在6孔板上接种SF9细胞，培养24 h，接种0.01MOI的BAC-GFP病毒，按照0、0.3%、0.6%、1.3%、2.5%、5%的浓度梯度加入6孔板中进行病毒培养。分别在24 h、48 h、72 h、96 h进行荧光拍照、观察。96 h收集病毒上清，进行空斑形成实验测定病毒滴度。用收集的病毒感染SF9细胞，72 h流式检测病毒的复制情况。结果显示：血清样本S1在浓度1.3%以上时，对于BAC的复制有促进作用；S2没有促进效果，反而随着S2的浓度上升而出现抑制的情况；S3在浓度2.5%以上时，同样有着明显促进作用；S4的促进作用不显著。试验表明，与原胎牛血清相比，添加单一血清组分S1对提高BAC病毒的复制效果更显著。

Abstract:

To study the effect of different components of FBS on BAC virus replication，four components of FBS were obtained by affinity chromatography. SF9 cells were seeded into six well cell culture plates. SF9 cells were infected by 0.01MOI BAC virus in different concentrations (0、0.3%、0.6%、1.3%、2.5%、5% of FBS) at 24 hours. And The photos of cells were taken by Fluorescence microscope at 24、48、72、96 hours, respectively. Virus titer was detected at 96 hours. SF9 cells were infected by BAC virus, and virus replication was determined at 72 h. Results：S1 can significantly promote the BAC virus replication at concentration which is higher than 1.3%；S2 has no effect on BAC virus replication, but can inhibit its replication as the concentration goes down；S3 can also promote the BAC virus replication at a concentration which is higher than 2.5%；S4 has no influence on the BAC virus replication. In conclusion：In contrast to the original serum，S1 can significantly contribute to BAC virus replication.