Abstract:
Plaque assay method enables the quantification of infectious baculovirus when defined as plaque forming units (PFU). It allows to determine the amount of infectious virus needed to infect the cells at a specific multiplicity of infection (MOI). Serial dilutions of baculovirus stock are added to the Sf9 cells monolayer followed by addition of 5% Agarose overlay. Six days after infection clear infection halos are observed using a neutral red solution. Here we describe the quantification of recombinant baculovirus expression vector (rBEV) carrying a transgene in an rAAV expression cassette. Reproducible quantification of PFU is obtained with this method.
摘要:
当定义为噬斑形成单位(PFU)时,噬斑测定方法能够定量感染性杆状病毒。它允许确定以特定的感染复数(MOI)感染细胞所需的感染性病毒的量。将连续稀释的杆状病毒原液添加到Sf9细胞单层中,然后添加5%琼脂糖覆盖。感染后6天,使用中性红溶液观察到明显的感染光晕。在这里,我们描述了在rAAV表达盒中携带转基因的重组杆状病毒表达载体(rBEV)的定量。用这种方法获得PFU的可重复定量。