Reverse genetics approach to study the molecular biology of rotavirus

我们构建并鉴定了一种表达VP4的重组轮状病毒，该病毒具有可被呋喃和胰酶识别的切割位点。出乎意料的是，VP4裂解位点突变病毒在没有外源蛋白酶的情况下不能进行多周期复制。此外，即使在胰酶存在的情况下，突变病毒的感染性也低于携带VP4的亲本病毒。这些结果表明，VP4的胞内被Furin裂解可能不利于轮状病毒的感染性。

We generated and characterized a recombinant rotavirus expressing cDNA-derived VP4 with a modified cleavage site recognized by furin as well as trypsin. Unexpectedly, the VP4 cleavage site mutant virus could not undergo multicycle replication without an exogenous protease. Furthermore, the mutant virus showed lower infectivity even in the presence of trypsin compared with the parental virus carrying authentic VP4.These results suggest that intracellular cleavage of VP4 by furin may be disadvantageous for rotavirus infectivity.