登革2型PrM基因的重组病毒对不同型别登革病毒复制的阻断作用

The Blocking of Dengue Virus Replication by the Recombinant Alphavirus Containing Dengue-2 PrM Gene

观察登革 2型PrM基因的pSFV重组甲病毒抗该型病毒的作用 ,进一步探讨登革 2型PrM基因的这种重组病毒对其它 3个血清型登革病毒复制的阻断作用 .采用体外转录和电穿孔 ,分别将构建的含正、反义PrM基因的重组质粒DNA和辅助载体DNA转录成RNA ,然后将这两种RNA共转染BHK细胞 ,进而包装成重组病毒颗粒 .再将激活的重组病毒感染细胞 ,分别用不同型病毒进行攻击 .然后通过免疫荧光法 ,观察对登革病毒复制的阻断作用 .结果表明 ,含登革 2型PrM基因的重组病毒不仅可阻断登革 2型病毒的复制 ,同样具有抑制其他 3个型病毒复制的能力 ,且抗登革 1、4型病毒的复制作用强于抗登革 3型病毒的作用 .用 10 3 TCID50 剂量的登革病毒攻击 ,含反义PrM基因的重组病毒可完全阻断登革 1、3、4型病毒的复制 .但含正义PrM基因的重组病毒对登革 3型病毒的复制不能完全阻断 .

Based on the resistance of the recombinant alphavirus containing dengue-2 PrM gene against dengue virus, replications of three other dengue serotype viruses were blocked by this recombinant virus. The recombinant plasmid (pSF-rM 2) DNA containing sense- or antisense-PrM gene and helper vector DNA were transcribed into RNAs in vitro, respectively. These transcribed RNAs were cotransfected into the culture of BHK cells by electroporation and were packaged into the recombinant virus particles. After these recombinant viruses were activated, BHK cells were infected by them and challenged with other three type dengue viruses. The blocking of dengue virus replication was observed by immunofluorescence. The results show that the recombinant deugue-2 PrM alphavirus can also inhibit replications of types 1,3 and 4, and the resistance of replications of type 1 and 4 were higher than that of type 3 virus. The recombinant antisense-PrM alphavirus can completely block replications of dengue type 1,3 and 4 viruses. But the recombinant sense-PrM alphavirus can not completely block replication of type 3 virus, when the transfected host cells were challenged with dengue viruses at 10 3 TCID 50 . These results might lay a foundation for exploring a new way to cure the dengue disease.