[Studies of the resistance of the recombinant alphavirus RNAs containing dengue-2 PrM gene to virus infection].
The amplified PrM gene of dengue-2 virus was cloned into the downstream SP6 promoter-pSFV vector and the recombinant plasmid (pSF.rM2) DNA which contained sense- or antisense-PrM gene, was selected. pSF.rM2 DNA and helper DNA linearized by the enzyme SpeI digestion were both transcribed in vitro into recombinant RNAs which contained the capping analog on the 5'-end and contaansfected into BHK cells by electroportation. The the transfected host cells were challenged with dengue-2 virus and the resistant efficiency of recombinant virus RNAs containing sense- or antisense-PrM gene to virus infection were observed, respectively. The recombinant plasmids (pSFV-PrM) containing sense- or antisense-PrM gene were selected with determination of the nucleotide sequence. The recombinant virus particles were obtained with recombinant RNA and helper RNA co-transfected into BHK cells. Host cells transfected with antisense-PrM RNA derived complete resistance to dengue-2 virus replication and the efficiency was higher than that of the recombinant virus RNA containing sense-PrM gene.
Yu
M
M
Department of Virology, Institute of Microbiology and Epidemiology, Beijing 100850, China.
Qin
E
E
Zhao
W
W
Hu
Z
Z
Yuan
X
X
D000524:Alphavirus / Q000235:genetics*; D000818:Animals; D002478:Cells, Cultured; D006224:Cricetinae; D003716:Dengue Virus / Q000235:genetics* / Q000502:physiology; D005347:Fibroblasts / Q000166:cytology / Q000821:virology; D005814:Genes, Viral; D005822:Genetic Vectors; D007668:Kidney / Q000166:cytology / Q000821:virology; D008565:Membrane Proteins / Q000235:genetics* / Q000494:pharmacology; D010957:Plasmids; D012313:RNA / Q000235:genetics / Q000494:pharmacology; D012367:RNA, Viral / Q000235:genetics / Q000494:pharmacology; D014779:Virus Replication / Q000187:drug effects*
D008565:Membrane Proteins; D012367:RNA, Viral; C039919:RNA, recombinant; D012313:RNA