کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
6141004 | 1227192 | 2013 | 12 صفحه PDF | دانلود رایگان |
- Heparan sulfate- and receptor-binding motifs of DENV2 envelope protein were mutated.
- Four mutant viruses were isolated-all could fuse C6/36 cells.
- Two of these mutants were lethal in Vero cells without further modification.
- Lethal mutations were KK291/295EV and KKK305/307/310EEE.
- Cell attachment was implicated as the replication block for both mutants.
Using an infectious cDNA clone we engineered seven mutations in the putative heparan sulfate- and receptor-binding motifs of the envelope protein of dengue virus serotype 2, strain 16681. Four mutant viruses, KK122/123EE, E202K, G304K, and KKK305/307/310EEE, were recovered following transfection of C6/36 cells. A fifth mutant, KK291/295EE, was recovered from C6/36 cells with a compensatory E295V mutation. All mutants grew in and mediated fusion of virus-infected C6/36 cells, but three of the mutants, KK122/123EE, E202K, G304K, did not grow in Vero cells without further modification. Two Vero cell lethal mutants, KK291/295EV and KKK307/307/310EEE, failed to replicate in DC-SIGN-transformed Raji cells and did not react with monoclonal antibodies known to block DENV attachment to Vero cells. Additionally, both mutants were unable to initiate negative-strand vRNA synthesis in Vero cells by 72Â h post-infection, suggesting that the replication block occurred prior to virus-mediated membrane fusion.
Journal: Virology - Volume 441, Issue 2, 5 July 2013, Pages 114-125