반응 #663664

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penicillin

streptomycin

CO2

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Deoxynojirimycin

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후처리

1. 1
   기타BVDV-free MDBK cells (CCL 22) were obtained from the American Type Culture Collection
2. 2
   온도heat inactivated horse serum (Invitrogen)
3. 3
   workup.ADDITIONFor infections, virus inoculum was added in complete medium
4. 4
   기타the inoculum was then removed
5. 5
   세척the cells washed once with medium and fresh medium
6. 6
   workup.ADDITIONcontaining compounds
7. 7
   workup.ADDITIONadded
8. 8
   기타Virus stocks were prepared by freeze-thawing the infected cells and culture supernatant three times
9. 9
   workup.WAITfollowed by centrifugation at 1,000 g for 5 min
10. 10
    기타stored at −80° C
11. 11
    기타WNV was obtained from a cDNA clone of a human 2002
12. 12
    기타isolate from Texas

실험 절차

Cells and viruses. BVDV-free MDBK cells (CCL 22) were obtained from the American Type Culture Collection and propagated in DMEM/F12 essential medium supplemented with penicillin (500 U/ml), streptomycin (500 U/ml), and 10% heat inactivated horse serum (Invitrogen). Cells were maintained in a humidified incubator at 37° C. with 5% CO2. BVDV (NADL strain). For infections, virus inoculum was added in complete medium and adsorbed for 1 hour at 37° C., the inoculum was then removed, the cells washed once with medium and fresh medium containing compounds added. Virus stocks were prepared by freeze-thawing the infected cells and culture supernatant three times followed by centrifugation at 1,000 g for 5 min. Stock titers were determined, and stocks were aliquoted and stored at −80° C. WNV was obtained from a cDNA clone of a human 2002 isolate from Texas; virus obtained from BHK cells electroporated with the in vitro synthesized RNA from this cDNA clone was passaged in Vero cells before use in antiviral assays. Dengue virus (DV) serotype 2 was a New Guinea C virus that had been passaged 28 times in suckling mouse brain, twice in Vero cells, and once in C6/36 mosquito cells.

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