By Peggy Geluykens, Koen Van Acker, Johan Vingerhoets, Christel Van den Eynde (auth.), Edwin Yunhao Gong (eds.)
Although antiviral medications were effectively constructed for a few viral illnesses, there continues to be a transparent, unmet scientific have to improve novel antiviral brokers for the keep an eye on and administration of many viruses that at present don't have any or restricted therapies in addition to a necessity to beat the constraints linked to the prevailing antiviral medications, akin to antagonistic results and emergence of drug-resistant mutations. the second one version of Antiviral equipment and Protocols features:
- All chapters are new and written through specialists within the box, reflecting the key fresh technical advances in antiviral study and discovery.
- This version specializes in many very important human viruses, resembling human immunodeficiency virus style 1 (HIV-1), hepatitis viruses (hepatitis B and C viruses), herpes viruses, human respiration syncytial virus (RSV), and influenza virus, whereas additionally that includes a few vital rising viruses, reminiscent of dengue virus, West Nile virus, and chikungunya virus.
- As a quantity within the hugely profitable Methods in Molecular Biology sequence, chapters contain introductions to their respective issues, lists of the required fabrics and reagents, step by step, without difficulty reproducible laboratory protocols, and tips about troubleshooting and heading off identified pitfalls.
Comprehensive and state of the art, Antiviral tools and Protocols, moment Edition will function a superb laboratory reference for pharmaceutical and educational biologists, medicinal chemists, and pharmacologists in addition to for virologists within the box of antiviral study and drug discovery.
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Extra info for Antiviral Methods and Protocols
11. 12. 13. 14. 15. 16. 17. of a viral DNA end by HIV-1 integrase. Biochemistry 38:8458–8468 Deprez E, Tauc P, Leh H et al (2000) Oligomeric states of the HIV-1 integrase as measured by time-resolved fluorescence anisotropy. Biochemistry 39:9285–9294 He HQ, Ma XH, Liu B et al (2007) Highthroughput real-time assay based on molecular beacons for HIV-1 integrase 3′-processing reaction. Acta Pharmacol Sin 28:811–817 Grobler JA, Stillmock KA, Hazuda D (2009) Scintillation proximity assays for mechanistic and pharmacological analyses of HIV-1 integration.
The nucleotide sequence obtained from the patient sample is aligned against the wild-type, reference HXB2 HIV-1 sequence (Genbank Accession number K03455) to construct a nucleotide contig. The PR–RT contig contains the complete 99 amino acids of the PR gene and the first 400 amino acids of the RT gene. The IN contig covers the complete 289 amino acids of the IN gene and is generated by assembling the RT–IN sequence that corresponds to the last 153 amino acids of RT gene, containing the 120 amino acids of the RNase H domain, and the complete IN gene.
4. Incubate the plates for 4 h at 37 °C (see Note 2). 5. Add 100 µL of stop solution (provided in the kit) to each well of row A. 6. Seal the plates with Topseal-A 384 (see Note 3). 7. Measure the plates using the TopCount: 2 min delay time, exposure time = 1 min/well. 8. Determine the optimal RT concentration based on a concentration that gives 2,000–4,000 counts per minute (CPM) (see Note 4). 2 Scintillation Proximity Assay 1. Prepare fourfold serial dilutions of test compounds in RPMI/10 % FBS at 2× final concentrations and transfer 25 µL of each test concentration to a 96-well MaxiSorp™ plate (2 % DMSO) (see Notes 5 and 6).
Antiviral Methods and Protocols by Peggy Geluykens, Koen Van Acker, Johan Vingerhoets, Christel Van den Eynde (auth.), Edwin Yunhao Gong (eds.)