NR-36460_人类呼吸道合胞病毒

NR-36460??Human respiratory syncytial virus , Bacterial Artificial Chromosome Plasmid pSynkRSV-I19F Containing Antigenomic cDNA from Respiratory Syncytial Virus (RSV) A2-Line19F(Plasmid/Vectors)|Human respiratory syncytial virus|Bacterial Artificial Chromosome Plasmid pSynkRSV-I19F Containing Antigenomic cDNA from Respiratory Syncytial Virus (RSV) A2-Line19F|-80°C or colder|BEI ResourcesAcknowledgment for publications should read "The following reagent was obtained through BEI Resources, NIAID, NIH: Bacterial Artificial Chromosome Plasmid pSynkRSV-I19F Containing Antigenomic cDNA from Respiratory Syncytial Virus (RSV) A2-Line19F, NR-36460."|Quantity limit per order for this item is 2. This item can be ordered twice a year. Orders over this limit will be sent to NIAID for approval before shipment. NR-36460 is a component of a bacterial artificial chromosome (BAC)-based RSV rescue system that allows RSV infection to be monitored by fluorescence and is an important tool in RSV vaccine research and mutagenesis studies. Antigenomic cDNA of RSV A2-line19F was synthesized in three segments which were cloned sequentially into pKBS2 (BAC vector) to generate the pSynkRSV-I19F plasmid. The gene for the far-red fluorescent protein monomeric Katushka 2 (mKate2) was included in the RSV antigenomic DNA to enable detection of infection through fluorescence. The plasmid was produced in Escherichia coli, strain SW102 cells and extracted using a NucleoBond^? BAC 100 (Macherey-Nagel) plasmid DNA purification kit. Each vial contains 0.5 ?g of plasmid DNA in RNase/DNase-free 10 mM Tris-HCl buffer (pH 8.5). The vial should be centrifuged prior to opening. Note: The protocol provided (refer to Product Information Sheet) requires 0.8 ?g of pSynkRSV-I19F; thus 2 vials of NR-36460 are required per transfection. Recombinant RSV was produced by co-transfection of BHK-21 clone BSR T7/5 cells with NR-36460 and four helper plasmids containing sequence-optimized genes for RSV strain A2 large polymerase (NR-36461), nucleoprotein (NR-36462), phosphoprotein (NR-36463) and matrix 2-1 protein (NR-36464). RSV rescue and infection was detected by red fluorescent syncytia.