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NR-36461  Human respiratory syncytial virus , Respiratory Syncytial Virus (RSV) A2 Large Polymerase (L) Helper Plasmid, pA2-Lopt
(Proteins)
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Description: Respiratory Syncytial Virus (RSV) A2 Large Polymerase (L) Helper Plasmid, pA2-Lopt
Organism: Human respiratory syncytial virus
Biosafety Level: 1
Availability Status: In Stock
Store at: -80°C or colder
Contributor: BEI Resources
Comments:
Quantity limit per order for this item is 1. This item can be ordered twice a year. Orders over this limit will be sent to NIAID for approval before shipment.

NR-36461 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.

The L helper plasmid was constructed from codon-optimized RSV A2 L sequences. The codon-optimized cDNA sequences were synthesized and cloned into the pcDNA™ 3.1(+) mammalian expression plasmid (Life Technologies™ Invitrogen™). The plasmid was produced in Escherichia coli, strain 10-beta (a DH10B derivative, New England BioLabs®) and extracted using a Endo-Free Plasmid Maxi Kit (Qiagen).

Each vial contains 0.5 µg of plasmid DNA in RNase/DNase-free 10 mM Tris-HCl, 1 mM EDTA buffer (pH 8).

Recombinant RSV was produced by co-transfection of BHK-21 clone BSR T7/5 cells with pSynkRSV-I19F, a BAC plasmid containing RSV A2-line19F antigenomic DNA and the gene for the far-red fluorescent protein monomeric Katushka 2 (mKate2) to enable detection of infection through fluorescence, (NR-36460) and four helper plasmids encoding sequence-optimized genes from RSV strain A2: large polymerase (L) (NR-36461), nucleoprotein (N) (NR-36462), phosphoprotein (P) (NR-36463) and matrix 2-1 protein (M2-1) (NR-36464). RSV rescue and infection could be detected by red fluorescent syncytia.
Citations: Acknowledgment for publications should read "The following reagent was obtained through BEI Resources, NIAID, NIH: Respiratory Syncytial Virus (RSV) A2 Large Polymerase (L) Helper Plasmid, pA2-Lopt, NR-36461."
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  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.

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