NR-19813_扎伊尔埃博拉病毒,载体pcDNA3.1含有融合至埃博拉病毒的?-内酰胺酶,扎伊尔VP40(Bla-VP40)(质粒/载体）

NR-19813??Zaire ebolavirus, Vector pcDNA3.1 Containing ?-Lactamase Fused to Ebolavirus, Zaire VP40 (Bla-VP40)(Plasmid/Vectors)|Zaire ebolavirus|Vector pcDNA3.1 Containing ?-Lactamase Fused to Ebolavirus, Zaire VP40 (Bla-VP40)|-20°C or colder|A Garcia-SastreAcknowledgment for publications should read "The following reagent was obtained through BEI Resources, NIAID, NIH: Vector pcDNA3.1 Containing β-Lactamase Fused to Ebolavirus, Zaire VP40 (Bla-VP40), NR-19813."|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. The viral matrix protein VP40 gene from ebolavirus (EBOV), Zaire was synthesized by multiple rounds of overlapping PCR based on the EBOV, Zaire genome sequence (GenBank accession L11365). The ?-lactamase gene was PCR amplified from the pcDNA3.1 vector (Invitrogen?) and fused to the N-terminal of VP40 by a short linker sequence (GSGGGSGGT) to create a modified ?-lactamase-VP40 fusion protein (Bla-VP40) which was subcloned into pcDNA3.1. The modified ?-lactamase lacks the N-terminal 24 amino acid secretion signal and His24 was substituted by Asp to create an optimal Kozak consensus sequence. The plasmid was produced in Escherichia coli 5-alpha F?I^q cells (New England Biolabs^?) and extracted using a QIAGEN^? plasmid DNA extraction kit.VP40 drives the budding of filovirus particles. 293T cells co-transfected with NR-19813 and the EBOV glycoprotein (NR-19814) or the Marburg virus (MARV) glycoprotein (NR-19815) produce EBOV or MARV virus-like particles (VLPs), respectively. Fusion of these VLPs with target cells can be detected by monitoring ?-lactamase activity using a fluorogenic substrate, permitting study of the cell entry steps of these highly pathogenic viruses without the need for BSL-4 containment.NR-19813 has been qualified for use in bacterial transformations.Each vial contains 20 to 50 ng of plasmid DNA in TE buffer (10 mM Tris-HCl, 1 mM EDTA, pH 7.0).