The prM-E-NS1 regions (820 bp-3438 bp in GZ01 KU820898; 927 bp-3544 bp in FSS13025 KU955593) were amplified from cDNA of infected cells. The prM-E-NS1 genes from the P6-740 strain (820 bp-3438 bp, HQ234499) and the Micronesian 2007 strain (820 bp-3438 bp, EU545988) were commercially synthesized (Genescript, China). All the genes were subcloned into a pcDNA3.1/V5-His/TOPO DNA vector (K4800-01, Invitrogen), and subsequently transfected by the X-treme Gene HP DNA transfection reagent (06366236001, Roche) for ectopically expression in 293T cells. The amount of NS1 was determined in the supernatants of the 293T cells at 48 hrs post-transfection. The cloning primers are shown in Extended Data Table 1 .
Pcdna3.1 v5 his topo dna vector
Manufactured by Thermo Fisher Scientific
The PcDNA3.1/V5-His/TOPO DNA vector is a plasmid-based expression vector used for the efficient cloning and expression of recombinant proteins in mammalian cell lines. It provides a simple and efficient method for the directional cloning of blunt-end PCR products, offering a streamlined approach to protein expression studies.
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Lab products found in correlation
2 protocols using pcdna3.1 v5 his topo dna vector
1
Amplification and Expression of DENV prM-E-NS1 Regions
2
Amplification and Expression of DENV prM-E-NS1 Regions
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The prM-E-NS1 regions (820 bp-3438 bp in GZ01 KU820898; 927 bp-3544 bp in FSS13025 KU955593) were amplified from cDNA of infected cells. The prM-E-NS1 genes from the P6-740 strain (820 bp-3438 bp, HQ234499) and the Micronesian 2007 strain (820 bp-3438 bp, EU545988) were commercially synthesized (Genescript, China). All the genes were subcloned into a pcDNA3.1/V5-His/TOPO DNA vector (K4800-01, Invitrogen), and subsequently transfected by the X-treme Gene HP DNA transfection reagent (06366236001, Roche) for ectopically expression in 293T cells. The amount of NS1 was determined in the supernatants of the 293T cells at 48 hrs post-transfection. The cloning primers are shown in Extended Data Table 1 .
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