慢病毒相关质粒

pNL4-3.Luc.R-E-质粒 HIV骨架假病毒报告载体

发布时间:2021-01-15 20:31:06 | 浏览次数：

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Firefly luciferase gene was inserted into the pNL4-3 nef gene. Two frameshifts (5鈥?Env and Vpr aa 26) render this clone Env- and Vpr-.

Competent for a single round of replication. Requires cotransfection with env expression vector to produce infectious virus. Virus can be produced by transfecting 2 x 106 293 or 293T cells with 20 ug of the NL4-3 DNA, or with 10 ug NL4-3 DNA and 10 ug env expression vector DNA. Transfections can be performed in a 10 cm2 tissue culture dish using standard calcium phosphate protocols. Virus is typically harvested 48 hours post-transfection. Infections should be performed in a total volume of 0.5 ml. Amphotropic pseudotypes generally have much higher infectivity than those bearing HIV-1 env. Cultures infected with the luciferase viruses can be lysed 2-5 days post-infection and assayed using commercial lysis buffer and luciferase reagents