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HRP-20053 is a full-length molecular clone of infectious and replication-competent simian-human immunodeficiency provirus. This clone contains an amino acid residue at Env position 375 that supports virus entry and replication in primary rhesus CD4 T cells. SHIV.1086.375W.dCT is an isogenic mutant of SHIV.1086.375S.dCT generated by changing 1086 Env375 residue (GenBank: MW410739) from Ser to Trp. SHIV.1086.375W.dCT showed increased infectivity and replication kinetics in vitro in Indian rhesus macaque CD4+ T cells and in vivo in Indian rhesus macaques. The plasmid encodes full-length, replication-competent SHIV in a pBR322 backbone. The beta-lactamase gene, bla, provides transformant selection through ampicillin resistance in Escherichia coli (E. coli). The resulting size of the plasmid is 14,890 base pairs. The plasmid sequence and map are provided on the NIH HIV Reagent Program webpage.
Note: Bacterial transformation and large-scale preparation of plasmid DNA are tricky due to plasmid instability. The contributors have developed a protocol to prevent this by growing bacteria for shorter time periods and at lower temperatures.Each vial contains plasmid DNA in TE buffer (10 mM Tris-HCl, 1 mM EDTA). The vial should be centrifuged prior to opening. Note: The contents of the vial should be used to transform the plasmid in E. coli prior to mammalian expression.
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For more information on the necessary compliance requirements associated with the biological materials provided by NIH HIV Reagent Program, please select this link: Compliance Requirements.