The invention discloses a triple minRNA for resisting
virus infection of aids and the construction method thereof. The sequences of the triple minRNA are respectively designed according to
gene sequences of ccr5, pol and vif and combined to be miRNA oligomeric and
single strand DNA, then double-strand
DNA is formed through annealing, then double-strand
DNA is respectively inserted into the
expression vector of pcDNA<TM> 6.2-GW / EmGFPmiR of miRNA to built the mi
RNA expression
plasmid, and the jamming effect of mi
RNA expression
plasmid to
target gene can be detected through a Qpcr method. A miRNA string with the best
gene interference effect to ccr5, vif and pol is inserted into a carrier of pcDNA 6.2 <TM>-GW / EmGFP, so as to obtain a series-wound interference vector of pcDNA6.2<TM>-GW / EmGFP-ccr5-pol-vif, and then the miR-ccr5-pol-vif in the series-wound interference vector is transferred to a lentiviral vector pLenti6.3 / V5-DEST so as to construct pLenti6.3 / V5-DEST miR-ccr5-pol-vif. The recombinant lentiviral vector can product three miRNA at the same time to cause interference effects to
gene ccr5, gene pol and gene vif, so as to prevent the gene ccr5, the gene pol and the gene vif from expressing corresponding
protein. Therefore, the effect of controlling Hiv infection and replication can be achieved.