携带pri-mir-20a的慢病毒表达载体的构建

in this study, the pri-mir-20a amplified by PCR is inserted into the Bam H Ⅰ/Hind Ⅲ digested pcDNA3.0-H1 to generate recombinant pcDNA3.0-H1 pri-mir-20a. H1-pri-mir-20a is then cloned into lentiviral expression vector pdc-SEW to generate pdcH1 pri-mir-20a-SEW. Finally, pCMV8.91, pMD2. VSV. G, pdcH1-pri-mir-20a-SEW are co-transfected cells of 293FT and the expression of green fluorescent protein reaches 80% in 48h. The result shows that the successful construction of lentivirus vector pdcH1-pri-mir-20a-SEW provides the basis for further study of biological functions and the targets of miR-20a.