Figure 2.

Construction and transfection of dual EGFP/shRNA expression vectors. (A) Buffalo, bovine and human 7SK/U6 promoter fragments were amplified with restriction enzymes sites XbaI/SacI introduced by the primers, and ligated into the 5′ MCS (multi-cloning site) of the pMCS-shEGFP and pMCS-sh1864 vectors. The core fragments in the vectors, featuring the MCS and sense, loop, antisense and pol III terminator (TTTTTT), were chemically synthesized and ligated with framework plasmid pMD18-T; (B) Fluorescence microscopy images of buffalo fetal fibroblasts (BFF) cells transfected with the reporter vector (pEGFP-N1) only, or co-transfected with the reporter and various shEGFP expression plasmids as indicated in each image.