Cloning and Expression of TAT-apoptin and Its Activity Study in vitro

A basic peptide derived from human immunodeficiency virus HIV-1 TAT peptide(positions 47-57) can translocate through the cell membranes,the characteristics of which are utilized for the deli-(very) of exogenous proteins into cells. The coding sequence for the TAT-apoptin was amplified by polymerase chain reaction(PCR),then inserted into pET28b vector and highly expressed in Escherichia coli BL21(DE3).The protein expressed in inclusion body was purified by Ni-NTA under denaturing condition.(Apoptosis) activity of the TAT apoptin was evaluated by MTT.Results indicated that HeLa cells were highly sensitive to the purified TAT apoptin.