Construction of recombinant vector containing fusion gene NT ₄-ant-shepherdin[79-87]

Objective: To investigate survivin as an anticancer therapeutic target by use of shepherdin [79-87], a novel peptide carrying the survivin sequence from Lys-79 through Leu-87, we constructed an recombinant vector containing fusion gene NT₄-Ant-Shepherdin [79-87]. Methods: The gene of Ant-Shepherdin [79-87] was obtained by PCR and T-vector method. After cloned and digested with restricted enzyme, Ant-shepherdin [79-87] was inserted in PBV220/NT4 vector. The recombinant vector was transformed into the competent cell, E. coli DH5 α. The fusion gene of NT₄-Ant-Shepherdin [79-87] was identified by agarose gel electrophoresis (AGE). Results: DNA sequencing results verified that the sequence of Ant-Shepherdin [79-87] was consistent with what we had designed. After transformed E. coli DH5 α, a fragment of 321 bp was confirmed. Conclusion: The recombinant vector containing fusion gene NT ₄-Ant-Shepherdin [79-87] was successfully constructed in this experiment by molecular biology techniques, which provides the basis of further research of survivin for cancer gene therapy.