Construction of the overexpressed lentiviral vector for targeting human TNFAIP1 gene / 中国医师杂志

ABSTRACT

Objective To construct a lentivirus vector over-expressing Tumor necrosis factor alpha induced protein 1 (TNFAIP1) gene and detect its expression level in vitro.Methods The full length of TNFAIP1 gene fragments were amplified by polymerase chain reaction (PCR).The pEZ-Lv105 vectors were digested by restriction endonuclease which was then linked to the full length of TNFAIP1 gene fragments by using T4DNA ligase.The plasmids were transfected into E.coli stbl3 and then we obtained the highly expressing positive clones by screening and identifying.The lentivirus vectors containing TNFAIP1 gene were transfected into 293T cells for package according to the packing kit manual.Results TNFAIP1 gene was amplified and successfully bound to the pEZ-Lv105 lentivirus vectors.The sequences of the recombinant plasmids were confirmed correctly by PCR and DNA sequence.The enhanced green fluorescent protein (eGFP) could be observed after recombinant lentiviruses were cotransfected into 293T cells.Conclusions The TNFAIP1 overexpressed lentivirus vector is successfully constructed,which provides a molecular tool for further study of TNFAIP1 gene in optic nerve glioma.