Purpose: To construct a recombinant lentiviral vector for human amelogenin and to investigate the amelogenin expression of recombinant lentivirus FUAmW in 293T cell line.
Methods: A lentiviral expression vector for human amelogenin was constructed by recombinant DNA technique. Recombinant plasmid FUAmW was confirmed by restriction endonuclease and DAN sequence analysis respectively. High tites recombinant lentivirus were prepared from 293T cells by polytheylenimine(PEI) mediated transient cotransfection. The generated recombinant FUGW viruses and recombinant FUAmW viruses were used to infect 293T cells, respectively. The expression of human amelogenin in 293T cells was detected by RT-RCR and Western-blot.
Results: The sequence analysis of recombinant plasmid FUAmW showed that the human amelogenin encoding mature protein was inserted into lentiviral vector FUW accurately. Human amelogenin expressions were observed in 293T cells 72 hours after infecting with recombinant FUAmW viruses.
Conclusions: The recombinant lentiviral vector FUAmW can be constructed correctly and transfected into 293T cells. Human amelogenin can be expressed in 293T cells infected with recombinant FUAmw virus. Supported by National Natural Science Foundation of China (Grant No.30672315).