Cloning of NPC CNE1 cell strain EBV-LMP1 CDNA and the research of cell outside stability of LMP1

Objective:To clone CNE1 cell strain EBV-LMP1 cDNA,transfect into plasmid vector and detect CNE1 cell strain LMP1 expression outside cell stability.Methods: Immunohistochemical method was used to detect CNE1 LMP（+） cell expression of LMP1.Amplified EBV-LMP1 cDNA from CNE cell by RT-PCR technique was cloned into pGEM-T easy vector and sequenced.The constructed recombinant plasmid was transferred into E.coli JM109.and indentified by restriction analysis and eletrophrasis.Sequence including 3 sections LMP1 sequence out of CNE1 cell strain were amplified and sequenced.Results: LMP1 protein was expressed highly in the cell,and the result of sequence had some variations compared with the known sequence.The expression of 3 sections LMP1 sequence out of CNE1 cell strain was stable.Conclusions: The LMP1 gene is successfully amplified and cloned into plasmid vector.The stable expression of LMP1 out of CNE1 cell provides the basic pathogenesis for studying the oncogenic role of LMP1 and further study of antibody function using in comfirmation of NPC radiotherapy target.