Cloning of the Epstein-Barr Virus Gene BNLF-1 and Its Expression in PA317 Cells.
We have constructed the expression vector pZipNexSV (x)-LMP by the recombinant DNA technology. The oncogene BNLF-1 of Epstein-Barr virus was inserted into retroviral vector pZip-NexSV (x) subjecting the BNLF-1 under the control of both promoter/enhancer sequence of MLV-LTR and EDL1 promoter of BNLF-1 gene itself. The fusion gene DNA was transfected into PA317 cells by electroporation. Ten of 54 G418-resistant colonies were allowed to grow up and the DNAs were checked by PCR. The results showed that 6 colonies carried the BNLF-1 gene. Northern hybridization and Western blotting to detect the transcription and translation of the BNLF-1 gene demonstrated that the fusion gene was expressed efficiently under the control of EDL1 promoter in PA317 cells.
Ma
Xian-Yong
XY
Laboratory of Tumor Molecular Biology, Cancer Research Institute, Human Medical University, Changsha 410078, China.
Yao
Kai-Tai
KT