[Expression of Epstein-Barr virus latent membrane protein 2 in murine fibroblasts by retroviral-mediated gene transfer].
OBJECTIVE
This study is to find an efficient way to express EB virus latent membrane protein 2 in mammalian cells, and provide for further investigate on the function of LMP2 protein and its role in cellular immunity.


METHODS
The LMP2 gene was cloned into the EcoR I site of a retroviral vector LXSN and the recombinant LXSN-LMP2 was transfected into PT67 by lipofect TAMINE. After screened by G418, the supernatant of G418-resistant cells was used to infect murine fibroblasts NIH 3T3 to determine the virus titer. DNA was extracted from transfected cells and tested by PCR. Indirect immunofluorescence assay was used to detect the expression of the inserted gene.


RESULTS
The virus titer was 5.8 x 10(5) PFU/ml. The result of PCR showed that the LMP2 gene had been integrated into the DNA of transfected cells. Indirect immunofluorescence showed that the LMP2 gene had been expressed in the murine fibroblasts.


CONCLUSIONS
EB virus LMP2 gene had been integrated into the genome of cells by retroviral-mediated transfer and the target gene had been expressed.
Zhu
W
W
Institute of Virology, Chinese Academy of Preventive Medicine, Beijing 100052, China.
Zhou
L
L
Yao
J
J
D016475:3T3 Cells; D000818:Animals; D002478:Cells, Cultured; D015870:Gene Expression; D005822:Genetic Vectors; D004854:Herpesvirus 4, Human; D051379:Mice; D012190:Retroviridae / Q000235:genetics*; D014162:Transfection; D014763:Viral Matrix Proteins / Q000096:biosynthesis* / Q000235:genetics
C037662:EBV-associated membrane antigen, Epstein-Barr virus; D014763:Viral Matrix Proteins