[Establishment and identification of mouse lymphoma cell line EL4 expressing red fluorescent protein].
This study was purposed to construct a lentiviral vector encoding red fluorescent protein (DsRed) and transfect DsRed into EL4 cells for establishing mouse leukemia/lymphoma model expressing DsRed. The bicistronic SIN lentiviral transfer plasmid containing the genes encoding neo and internal ribosomal entry site-red fluorescent protein (IRES-DsRed) was constructed. Human embryonic kidney 293FT cells were co-transfected with the three plasmids by liposome method. The viral particles were collected and used to transfect EL4 cells, then the cells were selected by G418. The results showed that the plasmid pXZ208-neo-IRES-DsRed was constructed successfully, and the viral titer reached to 10(6) U/ml. EL4 cells were transfected by the viral solution efficiently. The transfected EL4 cells expressing DsRed survived in the final concentration 600 microg/ml of G418. The expression of DsRed in the transfected EL4 cells was demonstrated by fluorescence microscopy and flow cytometry. In conclusion, the EL4/DsRed cell line was established successfully.
Laboratory of Transplantation Immunology, Xuzhou Medical College, Xuzhou 221002, Jiangsu Province, China.
Yan-Jie
YJ
Li
Jiang
J
Cao
Chong
C
Chen
Dong-Yang
DY
Wang
Ling-Yu
LY
Zeng
Xiu-Ying
XY
Pan
Kai-Lin
KL
Xu
D000818:Animals; D045744:Cell Line, Tumor; D005434:Flow Cytometry; D005822:Genetic Vectors; D016086:Lentivirus / Q000235:genetics*; D008164:Luminescent Proteins / Q000235:genetics*; D008223:Lymphoma / Q000235:genetics*; D051379:Mice; D008810:Mice, Inbred C57BL; D014162:Transfection; D000097573:Red Fluorescent Protein
D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein