Establishment of mouse SP2/0 cell line stably expressing bcr-abl fusion gene fragment.
- Author:
Yang-Wen JIANG
1
;
Li QIAN
;
Wei LIU
;
Wei-Juan GONG
;
Bing WANG
;
Jun GUAN
;
Ming-Chun JI
Author Information
1. Department of Hematology, Affiliated Hospital of Yangzhou University, Yangzhou 225001, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Cell Line;
Cell Line, Tumor;
Fusion Proteins, bcr-abl;
genetics;
Gene Expression Regulation, Neoplastic;
Genetic Vectors;
genetics;
Humans;
K562 Cells;
Mice;
Mice, Inbred BALB C;
Multiple Myeloma;
genetics;
pathology;
NIH 3T3 Cells;
Peptide Fragments;
genetics;
RNA, Messenger;
biosynthesis;
genetics;
Retroviridae;
genetics;
Reverse Transcriptase Polymerase Chain Reaction;
Transfection;
methods
- From:
Journal of Experimental Hematology
2005;13(4):601-604
- CountryChina
- Language:Chinese
-
Abstract:
To establish SP2/0 cell line H-2(d) stably expressing bcr-abl fusion gene fragment, the bcr-abl fusion gene was subcloned into retroviral vector pLXSN from pGEMbcr-abl. The recombinant retroviral vector pLXSNbcr-abl was transfected into PT67 packaging cells with the help of lipofectamine. The positive clones were selected out and cultured after G418 selection. Then viral supernatant was collected to determine viral titer, the viral titer was 2 x 10(7) CFU/ml. The SP2/0 cells were infected with the collected viral supernatant. The results showed that after G418 selection, the bcr-abl fusion gene was integrated into the chromosome of SP2/0 cells infected stably, with recombinant retrovirus and expressed in SP2/0 cells confirmed by PCR and RT-PCR respectively. In conclusion, the mouse tumor cell lines expressing the bcr-abl fusion protein were successfully established and would be used as a experimental cell model for anti-CML immunotherapy.
