Expression of human flt3 ligand and thrombopoietin genes in a bone marrow stromal cell line by internal ribosome entry site (IRES)sequence
Författare
Summary, in English
Objective To explore the feasibility of retroviral-mediated Flt 3 ligand (FL) and thrombopoietin (Tpo) genes transferred into and expressed in a bone marrow stromal cell line HFCL by internal ribosome entry site（IRES）sequence. Methods IRES sequence, FL and Tpo cDNA were recombined with retroviral vector pLXSN by gene recombination technology. The recombinant plasmid was transferred into retrovirus packaging cell line PA317 by lipofectamine, and the resistant clones were selected by G418 selective medium. mRNA expression in HFCL cells and integration of genome DNA were assayed by RT-PCR and genomic DNA PCR. The biological activities of FL and Tpo in the culture were investigated by CFU-GM assay and Tpo dependent cell line TD-3,respectively. Results The recombinant plasmid pLFTSN was successfully constructed. In the genome of these transfected target cells, Neo gene and FL and Tpo cDNA were intergrated, the expression of FL and Tpo mRNA was detected in HFCL cells. The specific activities of FL and Tpo in the culture indicated that HFCL cells transfected with FL and Tpo cDNA could significantly express FL and Tpo in vitro. Conclusion FL gene and Tpo gene were simultaneously expressed in bone marrow stromal cell line by the regulation of IRES sequence. These results provide a basis for studies on hematopoietic regulation by gene transfected bone marrow stromal cells.
Publiceringsår
1999
Språk
Kinesiska
Sidor
624-627
Publikation/Tidskrift/Serie
Chinese Journal of Hematology
Volym
20
Issue
12
Länkar
Dokumenttyp
Artikel i tidskrift
Förlag
Zhongguo yi xue ke xue yuan
Ämne
- Medicinal Chemistry
Nyckelord
- Platelet-derived growth factor
- Ligands
- Cell line，HFCL
- Gene
- Gene expression
Status
Published
ISBN/ISSN/Övrigt
- ISSN: 0253-2727