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Expression of human flt3 ligand and thrombopoietin genes in a bone marrow stromal cell line by internal ribosome entry site (IRES)sequence

Zhang, Yi ; Tang, Pei-Hsien ; Li, Xiusen ; Jiang, Feizi ; Sun, Jianmin LU and Mao, Ning (1999) In Chinese Journal of Hematology 20(12). p.624-627
Abstract
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.... (More)
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. (Less)
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author
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publishing date
type
Contribution to journal
publication status
published
subject
keywords
Platelet-derived growth factor, Ligands, Cell line，HFCL, Gene, Gene expression
in
Chinese Journal of Hematology
volume
20
issue
12
pages
624 - 627
publisher
Zhongguo yi xue ke xue yuan
external identifiers
  • scopus:0033302157
ISSN
0253-2727
language
Chinese
LU publication?
no
additional info
The information about affiliations in this record was updated in December 2015. The record was previously connected to the following departments: Experimental Clinical Chemistry (013016010)
id
17783455-0a87-4f3e-9008-47d535f2a0d4 (old id 1116088)
alternative location
http://www.wanfangdata.com.cn/qikan/periodical.articles/zhxyx/zhxy99/zhxy9912/991202.htm
date added to LUP
2016-04-01 15:53:29
date last changed
2022-01-28 07:49:37
@article{17783455-0a87-4f3e-9008-47d535f2a0d4,
  abstract     = {{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.}},
  author       = {{Zhang, Yi and Tang, Pei-Hsien and Li, Xiusen and Jiang, Feizi and Sun, Jianmin and Mao, Ning}},
  issn         = {{0253-2727}},
  keywords     = {{Platelet-derived growth factor; Ligands; Cell line，HFCL; Gene; Gene expression}},
  language     = {{chi}},
  number       = {{12}},
  pages        = {{624--627}},
  publisher    = {{Zhongguo yi xue ke xue yuan}},
  series       = {{Chinese Journal of Hematology}},
  title        = {{Expression of human flt3 ligand and thrombopoietin genes in a bone marrow stromal cell line by internal ribosome entry site (IRES)sequence}},
  url          = {{http://www.wanfangdata.com.cn/qikan/periodical.articles/zhxyx/zhxy99/zhxy9912/991202.htm}},
  volume       = {{20}},
  year         = {{1999}},
}