Construction of venus vector carrying IGFBP7 gene and its expression in K562 cells.
- Author:
Shui-Yan WU
1
;
Shao-Yan HU
;
Jian-Nong CEN
;
Zi-Xing CHEN
Author Information
1. Department of Hematology, Children's Hospital of Soochow University, Suzhou, Jiangsu Province, China.
- Publication Type:Journal Article
- MeSH:
Cloning, Molecular;
Gene Expression;
Genetic Vectors;
Humans;
Insulin-Like Growth Factor Binding Proteins;
genetics;
K562 Cells;
Lentivirus;
genetics;
Plasmids;
Transfection
- From:
Journal of Experimental Hematology
2012;20(1):164-167
- CountryChina
- Language:Chinese
-
Abstract:
The aim of this study was to construct venus vector carrying the gene encoding insulin-like growth factor binding protein 7 (IGFBP7), which provides an effective platform for exploring the function of this gene in leukemia. After digestion by restriction endonuclease, the IGFBP7 gene was recombined with the transfer plasmid. The venus particles were packaged using 293T cells to transfect K562 cells, and identification was performed by means of flow cytometry, RT-PCR and Western blot. The results showed that the sequence of cloned IGFBP7 gene was the same as that in GenBank. The size of product restricted by BamHI was same as the predicted one. GFP expression was observed in 293T and K562 cells with the fluorescent microscopy and flow cytometry. The expression level of mRNA and protein of IGFBP7 was confirmed by RT-PCR and Western blotting in K562 cells. It is concluded that venus vector carrying IGFBP7 gene has been successfully constructed and provides basis for exploring function of IGFBP7 in K562 cells.
