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CJAP ›› 2019, Vol. 35 ›› Issue (6): 573-576.doi: 10.12047/j.cjap.5856.2019.126

• TECHNICAL NOTES • Previous Articles    

Construction of eukaryotic expression plasmid of pcDNA3.1(+)- CTGF and its expression in human osteoblast-like cells SaOS-2

MA Ke-feng, YANG Shu-guang, LIU Shao-jun   

  1. Institute of Military Cognition and Brain Sciences, Academy of Military Medical Sciences, Academy of Military Sciences, Beijing 100850, China
  • Received:2019-03-29 Online:2019-11-28 Published:2020-04-02

Abstract: Objective: To construct pcDNA3.1(+) eukaryotic expression plasmid of connective tissue growth factor(CTGF), and detected its expression in human osteoblast-like cells SaOS-2, which provides a technical support for further research on the mechanism of CTGF gene in bone development and bone repair process. Methods: The whole sequence of CTGF gene was cloned in vitro by polymerase chain reaction(PCR) method and connected to the linear pcDNA3.1(+) vector for constructing pcDNA3.1(+)-CTGF eukaryotic expression plasmid by homologous recombination technology. The plasmid was identified by sequencing. After identification, it was transfected into SaOS-2 cells and its expression was detected at 48 h. Results: pcDNA3.1(+)-CTGF eukaryotic expression recombinant plasmid was successfully constructed, which was confirmed by sequencing. Compared with the control group, CTGF expression level was significantly up-regulated after transfection of SaOS-2 cells for 48 h, up to five times as much as the control group. Conclusion: pcDNA3.1(+)-CTGF eukaryotic expression plasmid was successfully constructed and could be stably expressed in human osteoblasts-like cell SaOS-2, which laid a foundation for further study on the regulatory mechanism of CTGF gene on bone formation.

Key words: connective tissue growth factor(CTGFAT), human osteoblast-like cells, plasmid construction, eukaryotic expression, bone formation

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