Construction and regulated expression of mouse TGF-β1 recombinant adenovirus vector

AIM: To construct a recombinant adenovirus vector which regulated expression of mouse transforming growth factor beta (TGF-β1). METHODS: Total RNA was extracted from Balb/c mice spleen pre-treated with Con A to clone TGF-β1. pTRE-shuttle vector was used as mediator to ligate TGF-β1 gene and the backbone of the replication-incompetent adenoviral vector. The constructed recombinant adenovirus contains tetracycline-responsive element which can regulate the expression of inserted genes. Identifying the desired recombinant adenovirus, it was packaged in HEK 293 cells. Supernatant of high titer adenovirus was collected to detect the TGF-β1 gene expression by ELISA kit. RESULTS: The constructed recombinant adenovirus had been identified by restriction endonucleases cutting, sequencing and PCR. In the supernatant of infected cells, it expressed the mice TGF-β1 in high concentration to 91.16 ng/L. CONCLUSION: The mice TGF-β1 recombinant adenovirus has been built which make a basis for further research and use in gene therapy.