Construction and identification of the recombinant adenovirus shuttle vector carrying the human Toll-like receptor 2 extracellular domain gene

Objective:To clone the human TLR2 extracellular domain gene and construct the recombinant adenovirus shuttle plasmid pAdTrack-CMV-TLR2. Methods: The TLR2 extracellular domain gene was amplified from total RNA of human peripheral blood mononuclear cells by RT-PCR and inserted into the pUCm-T vector. The recombinant was transformed into E coli DH5α. After confirmed by enzyme digestion and sequencing, the DNA fragment, digested with KpnⅠand Hind Ⅲ, was directionally cloned into the adenovirus shuttle plasmid pAdTrack-CMV. The recombinant plasmid was verified by double digestion and DNA sequencing. Results: The gene fragment of 1000bp was obtained by RT-PCR. After TA cloned, its sequence conformed with that reported in the GenBank, and the cloned gene was exactly inserted into the adenovirus shuttle vector. Conclusion: The TLR2 extracellular domain gene was correctly cloned and the recombinant adenovirus shuttle vector successfully constructed.