Cloning and expression of mouse HMGB1 B box coding sequence

AIM: To clone and express mouse High mobility group box 1 (HMGB1) B box coding sequence. METHODS: Total RNA was extracted from the lung tissue of a newborn mouse and the HMGB1 B box coding sequence was obtained by RT-PCR using specific primers. The B box coding sequence was cloned into BamHI and EcoRI site of pGEX-4T-2 expression vector and confirmed by sequencing. After transforming E.coli BL21(DE3), the recombinant bacteria was induced at 30℃ for 5 h, the fusion protein GST-B box was analyzed by SDS-PAGE. RESULTS: DNA sequencing results showed that the mouse HMGB1 B box coding sequence was exactly consistent with the sequence reported in GenBank, and the SDS-PAGE analysis demonstrated that HMGB1 B box protein was expressed in E.coli BL21 (DE3). The protein band amounted to 30% of total bacteria protein. CONCLUSION: Mouse HMGB1 B box coding sequence is successfully cloned and expressed in E.coli.