[Prokaryotic expression and purification of recombinant rat HMGB1-A].

Objective To obtain the recombinant A-domain of high mobility group box 1 (HMGB1-A) protein. Methods Using genetic engineering techniques, we cloned and recombined rat HMGB1-A gene. Using prokaryotic expression technique, we expressed and purified the recombinant rat HMGB1-A protein, which was verified by Western blotting. Results Agarose gel electrophoresis analysis showed that HMGB1-A cassette gene size was about 250 bp; double digestion for identifying the recombinant clone pUC-A showed that the product size was about 3000 bp, 250 bp; PCR for identifying the recombinant clone pGEX-A showed that the product size was about 250 bp. SDS-PAGE revealed that the product size was about 36 000, which was consistent with the expectation. Conclusion The recombinant rat HMGB1-A was successfully expressed and purified.