[High expression of human thrombopoietin mutant fusion protein in E. coli].

Objective: To obtain human thrombopoietin (Tpo)mutant cDNA encoding mature peptide N terminal 1 approximately 196 amino acids and investigate its expression in E. coli JM109.

Methods: Polymerase chain reaction and DNA recombination techniques were employed. PCR product was inserted into pUC19 vector and sequenced and then cloned into expression vector pMAL-c2.

Results: E. coli JM109 cells with plasmid pMAL -MBP/TpoM were induced by IPTG for 4-5 hours. SDS-PAGE analysis showed that MBP/TpoM molecular weight is about 63KD. Scanning analysis indicated that expressed protein accounts up to 37% of total E. coli proteins.

Conclusion: The Tpo mutant of interest is successfully expressed in E. coli JM109 cells.