Optimization of condition for prokaryotic expression of recombinant tetanus toxin heavy chain fragment C protein / 中国生物制品学杂志

ABSTRACT

@#Objective To optimize the condition for prokaryotic expression of recombinant tetanus toxin heavy chain fragment C(rTTHc) protein.Methods The rTTHc gene fragment after optimization of codon was inserted into prokaryotic expression vector pET30a,low temperature expression vector pCold Ⅱ and high temperature expression vector pBV220separately.The constructed recombinant plasmids were transformed to E.coli BL21(DE3).The expression levels and solubility of recombinant protein at various temperatures were compared.Results The expression level of pBV220-rTTHc after induction at 42 ℃ for 4 h was relatively low,and the protein solubility was poor.The expression level of pET30a-rTTHc after induction at 37 ℃ for 4 h was equivalent to that of pCold-rTTHc after induction at 15 ℃ for 8 h,while the solubility of the former was slightly lower than that of the latter.However,both the expression level and solubility of pET30a-rTTHc after induction at 28 ℃ for 4 h were high,while the expression time was short.Conclusion The pET30a-rTTHc induced by2 mg/mL IPTG at 28℃ is optimal for high expression of rTTHc protein.