What factors contribute to the low expression of some proteins?
Successful expression and purification of a target protein in an IMPACT vector depend on the following factors: (a) E. coli strain; (b) culture conditions (e.g., temperature, aeration, cell density); (c) induction and protein expression conditions (temperature, time, IPTG concentration); (d) expression level of the fusion protein; (e) solubility of the fusion protein; (f) efficiency of thiol-induced cleavage; (g) solubility of the target protein after the cleavage. The expression of the fusion protein in the C-terminal fusion vectors (pTXB1,3,pTYB1,2,3 and 4) may be primarily determined by the expression level of the N-terminal target protein. In the case of the N-terminal fusion vectors (pTYB21, pTYB22, pTYB11 and 12) the expression level is less dependent on the target protein. It is conceivable that different target proteins, due to certain structural constraints, may prefer either C-terminal or N-terminal fusion to allow proper folding of the fusion precursor and a high level of pr
Successful expression and purification of a target protein in an IMPACT vector depend on the following factors: (a) E. coli strain; (b) culture conditions (e.g., temperature, aeration, cell density); (c) induction and protein expression conditions (temperature, time, IPTG concentration); (d) expression level of the fusion protein; (e) solubility of the fusion protein; (f) efficiency of thiol-induced cleavage; (g) solubility of the target protein after the cleavage. The expression of the fusion protein in the C-terminal fusion vectors (pTXB1,3,pTYB1,2,3 and 4) may be primarily determined by the expression level of the N-terminal target protein. In the case of the N-terminal fusion vectors (pTYB21, pTYB22, pTYB11 and 12) the expression level is less dependent on the target protein. It is conceivable that different target proteins, due to certain structural constraints, may prefer either C-terminal or N-terminal fusion to allow proper folding of the fusion precursor and a high level of pr
