Construction and expression in Escherichia coli of hybrid genes composed of sequences encoding diphtheria toxin and human CD4 receptor

Derivatives of natural toxins possessing substituted receptor-recognition domains of different specificities can be used as instruments for the selective elimination of target cells. We have constructed two different types of hybrid genes that encode proteins composed of diphtheria toxin (DT) lacking the C-terminal residues that mediate toxin binding fused with the N-terminal region of human CD4 (Lys 10 to Glu 152). One of these hybrids encodes a protein with CD4 at the N terminus, while the other encodes a protein with CD4 at the C terminus. The stability of these two proteins was dramatically different. We could not detect a full-size product when the first construct was expressed in Escherichia coli. In contrast, proteins encoded by the second construct were more stable. In the latter case, the amount of full-size hybrid protein was 1–2% of the total cell protein. We speculate on the involvement of the region that resembles the processing site of Pseudomonas aeruginosa exotoxin A in the proteolytic degradation of the product encoded by the first type of hybrid.