Expression of chemically synthesized -neo-endorphin gene fused to E. coli alkaline phosphatase

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Abstract
An α-neo-endorphin (αNE) gene, which we previously synthesized chemically and inserted into E.coli (β-galactosidase gene of pK013 plasmid, has been excised and fused to E.coli alkaline phosphatase (APase) gene. One of the transformants was named E15/pAαNEl. Under the APase gene regulation, APase-αNE chimeric protein was expressed at 1.3 × 106 molecules per cell, and accounted for about 60+ of total cellular proteins. The HPLC pattern of CNBr treated E15/pAαNEl was very simple reflecting the high content of the chimeric protein and low numbers of methionine residues in it. A series of genes encoding APase-αNE chimeric proteins in which 30 to 94 C-terminal amino acid residues were replaced by (met)-αNE, was cloned in E.coli. Transportation of the chimeric proteins to periplasmic space was studied. All chimeric proteins were apparently processed by signal peptidase but few, if any, was transported to the periplasmic space.