Efficient production of biologically active human prolactin in Escherichia coli.

摘要: Abstract To obtain an adequate amount of human prolactin (hPRL) for elucidation the structure-function relationship, we have expressed hPRL cDNA in Escherichia coli (E. coli) by using a high-expression vector. The vector contained chimeric gene encoding fusion protein A, peptide sensitive to collagenase digestion and hPRL, which was inserted downstream right direction promotor λ phage. resulting purified through three column chromatographies immunoglobulin G-linked Sepharose 4B, DEAE-5PW, phenyl-5PW. In typical experiment, final sample with purity more than 80% obtained recovery 40% judged enzyme-linked immunosorbent assay (ELISA). thus digested collagenase, reactive anti-hPRL antibody phenyl-5PW chromatography. found be identical authentic respect amino acid composition N-terminal sequence 20 residues, except that it additional four acids at end. This presumed derived from collagenase-target sequence. as active hormone either immunologically ELISA or biologically growth stimulatory effect on rat Nb2 lymphoma cells.