Open AccessHigh-level production of biologically active human alpha 1-antitrypsin in Escherichia coli.
Author(s) -
Michael Courtney,
Anne Buchwalder,
Luc-Henri Tessier,
Michael Jaye,
Annie Benavente,
Alain Balland,
Vijay Kohli,
Richard Lathe,
Paul Tolstoshev,
JeanPierre Lecocq
Publication year - 1984
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.81.3.669
Subject(s) - escherichia coli , complementary dna , biology , microbiology and biotechnology , gene , protein biosynthesis , ribosome , biochemistry , rna
A cDNA clone containing the complete human alpha 1-antitrypsin sequence was isolated from a human liver cDNA bank by screening with a chemically synthesized oligonucleotide probe. DNA sequences encoding the alpha 1-antitrypsin mature polypeptide were inserted into an Escherichia coli expression vector that allows transcription from the efficient leftward promoter of bacteriophage lambda (PL) and initiation of translation at the lambda cII gene ribosome-binding site. This construction resulted in the induction of a 45-kilodalton protein at a level of approximately 15% of total cell protein. The polypeptide produced was recognized by antisera raised against human alpha 1-antitrypsin protein and displayed normal biological activity in an in vitro antielastase assay.