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Expression, secretion and processing of hirudin in E. coli using the alkaline phosphatase signal sequence
Author(s) -
Dodt Johannes,
Schmitz Thomas,
Schäfer Thomas,
Bergmann Cornelia
Publication year - 1986
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(86)80721-9
Subject(s) - periplasmic space , signal peptide , microbiology and biotechnology , hirudin , alkaline phosphatase , coding region , biochemistry , gene , secretion , biology , escherichia coli , chemistry , peptide sequence , thrombin , enzyme , platelet , immunology
A DNA fragment coding for the E. coli phoA signal peptide was synthesized and inserted into the expression vector pKK223‐3. A single Hin dIII restriction site is located just at the end of the signal sequence. A gene coding for the proteinase inhibitor hirudin, which has previously been synthesized, was inserted into this Hin dIII site. The hybrid protein was expressed under control of the tac‐promoter and secreted into the periplasm of E. coli . From the periplasmic fraction two processed proteins were isolated. One of these was identical with desulfatohirudin and also had similar biological properties.