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High level expression of soluble angiogenin in Escherichia coli
Author(s) -
Yoon Jong Myung,
Kim Seung Ho,
Kwon Oh Byung,
Han Seung Hee,
Kim Byong Kak
Publication year - 1999
Publication title -
iubmb life
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.132
H-Index - 113
eISSN - 1521-6551
pISSN - 1521-6543
DOI - 10.1080/15216549900201283
Subject(s) - angiogenin , affinity chromatography , escherichia coli , periplasmic space , biochemistry , chemistry , signal peptide , microbiology and biotechnology , ion chromatography , sepharose , reticulocyte , biology , gene , messenger rna , recombinant dna , enzyme , angiogenesis , cancer research
Human angiogenin was genetically engineered and contained the E. coli Omp A signal sequence for secreting soluble angiogenin to the periplasm under tac promoter control. The angiogenin sequence was encoded in a single gene and expressed as a 14.4 kilodalton soluble protein in E. coli. It was purified by CM‐Sepharose ion‐exchange chromatography and by a heparin‐Sepharose affinity chromatography procedure. The biological activity of angiogenin was established by its ability to inhibit mRNA‐dependent rabbit reticulocyte cell‐free translation.