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Novel strategy for protein production using a peptide tag derived from Bacillus thuringiensis Cry4Aa
Author(s) -
Hayakawa Tohru,
Sato Shinya,
Iwamoto Shigehisa,
Sudo Shigeo,
Sakamoto Yoshiki,
Yamashita Takaaki,
Uchida Motoaki,
Matsushima Kenji,
Kashino Yohko,
Sakai Hiroshi
Publication year - 2010
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/j.1742-4658.2010.07704.x
Subject(s) - bacillus thuringiensis , escherichia coli , peptide , recombinant dna , fusion protein , biology , inclusion bodies , treponema , antigen , bacteria , biochemistry , microbiology and biotechnology , chemistry , virology , gene , syphilis , genetics , human immunodeficiency virus (hiv)
Numerous proteins cannot be sufficiently prepared by ordinary recombinant DNA techniques because they are unstable or have deleterious effects on the host cell. One idea to prepare such proteins is to produce them as protein inclusions. Here we developed a novel system to effectively prepare proteins by using peptide tags derived from the insecticidal Cry toxin of a soil bacterium, Bacillus thuringiensis . Fusion with this peptide tag, designated 4AaCter, facilitates the formation of protein inclusions of glutathione S ‐transferase in Escherichia coli without losing the enzyme activity. Application of 4AaCter to the production of syphilis antigens TpN15, TpN17 and TpN47 from Treponema pallidum yielded excellent results, including a dramatic increase in the production level, simplification of the product purification and high reactivity with syphilis antibody. The use of 4AaCter may provide an innovational strategy for the efficient production of proteins.