tailieunhanh - Báo cáo khoa học: Novel strategy for protein production using a peptide tag derived from Bacillus thuringiensis Cry4Aa

Numerous proteins cannot be sufficiently prepared by ordinary recombi-nant 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. | ỊFEBS Journal Novel strategy for protein production using a peptide tag derived from Bacillus thuringiensis Cry4Aa Tohru Hayakawa1 Shinya Sato1 2 Shigehisa Iwamoto2 Shigeo Sudo2 Yoshiki Sakamoto1 Takaaki Yamashita1 Motoaki Uchida1 Kenji Matsushima1 Yohko Kashino1 and Hiroshi Sakai1 1 Graduate Schoolof NaturalScience and Technology Okayama University Japan 2 Department of Bioscience Japan Lamb Co. Ltd Development Division Okayama Japan Keywords 4AaCter peptide tag Bacillus thuringiensis Cry4Aa Escherichia coli TpN syphilis antigen Correspondence H. Sakai Graduate Schoolof Natural Science and Technology Okayama University 3-1-1 Tsushima-Naka Okayama 700-8530 Japan Fax Tel 81 86 251 8203 E-mail sakahrsh@ Database The nucleotide sequence of the synthetic gene cry4Aa-S2 is available in the EMBL GenBank DDBJ databases under accession number AB513706 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. Received 26 February 2010 revised 11 April 2010 accepted 3 May 2010 doi Introduction Cry toxin a specific insecticidal .