title: Cloning and Expression of Tetanus Toxin Fragment C in E.coli.
abstract: The fragment C of tetanus toxin was amplified from Clostridium tetani DNA by PCR. This fragment was cloned into expression vector pET-28a(+),under the control of the T7 promoter. Expression of this plasmid in E.coli resulted in the production of a protein consisting of 6xHis of the vector fused to the N-terminal 451 amino acids of tetanus toxin. After induction with 1 mmol/L IPTG, TTC was expressed in E.coli BL21(DE3). The protein product accounted for 8.2% of the bacteria total protein in soluble form, SDS-PAGE and Western blot analysis of TTC recombinant protein confirmed this result. The expression products were also purified by Ni(2+)-IDA-Sephrose 6B column. Immunization of mice with rTTC resulted in the production of antibodies that were able to protect mice against a challenge with tetanus toxin furthermore, rTTC in vivo appeared to be able to undergo retrograde axonal transport.
authors.affiliation: Shanghai Institute of Biochemistry, the Chinese Academy of Sciences, Shanghai 200031, China. xfwu@sunm.shcnc.ac.cn
authors.forename: Hua-Jun
authors.initials: HJ
authors.lastname: He
authors.forename: Zhi-Yong
authors.initials: ZY
authors.lastname: He
authors.forename: Hui-Juan
authors.initials: HJ
authors.lastname: Shi
authors.forename: Wei
authors.initials: W
authors.lastname: Zhu
authors.forename: Guan-Zhen
authors.initials: GZ
authors.lastname: Yang
authors.forename: Qin-Sheng
authors.initials: QS
authors.lastname: Yuan
authors.forename: Xiang-Fu
authors.initials: XF
authors.lastname: Wu