title: Effect of Glycine and Triton X-100 on secretion and expression of ZZ-EGFP fusion protein.
abstract: Two-factor and three-level fractional factorial design was employed for evaluation of the effect of Glycine and Triton X-100 on the secretion and expression of ZZ-EGFP fusion proteins. Varying contents of glycine (0%, 1%, 2%) and Triton X-100 (0%, 1%, 2%) were added into shaking flasks, respectively, and supplied with appropriate volume of ampicillin (total 9 combinations; group at concentration zero serving as control) to promote more ZZ-EGFP diffuse into liquid culture medium. Fluorescent intensity in the culture supernatant was detected. A standard curve could be generated on the basis of fluorescent intensity and protein concentration. The expression level of ZZ-EGFP fusion proteins was estimated by checking the protein standard curve concentration fluorescene intensity. Results show that when the culture medium contains 2% Glycine and 1% Triton X-100, the expression level of ZZ-EGFP was able to be greatly increased. Further experiments revealed that absorbance value (A600) in the experiment group, whose culture medium contains 2% Glycine and 2% Triton X-100, is significantly lower than other groups in the present experiment. These results indicate that the culture medium containing appropriate quantity of Glycine and Triton X-100 is favourable to the secretion and expression level of ZZ-EGFP in gene-engineering bacteria Escherichia coli HB101.
authors.affiliation: Department of Basic medicine, Weifang Medical University, WeiFang 261042, PR China.
authors.forename: Jin-Bao
authors.initials: JB
authors.lastname: Tang
authors.affiliation: Department of Basic medicine, Weifang Medical University, WeiFang 261042, PR China.
authors.forename: Hong-Ming
authors.initials: HM
authors.lastname: Yang
authors.affiliation: Weihai International Biotechnology R&D Center, Shandong University, WeiHai 262409, PR China.
authors.forename: Shu-Liang
authors.initials: SL
authors.lastname: Song
authors.affiliation: Weihai International Biotechnology R&D Center, Shandong University, WeiHai 262409, PR China.
authors.forename: Peng
authors.initials: P
authors.lastname: Zhu
authors.affiliation: Weihai International Biotechnology R&D Center, Shandong University, WeiHai 262409, PR China. Electronic address: jiaiguo@sdu.edu.cn.
authors.forename: Ai-Guo
authors.initials: AG
authors.lastname: Ji
keywords: E. coli HB101; Gene-engineering bacteria; Glycine; Secretion; Triton X-100; ZZ–EGFP