title: High-efficiency expression of TAT-bFGF fusion protein in Escherichia coli and the effect on hypertrophic scar tissue.
abstract: BACKGROUND
  Basic fibroblast growth factor (bFGF) is a member of the fibroblast growth factor family that has effects on wounding healing and neuro-protection. However, it is difficult to use bFGF to treat diseases that are separated by physiological barriers, such as the dermal barrier and blood brain barrier.
  
  
  RESULTS
  To improve bFGF's penetration ability, we fused the recombinant human fibroblast growth factor (rhbFGF) gene with TAT. We constructed a pET3c vector that contained the recombinant bFGF gene and successfully expressed this gene in the E. coli strain BL21 (DE3) pLsS. The fusion protein was purified using CM Sepharose FF and heparin affinity chromatography. The purity of the TAT-rhbFGF was greater than 95%, as detected by SDS-PAGE. An in vitro MTT trial revealed that the modified bFGF significantly promoted the proliferation of NIH3T3 cells. The cell penetration trial and the mouse skin penetration trial demonstrated that the fusion protein had certain penetration abilities. The animal experiments confirmed that TAT-rhbFGF was effective in the treatment of the hypertrophic scars.
  
  
  CONCLUSIONS
  We have successfully expressed and purified a TAT-rhbFGF fusion protein in this study. Our results have shown that the fusion protein had a greater ability to penetrate the dermal skin layer. TAT-rhbFGF improved the physical appearance of hypertrophic scars. TAT-rhbFGF may be a potential fusion protein in the treatment of dermal disorders, including hypertrophic scar.
authors.affiliation: Key Laboratory Biotechnology Pharmaceutical Engineering, Wenzhou Medical University, Chashan University Park, Wenzhou, 325035, China.
authors.forename: Xuechao
authors.initials: X
authors.lastname: Jia
authors.affiliation: Key Laboratory Biotechnology Pharmaceutical Engineering, Wenzhou Medical University, Chashan University Park, Wenzhou, 325035, China.
authors.forename: Haishan
authors.initials: H
authors.lastname: Tian
authors.affiliation: Key Laboratory Biotechnology Pharmaceutical Engineering, Wenzhou Medical University, Chashan University Park, Wenzhou, 325035, China.
authors.forename: Lu
authors.initials: L
authors.lastname: Tang
authors.affiliation: Key Laboratory Biotechnology Pharmaceutical Engineering, Wenzhou Medical University, Chashan University Park, Wenzhou, 325035, China.
authors.forename: Long
authors.initials: L
authors.lastname: Zheng
authors.affiliation: Key Laboratory Biotechnology Pharmaceutical Engineering, Wenzhou Medical University, Chashan University Park, Wenzhou, 325035, China.
authors.forename: Lulu
authors.initials: L
authors.lastname: Zheng
authors.affiliation: Key Laboratory Biotechnology Pharmaceutical Engineering, Wenzhou Medical University, Chashan University Park, Wenzhou, 325035, China.
authors.forename: Ting
authors.initials: T
authors.lastname: Yang
authors.affiliation: Key Laboratory Biotechnology Pharmaceutical Engineering, Wenzhou Medical University, Chashan University Park, Wenzhou, 325035, China.
authors.forename: Bingjie
authors.initials: B
authors.lastname: Yu
authors.affiliation: Key Laboratory Biotechnology Pharmaceutical Engineering, Wenzhou Medical University, Chashan University Park, Wenzhou, 325035, China.
authors.forename: Zhitao
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Key Laboratory Biotechnology Pharmaceutical Engineering, Wenzhou Medical University, Chashan University Park, Wenzhou, 325035, China.
authors.forename: Peng
authors.initials: P
authors.lastname: Lin
authors.affiliation: Key Laboratory Biotechnology Pharmaceutical Engineering, Wenzhou Medical University, Chashan University Park, Wenzhou, 325035, China.
authors.forename: Xiaokun
authors.initials: X
authors.lastname: Li
authors.affiliation: Key Laboratory Biotechnology Pharmaceutical Engineering, Wenzhou Medical University, Chashan University Park, Wenzhou, 325035, China.
authors.forename: Xiaojie
authors.initials: X
authors.lastname: Wang
mesh_terms: D000818:Animals; D017439:Cicatrix, Hypertrophic / Q000188:drug therapy*; D004195:Disease Models, Animal; D004926:Escherichia coli / Q000235:genetics*; D016222:Fibroblast Growth Factor 2 / Q000235:genetics*; D015696:Gene Products, tat / Q000235:genetics*; D051379:Mice; D008807:Mice, Inbred BALB C; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000627:therapeutic use*
chemical_list: D015696:Gene Products, tat; D011993:Recombinant Fusion Proteins; D016222:Fibroblast Growth Factor 2