title: The Escherichia coli-derived thymosin β4 concatemer promotes cell proliferation and healing wound in mice.
abstract: Thymosin β4 (Tβ4) is one of the most promising thymosins for future clinical applications, and it is anticipated that commercial demand for Tβ4 will increase. In order to develop a new approach to produce recombinant Tβ4, a 168 bp DNA (termed Tβ4) was designed based on the Tβ4 protein sequence and used to express a 4 × Tβ 4 concatemer (four tandem copies of Tβ4, termed 4 × Tβ4) together with a histidine tag (6 × His) in E. coli (strain BL21). SDS-PAGE and western blot analysis were used to confirm that a recombinant 4 × Tβ4 protein of the expected size (30.87 kDa) was produced following the induction of the bacterial cultures with isopropyl β-D-thiogalactoside (IPTG). The E. coli-derived 4 × Tβ4 was purified by Ni-NTA resin, and its activities were examined with regard to both stimulating proliferation of the mice spleen cells in vitro and in vivo wound healing. The results demonstrate that these activities of the E. coli-derived recombinant 4 × Tβ4 were similar or even better than existing commercially obtained Tβ4. This production strategy therefore represents a potentially valuable approach for future commercial production of recombinant Tβ4.
authors.affiliation: Plant Biotechnology Research Center, School of Agriculture and Biology, Fudan-SJTU-Nottingham, Plant Biotechnology R and D Center, Shanghai Jiao Tong University, Shanghai 200240, China.
authors.forename: Xiaolei
authors.initials: X
authors.lastname: Wang
authors.forename: Guihua
authors.initials: G
authors.lastname: Yang
authors.forename: Shanshuang
authors.initials: S
authors.lastname: Li
authors.forename: Meifeng
authors.initials: M
authors.lastname: Gao
authors.forename: Pangfeng
authors.initials: P
authors.lastname: Zhao
authors.forename: Lingxia
authors.initials: L
authors.lastname: Zhao
mesh_terms: D000818:Animals; D001483:Base Sequence; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D044708:DNA, Concatenated / Q000494:pharmacology*; D004848:Epithelium / Q000098:blood supply / Q000187:drug effects / Q000473:pathology; D004926:Escherichia coli / Q000378:metabolism*; D015603:Keratinocytes / Q000187:drug effects / Q000473:pathology; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology; D013947:Thymosin / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology*; D014945:Wound Healing / Q000187:drug effects*; D000099201:Thymosin beta(4)
chemical_list: D044708:DNA, Concatenated; D011994:Recombinant Proteins; D013947:Thymosin; D000099201:Thymosin beta(4)