title: Large-scale production of bioactive recombinant human acidic fibroblast growth factor in transgenic silkworm cocoons.
abstract: With an increasing clinical demand for functional therapeutic proteins every year, there is an increasing requirement for the massive production of bioactive recombinant human acidic fibroblast growth factor (r-haFGF). In this present study, we delicately explore a strategy for the mass production of r-haFGF protein with biological activity in the transgenic silkworm cocoons. The sequence-optimized haFGF was inserted into an enhanced sericin-1 expression system to generate the original transgenic silkworm strain, which was then further crossed with a PIG jumpstarter strain to achieve the remobilization of the expression cassette to a "safe harbor" locus in the genome for the efficient expression of r-haFGF. In consequence, the expression of r-haFGF protein in the mutant line achieved a 5.6-fold increase compared to the original strain. The high content of r-haFGF facilitated its purification and large-scald yields. Furthermore, the r-haFGF protein bioactively promoted the growth, proliferation and migration of NIH/3T3 cells, suggesting the r-haFGF protein possessed native mitogenic activity and the potential for wound healing. These results show that the silk gland of silkworm could be an efficient bioreactor strategy for recombinant production of bioactive haFGF in silkworm cocoons.
authors.affiliation: State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing, China.|College of biotechnology, Southwest University, Chongqing, China.
authors.forename: Feng
authors.initials: F
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing, China.
authors.forename: Riyuan
authors.initials: R
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing, China.|College of biotechnology, Southwest University, Chongqing, China.
authors.forename: Yuancheng
authors.initials: Y
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing, China.|College of biotechnology, Southwest University, Chongqing, China.
authors.forename: Ping
authors.initials: P
authors.lastname: Zhao
authors.affiliation: State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing, China.|College of biotechnology, Southwest University, Chongqing, China.
authors.forename: Qingyou
authors.initials: Q
authors.lastname: Xia
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D019149:Bioreactors; D012831:Bombyx / Q000235:genetics*; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D004251:DNA Transposable Elements; D016220:Fibroblast Growth Factor 1 / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology; D015870:Gene Expression; D006801:Humans; D051379:Mice; D041681:NIH 3T3 Cells; D058849:Protein Refolding; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology
chemical_list: D004251:DNA Transposable Elements; D011994:Recombinant Proteins; D016220:Fibroblast Growth Factor 1