Expression Of Human Basic Fibroblast Growth Factor In Silk Glands Of Silkworm

The silkworm gland bioreactor has been used generally to express exogenous protein by silk glands which generate high-yielding as well as good quality synthetic secretion silk protein.Human fibroblast growth factor(FGFs)play a crucial regulatory role in human cell tissue regeneration and wound healing.This study aim to targeting human basic fibroblast growth factor(hbFGF)into silkworm genome by the piggyBac transposon,finally we acquire the composite silk which are combined with hbFGF depending on the genetically modified technology.We construct a piggyBac transposon with two expression boxes by one plasmid,one expresses hbFGF by heavy chain promotor,another one expresses a enhanced green fluorescent protein gene(EGFP)by IE1 promotor.With the help of EGFP,genetically modified positive silkworms could be selected by the fluorescent electron microscope.pB-hbFGF and helper plasmid are mixed to micro-injected into the silkworm eggs,named as Generation 0(GO).Positive individuals could be detected at once by EGFP when Generation 1(G1)eggs hatch as silkworm.Positive individuals are feeded untill they secrete hbFGF silk.Study shows:Firstly,PCR confirmes that hbFGF has been integrated into the silkworm genome.Then fluorescent quantitative experiments suggest hbFGF silkworm express hbFGF mRNA far more than non-transgmogenic Lan10.And then hbFGF silk is detected 34kD hbFGF protein by western blot.Finally silk mechanical properties statistic show that hbFGF silk harder more than non-transgenic silk Lan10.In short,we creates a stable genetic linage,hbFGF silkworm which secretes hbFGF silk.Universally acknowledged that hbFGF accelerate cell tissue regeneration and wound healing,most impotantly,hbFGF silk has improved mechanical properties than non-transgenic,all of the research develope the posibility of innovative surgical sutures that combines mutiple hbFGF function with improved mechanical properties of silk without removal from patients.