利用转基因家蚕在蚕丝中表达重组人表皮生长因子

Expression of Recombinant Human Epidermal Growth Factor in Silk by Transgenic Silkworm

人表皮生长因子(human epidermal growth factor,hEGF)具有刺激细胞增殖的强大能力,是伤口愈合的关键因子。利用转基因技术,在家蚕后部丝腺利用丝素轻链基因(fibroin light chain,FibL)启动子驱动FibL-hEGF融合蛋白的表达。借助piggyBac转座子将FibL-hEGF融合基因导入到家蚕基因组,利用荧光检测总共获得8个转基因阳性卵圈,转基因阳性率为36.36%。PCR和反向PCR检测证明外源融合基因FibL-hEGF已被成功整合到家蚕基因组中,且随机插入到家蚕染色体的5个位点。Western blotting检测表明FibL-hEGF融合蛋白在家蚕后部丝腺中表达且成功分泌到蚕茧中,进一步证实了转基因家蚕的产生。本研究中利用piggyBac转座子介导的转基因技术成功获得了转基因家蚕并得到重组hEGF蚕丝,该蚕丝可用于制造伤口敷料。

Human epidermal growth factor(hEGF)is a key factor involved in wound healing owing to its powerful ability to stimulate cell proliferation.Transgenic technology was used to express FibL-hEGF fusion protein driven by the fibroin light chain gene(FibL)promoter in the posterior silk glands.FibL-hEGF fusion gene was introduced into silkworm genome by piggyBac transposon.A total of eight transgenic positive broods were obtained by fluorescence detection,with the transgenic positive rate as 36.36%.PCR and inverse PCR results showed that the exogenous fusion gene FibL-hEGF had been successfully integrated into the silkworm genome with five random insertion sites on the silkworm chromosomes.Western blotting detection showed that FibL-hEGF fusion protein was expressed in the posterior silk gland of silkworm and successfully secreted into cocoon,further confirming the generation of transgenic silkworms.Therefore,transgenic silkworms and recombinant hEGF silk were successfully obtained by piggyBac transposon-mediated transgenic technology in this study,which would have the potential for wound dressings fabrication.