Li Zhen, Jiang Yue, Cao Guangli, Li Jingzhi, Xue Renyu, Gong Chengliang
School of Biology & Basic Medical Science, Soochow University, Suzhou, 215123, People's Republic of China.
Mol Biol Rep. 2015 Jan;42(1):19-25. doi: 10.1007/s11033-014-3735-z. Epub 2014 Sep 16.
A targeting vector consisting of a fusion gene of the green fluorescent protein (GFP) gene gfp and the antimicrobial peptide cecropin gene cec flanked by pieces of the 5' and 3' sequences of the fibroin L chain gene fib-L of the silkworm (Bombyx mori) and a negative selection DsRed marker gene driven by the baculovirus immediate early gene 1 (i.e.-1) promoter, was used to target the silkworm genome in order to explore the possibility of improving the performance of silk. A transgenic silkworm with a green fluorescent cocoon was obtained and PCR analysis of its genome confirmed that the target genes had been integrated into the silkworm genome correctly. Furthermore, in the posterior silk glands of the G6 generation transformation silkworm, a band representing the fusion protein Fib-L-GFP-Cec with a molecular mass of 68.7 kDa was detected by western blotting with an antibody against GFP. An investigation of the number of bacteria attached to a cocoon showed the transgenic silkworm cocoon possessed antibacterial properties. These results suggested the performance of silk can be improved by modifying the fibroin gene.
一种靶向载体,由绿色荧光蛋白(GFP)基因gfp和抗菌肽天蚕素基因cec的融合基因组成,两侧是家蚕(Bombyx mori)丝素轻链基因fib-L的5'和3'序列片段,以及由杆状病毒立即早期基因1(ie-1)启动子驱动的负选择DsRed标记基因,用于靶向家蚕基因组,以探索改善蚕丝性能的可能性。获得了具有绿色荧光茧的转基因家蚕,其基因组的PCR分析证实目标基因已正确整合到家蚕基因组中。此外,在G6代转化家蚕的后部丝腺中,用抗GFP抗体进行蛋白质免疫印迹检测到一条分子量为68.7 kDa的代表融合蛋白Fib-L-GFP-Cec的条带。对附着在茧上的细菌数量的调查表明,转基因家蚕茧具有抗菌特性。这些结果表明,通过修饰丝素基因可以改善蚕丝的性能。