Yamao M, Katayama N, Nakazawa H, Yamakawa M, Hayashi Y, Hara S, Kamei K, Mori H
Department of Applied Biology, Kyoto Institute of Technology, Sakyo-ku, Kyoto, Japan 606-8585.
Genes Dev. 1999 Mar 1;13(5):511-6. doi: 10.1101/gad.13.5.511.
The Bombyx mori fibroin light (L)-chain gene was cloned and the green fluorescent protein (GFP) gene inserted into exon 7. The chimeric L-chain-GFP gene was used to replace the polyhedrin gene of Autographa californica nucleopolyhedrovirus (AcNPV). This recombinant virus was used to target the L-chain-GFP gene to the L-chain region of the silkworm genome. Female moths were infected with the recombinant virus and then mated with normal male moths. Genomic DNA from their progenies was screened for the desired targeting event. This analysis showed that the chimeric gene had integrated into the L-chain gene on the genome by homologous recombination and was stably transmitted through generations. The chimeric gene was expressed in the posterior silk gland, and the gene product was spun into the cocoon layer.
克隆了家蚕丝素轻（L）链基因，并将绿色荧光蛋白（GFP）基因插入到第7外显子中。嵌合的L链-GFP基因用于替换苜蓿银纹夜蛾核型多角体病毒（AcNPV）的多角体蛋白基因。这种重组病毒用于将L链-GFP基因靶向家蚕基因组的L链区域。用重组病毒感染雌蛾，然后使其与正常雄蛾交配。对其后代的基因组DNA进行筛选，以寻找所需的靶向事件。该分析表明，嵌合基因已通过同源重组整合到基因组的L链基因中，并能稳定地代代相传。嵌合基因在后丝腺中表达，其基因产物被纺入茧层。
