关键词: 家蚕, piggyBac, 转基因技术, 孵化率, 转化率, 蚕品种, 供体质粒, 辅助质粒, 显微注射, 荧光筛选, 荧光标记

Abstract: Efficient transgenic technology of silkworm is the key to promote the functional research of silkworm genome, and to solve the major problems in the sericulture industry. On the basis of the established transgenic technology in silkworm, we investigated the effect of several factors on transformation efficiency. The results showed that when taking P50 as a recipient strain, pBac［GOI］as a donor plasmid and pHA4PIG as a helper plasmid, and red fluorescent protein gene, DsRed, driven by an eye-specific promoter, 3×p3, as a reporter gene, microinjection into the eggs during 2-3 h after oviposition increased the transformation efficiency. The hatching rate of injected eggs and the transformation rate were 62.7% and 34.8%, respectively. The best time for fluorescent screening was the 5th to 8th day of embryo. The results also showed that when varied from 2 000 to 8 000 bp the length of inserts had no significant effect on transformation rate. The results will set a model for transgensis of other lepidopteran insects and will contribute to the functional genomics study of the silkworm and the development of molecular bioreactors involved.

Key words: Bombyx mori, piggyBac, transgenic technology, hatching rate, transformation rate, silkworm strains, donor plasmid, helper plasmid, microinjection, fluorescent screening, fluorescent marker