title: Effects of egt gene transfer on the development of Bombyx mori.
abstract: This study investigated the effects of gain of ecdysteroid UDP-glucosyltransferase (EGT) gene function mutation on the development of the silkworm, Bombyx mori. A novel piggyBac-derived plasmid containing the egt gene from B. mori nucleopolyhedrovirus (BmNPV) driven by a heat-shock protein (hsp) 23.7 promoter, with a neomycin-resistance gene (neo) controlled by the BmNPV ie-1 promoter and a green fluorescent protein gene (gfp) under the control of the B. mori actin 3 (A3) promoter was constructed. The vector was transferred into silkworm eggs by sperm-mediated gene transfer. Transgenic silkworms were produced after screening for neo and gfp genes and gene transfer was verified by polymerase chain reaction, dot-blot hybridization and western blotting. The hatching rate of G1 generation silkworm eggs was about 60% lower than that of normal silkworm eggs. The duration of the G1 generation larval period was extended, and the G2 generation pupal stage lasted four days longer than that in non-transgenic silkworms. The ecdysone blood level in G2 silkworms in the third instar molting stage was reduced by up to 90%. These results show that EGT suppressed transgenic silkworm molting, and that egt expression in egt-transgenic silkworms resulted in arrest of metamorphosis from pupae to moths.
authors: School of Biology & Basic Medical Science, Soochow University, Suzhou 215123, China.
authors: Xing
authors: X
authors: Zhang
authors: Renyu
authors: R
authors: Xue
authors: Guangli
authors: G
authors: Cao
authors: Xiaolong
authors: X
authors: Hu
authors: Xiaojuan
authors: X
authors: Wang
authors: Zhonghua
authors: Z
authors: Pan
authors: Min
authors: M
authors: Xie
authors: Xiaohua
authors: X
authors: Yu
authors: Chengliang
authors: C
authors: Gong
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D012831:Bombyx / Q000235:genetics* / Q000254:growth & development; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D005964:Glucosyltransferases / Q000235:genetics*; D008675:Metamorphosis, Biological / Q000235:genetics; D009154:Mutation; D017924:Nucleopolyhedroviruses / Q000235:genetics; D010957:Plasmids; D011401:Promoter Regions, Genetic
chemical_list: D005964:Glucosyltransferases; C086659:ecdysteroid UDP-glucosyltransferase