title: Germline transformation of the silkworm Bombyx mori L. by sperm-mediated gene transfer.
abstract: The domesticated silkworm Bombyx mori L. has important roles in basic biological research and applied science. To explore the practical use of transgenic technology in agricultural silkworm varieties, we fused the neomycin-resistance gene (Neo(R)) and the green fluorescent protein gene (gfp) into the piggyBac-based transposon vector and transduced it into silkworms by sperm-mediated gene transfer (SMGT). Fluorescence observation indicated the positive rate of G0 egg-batches is 72.7%. After screening against the antibiotic G418, development of individual larvae in the same brood showed significant size differences. PCR detection indicated the existence of gfp and Neo(R) and confirmed the positive rate of transgenesis as 0.47%. Southern blot analysis confirmed the presence of the exogenous genes in the genome of G7 larvae. These results show that our strategy is practical and markedly improves the efficiency of SMGT.
authors.affiliation: Sericulture Research Institute, Zhejiang Academy of Agricultural Sciences, Hangzhou, China.
authors.forename: Wenlin
authors.initials: W
authors.lastname: Zhou
authors.forename: Chengliang
authors.initials: C
authors.lastname: Gong
authors.forename: Renyu
authors.initials: R
authors.lastname: Xue
authors.forename: Guangli
authors.initials: G
authors.lastname: Cao
authors.forename: Jinru
authors.initials: J
authors.lastname: Cao
authors.forename: Aihong
authors.initials: A
authors.lastname: Ye
authors.forename: Hongbiao
authors.initials: H
authors.lastname: Weng
authors.forename: Yongqiang
authors.initials: Y
authors.lastname: Wang
mesh_terms: D000822:Animal Husbandry; D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D012831:Bombyx / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D004251:DNA Transposable Elements; D004351:Drug Resistance; D005260:Female; D018014:Gene Transfer Techniques; D017344:Genes, Insect; D005839:Gentamicins / Q000494:pharmacology; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D007814:Larva / Q000187:drug effects / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D008297:Male; D009355:Neomycin / Q000494:pharmacology; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D017924:Nucleopolyhedroviruses / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013094:Spermatozoa / Q000378:metabolism*; D014170:Transformation, Genetic
chemical_list: D000900:Anti-Bacterial Agents; D004251:DNA Transposable Elements; D005839:Gentamicins; D009419:Nerve Tissue Proteins; C569369:PGBD1 protein, human; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; C010680:antibiotic G 418; D009355:Neomycin