title: FLP recombinase-mediated site-specific recombination in silkworm, Bombyx mori.
abstract: A comprehensive understanding of gene function and the production of site-specific genetically modified mutants are two major goals of genetic engineering in the post-genomic era. Although site-specific recombination systems have been powerful tools for genome manipulation of many organisms, they have not yet been established for use in the manipulation of the silkworm Bombyx mori genome. In this study, we achieved site-specific excision of a target gene at predefined chromosomal sites in the silkworm using a FLP/FRT site-specific recombination system. We first constructed two stable transgenic target silkworm strains that both contain a single copy of the transgene construct comprising a target gene expression cassette flanked by FRT sites. Using pre-blastoderm microinjection of a FLP recombinase helper expression vector, 32 G3 site-specific recombinant transgenic individuals were isolated from five of 143 broods. The average frequency of FLP recombinase-mediated site-specific excision in the two target strains genome was approximately 3.5%. This study shows that it is feasible to achieve site-specific recombination in silkworms using the FLP/FRT system. We conclude that the FLP/FRT system is a useful tool for genome manipulation in the silkworm. Furthermore, this is the first reported use of the FLP/FRT system for the genetic manipulation of a lepidopteran genome and thus provides a useful reference for the establishment of genome manipulation technologies in other lepidopteran species.
authors: State Key Laboratory of Silkworm Genome Biology, Institute of Sericulture and Systems Biology, Southwest University, Chongqing, China.
authors: Ding-Pei
authors: DP
authors: Long
authors: Ai-Chun
authors: AC
authors: Zhao
authors: Xue-Jiao
authors: XJ
authors: Chen
authors: Yang
authors: Y
authors: Zhang
authors: Wei-Jian
authors: WJ
authors: Lu
authors: Qing
authors: Q
authors: Guo
authors: Alfred M
authors: AM
authors: Handler
authors: Zhong-Huai
authors: ZH
authors: Xiang
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D001483:Base Sequence; D012831:Bombyx / Q000196:embryology / Q000235:genetics*; D004254:DNA Nucleotidyltransferases / Q000378:metabolism*; D004625:Embryo, Nonmammalian / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors / Q000008:administration & dosage / Q000235:genetics; D049750:Genome, Insect / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics; D007267:Injections; D008164:Luminescent Proteins / Q000235:genetics; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional / Q000235:genetics; D011995:Recombination, Genetic; D015203:Reproducibility of Results
chemical_list: D008164:Luminescent Proteins; C485184:enhanced green fluorescent protein; C413663:fluorescent protein 583; D049452:Green Fluorescent Proteins; D004254:DNA Nucleotidyltransferases; C045119:FLP recombinase