title: ΦC31 integrase mediates efficient site-specific integration in sheep fibroblasts.
abstract: ΦC31 integrase, a site-specific recombinase, can effectively mediate the integration of foreign genes bearing an attB sequence into pseudo attP sites of genomes in human, mouse, and Drosophila cells. In this study, we measured ΦC31 integrase-mediated homologous recombination between attB and pseudo attP sites in sheep cells. The integration efficiency of the EGFP expression cassette with the attB sequence increased at least 2-fold in sheep fibroblasts. Three pseudo-attP sites were identified in the sheep genome, located in the intergenic regions on chromosomes 4, 13, and 7 respectively. Moreover, the transgene that was integrated at the three pseudo attP sites exhibited high levels of expression. Our study indicates that the ΦC31 integrase system provides an efficient integration tool for genetic engineering of the sheep genome.
authors.affiliation: College of Animal Science and Technology, Shihezi University, Shihezi 832003, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Ni
authors.forename: Shengwei
authors.initials: S
authors.lastname: Hu
authors.forename: Jun
authors.initials: J
authors.lastname: Qiao
authors.forename: Yuanzhi
authors.initials: Y
authors.lastname: Wang
authors.forename: Huijun
authors.initials: H
authors.lastname: Shi
authors.forename: Yong
authors.initials: Y
authors.lastname: Wang
authors.forename: Zhirui
authors.initials: Z
authors.lastname: He
authors.forename: Guozhong
authors.initials: G
authors.lastname: Li
authors.forename: Chuangfu
authors.initials: C
authors.lastname: Chen
mesh_terms: D000818:Animals; D001287:Attachment Sites, Microbiological; D005347:Fibroblasts / Q000378:metabolism*; D018014:Gene Transfer Techniques; D023281:Genomics; D049452:Green Fluorescent Proteins / Q000235:genetics; D019426:Integrases / Q000378:metabolism*; D012756:Sheep; D019076:Transgenes / Q000235:genetics
chemical_list: C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D019426:Integrases