关键词: ΦC31整合酶, 位点特异性重组, 基因操作, 卵透明带糖蛋白3, 卵母细胞

Abstract: Streptomyces phage ΦC31 integrase is a site-specific recombinase, which can catalyze site-specific, unidirectional recombination between the attP site and attB site. To explore whether it can be used to mediate the recombination of specific gene in oocytes, GV-stage oocytes were collected from 3-week-old Kunming White mice by puncturing antral follocles with a sharp needle, and micro-injected with oocyte-specific expressing ΦC31 integrase vector pZP3-INT and site -specific recombination detection vector pBCPB⁺. ΦC31 integrase mRNA were detected by RT-PCR and the recombination of pBCPB⁺ was evaluated by PCR in mouse oocytes at 48 h after injection. Both can get corresponding bands. These results indicated that the expression of ΦC31 integrase can be driven by ZP3 promoter efficiently and ΦC31 integrase can mediate the site-specific recombination between attP site and attB site in mouse GV-stage oocytes. It could be a powerful tool for the study of recombination of specific gene in mouse oocytes and would provide an alternative way for the mouse oocyte genome manipulation.