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HEREDITAS ›› 2009, Vol. 31 ›› Issue (6): 595-599.doi: 10.3724/SP.J.1005.2009.00595

• 研究报告 • Previous Articles     Next Articles

Construction of the oocyte-specific expressing fC31 integrase vector pZP3-INT and its expression in mouse oocytes

XU Huan-Yu;GONG Xiu-Li;GUO Xin-Bing;MA Qing-Wen;ZENG Yi-Tao   

  1. Shanghai Institute of Medical Genetics, Shanghai JiaoTong University, Key Laboratory of Embryo Molecular Biology, Ministry of Health, Shanghai Laboratory of Embryo and Reproduction Engineering, Shanghai 200040, China
  • Received:2009-03-19 Revised:2009-04-10 Online:2009-06-10 Published:2009-06-10
  • Contact: ZENG Yi-Tao

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.