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

doi:10.3724/SP.J.1005.2009.00595

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Hereditas(Beijing) >

2009 , Vol. 31 >Issue 6: 595 - 599

DOI: https://doi.org/10.3724/SP.J.1005.2009.00595

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

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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 date: 2009-03-19

Revised date: 2009-04-10

Online published: 2009-06-10

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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.

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XU Huan-Yu, GONG Xiu-Li, GUO Xin-Bing, MA Qing-Wen, ZENG Yi-Tao . Construction of the oocyte-specific expressing fC31 integrase vector pZP3-INT and its expression in mouse oocytes[J]. Hereditas(Beijing), 2009 , 31(6) : 595 -599 . DOI: 10.3724/SP.J.1005.2009.00595

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