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Abstract As a member of transforming growth factor(TGF-β), myostatin(MSTN) plays an important role for the specific regulation of muscle growth of animals. The purpose of this study is to verify the efficiency of zinc-finger nucleases(ZFN) gene editing on porcine(Sus scrofa) and to pick out the efficient gene whicn includes targeting sites. Using the method of microinjection, ZFN plasmid were injected that were specifically targeted on MSTN into porcine oocyte parthenogenetic embryo . Then the MSTN sequence of cultured embryos were amplified and analyzed by PCR. The results showed that totally 373 embryos were injected, and 2 mutate sequences were found after the specifically amplification by MSTN primers which disigned according JN630464(GenBank accession number), 14 single base mutations were found in No. 39 embryo, and 11 single base mutations were found in No. 321 embryo. Both mutations occurred in ZFN targeting sites, the targeting efficiency was about 0.54%. The results ensure the possibility and provide feasibility analysis for the preparation of MSTN knockout pig by ZFN gene editing technology.
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Received: 09 December 2013
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