Influence of oct4 on Early Embryonic Development of Mouse-Porcine Interspecies Nuclear Transfer
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R321

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    Abstract:

    Objective To explore whether the Oct4 transcription factor can improve the mouse-porcine interspecies nuclear transfer development or not.Methods Oct4 cDNA was amplified by RT-PCR.Two recombinant expression plasmids were constructed as the pEGFP-N1-Oct4-EGFP fusion vector and pEGFP-N-1-Oct4-EGFP termination vector,and pEGFP-N1 as the negative control,then transfect the NIH3T3 cell with lipofectamineTM2000.RT-PCR assay showed that oct4 DNA fragment could be amplified,and confirmed by fluorescence microscopy.Then the cell was inserted into the perivitelline space of the enucleated porcine oocyte to investigate the development of interspecies embryo.Results All the non-transgenic NIH3T3 group,transgenic Oct4-EGFP NIH3T3 and pEGFP-N-1 groups could support the early development of cloned interspecies embryo,and the cleavage rate and development rate into the eight-cell stage were not significantly different.Conclusion It is clear that NIH3T3 can support the early embryonic development of mouse-porcine interspecies nuclear transfer.But maybe due to the low development of NIH3T3 derived nuclear transfer embryo and our limited condition of micromanipulation,the Oct4 gene does not have an improving effect on interspecies nuclear transfer.Further studies are still needed to elucidate its concrete mechanism.

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  • Received:
  • Revised:May 09,2006
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