Production of Phytase Transgenic Porcine Blastocysts by Handmade Cloning

To address the problem of the phosphorus pollution from livestocks, phytase transgenic animals have been developed by somatic cell nuclear transfer (SCNT). However, the SCNT technology is quite complicated, inefficient and requires expensive micromanipulators. In this study, we used the economical handmade cloning (HMC) technique to produce phytase transgenic porcine blastocysts. The recombinant eukaryotic expression plasmid containing the modified phytase gene was generated and transferred into the donor porcine fetal fibroblasts. The reconstructed embryos were produced by the use of transgenic donor cells and cytoplasms from porcine mature oocytes. The transgenic blastocysts were obtained after subsequent culture of the reconstructed embryos. The reverse transcription-polymerase chain reaction (RT-PCR) identification validated the successful expression of phytase gene in the transgenic blastocysts at the transcriptional level. These handmade cloned blastocysts provide the fundamentals for further producing phytase transgenic pigs with potential to be environmentally friendly.