Study on the Production of Porcine Phytase Transgenic Embryos with Somatic Cell Nuclear Transfer |
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| Authors: | DAI Jian-jun WU Cai-feng ZHANG Shu-shan GU Xiao-long ZHANG Ting-yu WU Zhi-qiang ZHANG De-fu |
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| Institution: | 1. Institute of Animal Science and Veterinary Medicine, Shanghai Academy of Agricultural Sciences, Shanghai 201106, China;2. Division of Animal Genetic Engineering, Shanghai Municipal Key Laboratory of Agri-genetics and Breeding, Shanghai 201106, China |
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| Abstract: | In order to get porcine phytase salivary glands specific express cloned embryos,this study used phytase specific expressed plasmid DNA (including the promotor and terminater sequences of parotid secretary protein (PSP) gene,Neo selected gene and EGFP report gene and high enzymatic activity phytase gene (appA)) and transfected it into porcine fetal fibroblast cells with liposome.The stably transfected cell lines were screened by G418 selection,and positive phytase transgenic embryos were constructured by somatic cell nuclear transfer.The results showed that the phytase plasmid of this experiment could be used to carry on the cell selection,and the more smaller of plasmid DNA size,the more higher transfected efficiency was got.The plasmid YM6552 with 14.89 kb size only got 7.1% transfected efficiency,while EGPF plasmid got 43.4%.In the formation of single cell colony,pYN3600 (more smaller DNA size) got 25 colony numbers,and 14 of them expressed green fluorescent protein,11 of them were phytase PCR positive.They were much higher than those of YM6552 (19,8 and 6).All the reconstructed transgenic embryoes from the positive cell lines were expressed green fluorescent protein.Though the in vitro development ability of transgenic cloned embryos were a little lower than that of control group,there were no significant differences between them (P>0.05).In conclusion,all the phytase plasmid,cell selection method and nuclear transfer techniques could be used to produce porcine phytase salivary glands specific express embryos. |
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| Keywords: | phytase parotid secretary protein transgenic nuclear transfer porcine |
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