Transgenic animal mammary gland bioreactors are used to produce recombinant proteins with appropriate post-translational modifications.The nuclear transfer of transgenic somatic cells is a powerful method to produce m...Transgenic animal mammary gland bioreactors are used to produce recombinant proteins with appropriate post-translational modifications.The nuclear transfer of transgenic somatic cells is a powerful method to produce mammary gland bioreactors.We established an efficient gene transfer and nuclear transfer approach in goat somatic cells.Gene targeting vector pGBC2LF was constructed by cloning human lactoferrin(LF)gene cDNA into exon 2 of the milk goat beta-casein gene and the endogenous start codon was replaced by that of human LF gene.Goat fetal fibroblasts were transfected with lin-earized pGBC2LF and 14 cell lines were positive accord-ing to PCR and Southern blot.The transgenic cells were used as donor cells of nuclear transfer and some of recon-structed embryos could develop into blastocyst in vitro.展开更多
基金This work was supported by grants from the National High Technology Research and Development Program of China(No.2002AA206621)the Key Programs for Science and Technology Development of Shandong Province(No.J05K06)+2 种基金the Program of Science and Technology Grant of Qingdao City(No.06-2-2-11-jch)Nature Science Foundation of Shandong Province(No.Z2007D01)Doctoral Foundation of Qingdao Agricultural University(No.630615).
文摘Transgenic animal mammary gland bioreactors are used to produce recombinant proteins with appropriate post-translational modifications.The nuclear transfer of transgenic somatic cells is a powerful method to produce mammary gland bioreactors.We established an efficient gene transfer and nuclear transfer approach in goat somatic cells.Gene targeting vector pGBC2LF was constructed by cloning human lactoferrin(LF)gene cDNA into exon 2 of the milk goat beta-casein gene and the endogenous start codon was replaced by that of human LF gene.Goat fetal fibroblasts were transfected with lin-earized pGBC2LF and 14 cell lines were positive accord-ing to PCR and Southern blot.The transgenic cells were used as donor cells of nuclear transfer and some of recon-structed embryos could develop into blastocyst in vitro.