PiggyBac transposon has demonstrated its long-term and stable transposition on genomes of various species but lacking of the evidence on farm animal genomes. In this study, we constructed a piggyBac transposon marked ...PiggyBac transposon has demonstrated its long-term and stable transposition on genomes of various species but lacking of the evidence on farm animal genomes. In this study, we constructed a piggyBac transposon marked with enhanced green fluorescent protein （eGFP） and showed efficient transposition in porcine somatic cells and cloned embryos. Our results demonstrated that piggyBac transposase could efficiently catalyze transposition in porcine fetal fibroblast cells, as well as in embryos. PiggyBac transposition generated 18-fold more eGFP-positive cell colonies compared to pEGFP-C1 random insertion mutagenesis, but excessive transposase might affect the transfection rate. Also piggyBac mediated 4-fold more eGFP expression than random insertion in cells and 17-fold in cloned embryos at mRNA level. When the mutagenized cells were used for somatic cell nuclear transfer （SCNT）, the cleavage rate and blastocyst rate of constructed embryos harboring piggyBac transposition had no difference with random insertion group. This study provides key information on the piggyBac transposon system as a tool for creating transgenic pigs.展开更多
Tissue culture seedlings of the hybrid Cymbidium were inoculated with six different fungal strains, isolated from the roots of different wild terrestrial orchids. About three months later, the average increment of fre...Tissue culture seedlings of the hybrid Cymbidium were inoculated with six different fungal strains, isolated from the roots of different wild terrestrial orchids. About three months later, the average increment of fresh weight of seedlings inoculated with strains CF1, CF3 and CF12 were respectively 130.26%, 345.65% and 153.34% while that of the control was only 88.40%. The differences between the three treatments and the control were statistically significant （α = 0.05）, highlighting the treatment with strain CF3 （α = 0.01）. In addition, the three strains were obtained by re-isolating. Pelotons, regarded as typical structures of orchid mycorrhiza, were also found in the inoculating roots under a microscope. It seems that the strains of CF1, CF3, and CF12 are associated with the hybrid Cymbidium and supplied the orchid with nutrition. It can be confirmed that the three strains are beneficial for the seedlings of this hybrid.展开更多
基金Supported by the National Projects of Genetic Modified Organism Breeding Technology (2008ZX08006-002)the State Transgenic Research Programme of China (2008ZX08006-002)
文摘PiggyBac transposon has demonstrated its long-term and stable transposition on genomes of various species but lacking of the evidence on farm animal genomes. In this study, we constructed a piggyBac transposon marked with enhanced green fluorescent protein （eGFP） and showed efficient transposition in porcine somatic cells and cloned embryos. Our results demonstrated that piggyBac transposase could efficiently catalyze transposition in porcine fetal fibroblast cells, as well as in embryos. PiggyBac transposition generated 18-fold more eGFP-positive cell colonies compared to pEGFP-C1 random insertion mutagenesis, but excessive transposase might affect the transfection rate. Also piggyBac mediated 4-fold more eGFP expression than random insertion in cells and 17-fold in cloned embryos at mRNA level. When the mutagenized cells were used for somatic cell nuclear transfer （SCNT）, the cleavage rate and blastocyst rate of constructed embryos harboring piggyBac transposition had no difference with random insertion group. This study provides key information on the piggyBac transposon system as a tool for creating transgenic pigs.
文摘Tissue culture seedlings of the hybrid Cymbidium were inoculated with six different fungal strains, isolated from the roots of different wild terrestrial orchids. About three months later, the average increment of fresh weight of seedlings inoculated with strains CF1, CF3 and CF12 were respectively 130.26%, 345.65% and 153.34% while that of the control was only 88.40%. The differences between the three treatments and the control were statistically significant （α = 0.05）, highlighting the treatment with strain CF3 （α = 0.01）. In addition, the three strains were obtained by re-isolating. Pelotons, regarded as typical structures of orchid mycorrhiza, were also found in the inoculating roots under a microscope. It seems that the strains of CF1, CF3, and CF12 are associated with the hybrid Cymbidium and supplied the orchid with nutrition. It can be confirmed that the three strains are beneficial for the seedlings of this hybrid.
