[Objective] This study aimed to investigate the hereditary stability of sFat-1 transgenic pigs and the differences in disease susceptivity between sFat-1 transgenic pigs and non-transgenic pigs. [Method] The integrati...[Objective] This study aimed to investigate the hereditary stability of sFat-1 transgenic pigs and the differences in disease susceptivity between sFat-1 transgenic pigs and non-transgenic pigs. [Method] The integration of sFat-1 gene in pigs was detected by PCR; the infection of transgenic pig to pseudorabies, leptospirosis, swine dysentery, brucellosis, Mycobacterium tuberculosis, rotavirus and mycoplasma hyopneumoniae was detected by using ELISA and PCR. [Result] The positive ratio of F3 generation sFat-1 transgenic pigs was 18.5%; the susceptivity of positive sFat- 1 transgenic and negative pigs to seven infectious diseases showed no significant difference. [Conclusion] Exogenous gene in sFat-1 transgenic pigs can not be stably inherited. The overall physical condition of positive transgenic and negative pigs was similar.展开更多
Omega-3(ω-3) fatty acid desaturase transgenic pigs may improve carcass fatty acid composition. The use of transgenic pigs is also an excellent large animal model for studying the role of ω-3 fatty acids in the preve...Omega-3(ω-3) fatty acid desaturase transgenic pigs may improve carcass fatty acid composition. The use of transgenic pigs is also an excellent large animal model for studying the role of ω-3 fatty acids in the prevention and treatment of coronary heart disease and cancer. Transgenic pigs carrying synthesized fatty acid desaturase-1 gene (sFat-1) from Caenorhabditis briggsae by somatic cell nuclear transfer (SCNT) were produced for the first time in China. Porcine fetal fibroblast cells were transfected with a sFat-1 expression cassette by the liposome-mediated method. Transgenic embryos were reconstructed by nuclear transfer of positive cells into enucleated in vitro matured oocytes. A total of 1889 reconstructed embryos were transferred into 10 naturally cycling gilts. Nine early pregnancies were established, 7 of which went to term. Twenty-one piglets were born. The cloning efficiency was 1.1% (born piglets/transferred embryos). The integration of the sFat-1 gene was confirmed in 15 live cloned piglets by PCR and Southern blot except for 2 piglets. Expression of the sFat-1 gene in 12 of 13 piglets was detected with RT-PCR. The data demonstrates that an efficient system for sFat-1 transgenic cloned pigs was developed, which led to the successful production of piglets expressing the sFat-1 gene.展开更多
Polyunsaturated fatty acids （PUFAs） are essential for normal growth in mammals, especially the ω-3 PUFAs, which play important roles in preventing several life-threatening diseases, such as coronary heart disease a...Polyunsaturated fatty acids （PUFAs） are essential for normal growth in mammals, especially the ω-3 PUFAs, which play important roles in preventing several life-threatening diseases, such as coronary heart disease and diabetes. In this study, we aimed to investigate whether the sFat-1 gene from Caenorhabditis briggsae could be functionally expressed in transgenic pigs, and whether the transgenic could synthesize high quality ω-3 PUFAs endogenously. In this study, a gene construct consisting of CMV promoter and 1.9 kb cDNA of ω-3 fatty acid desaturase gene （sFat-1） from C. briggsae was injected into the male pronucleus of pig embryos by microinjection. The piglets were screened for the transgene by PCR, Southern blot and reverse transcription-PCR analysis. Pigs that give positive results were mated with wild-type pigs to produce the next generation and the transmission of transgene was examined by PCR analysis. Fatty acids compositions of various tissues in the transgenic pigs were then analyzed by gas chromatograph. A total of 878 embryos were transferred into 42 recipients, among which 29 successfully got pregnant and gave birth to a total of 162 piglets, and 8 of them were identified to be transgenic. Fatty acid compositions in the transgenic pigs were altered, and the levels of ω-6：ω-3 ratios were decreased from 14.53 in the control to 2.62 in Fat-1 transgenic pigs. A number of primary sFat-1-transgenic pigs were bred in this study, which lays the foundation for cultivation of new varieties of transgenic pigs.展开更多
基金Supported by National Major Program of Genetically Modified Organism for New Species Cultivation of China(2011ZX08011-004)Project from Hubei Agricultural Science and Technology Innovation Center(2011-620-001-003)~~
文摘[Objective] This study aimed to investigate the hereditary stability of sFat-1 transgenic pigs and the differences in disease susceptivity between sFat-1 transgenic pigs and non-transgenic pigs. [Method] The integration of sFat-1 gene in pigs was detected by PCR; the infection of transgenic pig to pseudorabies, leptospirosis, swine dysentery, brucellosis, Mycobacterium tuberculosis, rotavirus and mycoplasma hyopneumoniae was detected by using ELISA and PCR. [Result] The positive ratio of F3 generation sFat-1 transgenic pigs was 18.5%; the susceptivity of positive sFat- 1 transgenic and negative pigs to seven infectious diseases showed no significant difference. [Conclusion] Exogenous gene in sFat-1 transgenic pigs can not be stably inherited. The overall physical condition of positive transgenic and negative pigs was similar.
基金suppored by the Na-tional High-Tech Research and Development program (Grant No. 2006AA02Z113)
文摘Omega-3(ω-3) fatty acid desaturase transgenic pigs may improve carcass fatty acid composition. The use of transgenic pigs is also an excellent large animal model for studying the role of ω-3 fatty acids in the prevention and treatment of coronary heart disease and cancer. Transgenic pigs carrying synthesized fatty acid desaturase-1 gene (sFat-1) from Caenorhabditis briggsae by somatic cell nuclear transfer (SCNT) were produced for the first time in China. Porcine fetal fibroblast cells were transfected with a sFat-1 expression cassette by the liposome-mediated method. Transgenic embryos were reconstructed by nuclear transfer of positive cells into enucleated in vitro matured oocytes. A total of 1889 reconstructed embryos were transferred into 10 naturally cycling gilts. Nine early pregnancies were established, 7 of which went to term. Twenty-one piglets were born. The cloning efficiency was 1.1% (born piglets/transferred embryos). The integration of the sFat-1 gene was confirmed in 15 live cloned piglets by PCR and Southern blot except for 2 piglets. Expression of the sFat-1 gene in 12 of 13 piglets was detected with RT-PCR. The data demonstrates that an efficient system for sFat-1 transgenic cloned pigs was developed, which led to the successful production of piglets expressing the sFat-1 gene.
基金supported by the National Transgenic Breeding Project, China (2008ZX08010-004)the National Natural Science Foundation of China (30830080)+1 种基金the National Basic Research Program of China (G2006CB102105, 2009CB941604)the National High Technology Research and Development Program of China (20060110Z1039, 2008AA10Z143)
文摘Polyunsaturated fatty acids （PUFAs） are essential for normal growth in mammals, especially the ω-3 PUFAs, which play important roles in preventing several life-threatening diseases, such as coronary heart disease and diabetes. In this study, we aimed to investigate whether the sFat-1 gene from Caenorhabditis briggsae could be functionally expressed in transgenic pigs, and whether the transgenic could synthesize high quality ω-3 PUFAs endogenously. In this study, a gene construct consisting of CMV promoter and 1.9 kb cDNA of ω-3 fatty acid desaturase gene （sFat-1） from C. briggsae was injected into the male pronucleus of pig embryos by microinjection. The piglets were screened for the transgene by PCR, Southern blot and reverse transcription-PCR analysis. Pigs that give positive results were mated with wild-type pigs to produce the next generation and the transmission of transgene was examined by PCR analysis. Fatty acids compositions of various tissues in the transgenic pigs were then analyzed by gas chromatograph. A total of 878 embryos were transferred into 42 recipients, among which 29 successfully got pregnant and gave birth to a total of 162 piglets, and 8 of them were identified to be transgenic. Fatty acid compositions in the transgenic pigs were altered, and the levels of ω-6：ω-3 ratios were decreased from 14.53 in the control to 2.62 in Fat-1 transgenic pigs. A number of primary sFat-1-transgenic pigs were bred in this study, which lays the foundation for cultivation of new varieties of transgenic pigs.
