Disease Resistance Transferred Gene Sheep Embryo Production, Transplantation And Future Generations Biosafety Evaluation Research

This paper investigates the sheep in vivo embryo production of prokaryotic DNA microinjection and transplantation, applied research of sheep estrus synchronization, superovulation, embryo transfer of endoscopic method, and study the influence of of superovulated program, species, number of embryos transferred, season, TLR4and VP1exogenous genes on the nuclear embryo production efficiency and pregnancy rate, And genetically modified sheep, physiological and biochemical indices were tested by the data analysis of transgenic sheep blood physiological parameters and growth and development indicators. Aims to improve the DNA microinjection to produce transgenic sheep production efficiency, preliminary transgenic progeny for biological safety assessment of genetically modified sheep, and lay the foundation of speeding up the disease-resistant transgenic nurture.To further enhance the high quality meat sheep production and disease resistance of DNA microinjection transfer of TLR4of VP1gene sheep production efficiency, this test compares,the CIDR+FSH reduction+PMSG and PMSG+the same amount of CIDR+FSH group, two different superovulation programs in sheep superovulated Pronuclear Embryos efficiency, compares the different varieties Pronuclear Embryos efficiency of superovulation, the efficiency of superovulated donor for the receptor when the number of embryos on pregnancy rate, lambing rate, and the receptor of estrus synchronization treatment estrus or not, and the efficiency of season to lambing rate of prokaryotic injection, injection of different gene-positive rate of pregnancy, lambing rate and lamb. The results showed that:the CIDR+of FSH reduction+PMSG group for the body of the sheep of superovulation mating average corpus luteum number, the average received number of eggs, the average available embryo number of19.40pieces/17.80/only,17.50/only significant with higher than the CIDR+FSH same amount of+PMSG group (p<0.05);Dorset sheep superovulation effect is better than the other three varieties; superovulation better according to the weight; donor as acceptor can effectively improve the utilization of sheep and moved5～7prokaryotic embryo pregnancy rate was highest (p<0.05),45.83%, lambing rate of200.00percent, close to the efficiency of the normal receptor; in autumn sheep breeding season receptor estrus synchronization, pregnancy rate and the positive rate of43.10%,51.28%, significantly higher than the spring pregnancy rate37.77%, a positive rate of44.20%(P <0.05);nulliestrus receptor sheep pregnancy rate less than oestrus sheep (P<0.05), but for the next test for shortened sheep interval, increasing the efficiency; mixed injection of gene-positive rate is higher, but the pregnancy rate of33.33%was significantly lower than single gene is55.93%(P<0.05).This study raised production efficiency by DNA microinjection production of transgenic TLR4and VP1gene sheep, obtained a large number of transgenic offspring and positive individuals to lay the foundation for the next multiplication.In addition, we initially investigated the biosafety assessment of transgenic sheep, analyzing transgenic sheep blood physiological parameters and growth and development indicators data, Preliminarily determined the physiological effects of the transferred gene of TLR4and the anti foot and mouth disease protein gene VP1of sheep, and provide a reference for the safety of genetically modified organisms. Import TLR4gene by Southern blot analysis to determine the final6sheep were positive, the data indicate, turn TLR4gene sheep on the growth and development indicators, positive ram weight, body length and chest circumference was significantly higher (P<0.05),blood physiological parameters, especially the white blood cell count has increased, but the difference was not significant (P>0.05),this may together with improved immunity and Toll-like receptor family. Import positive sheep anti-mouth disease virus protein VP1gene in the growth and development and blood physiological indicators are negative with the transgenic negative-sheep, and differences not significant (p>0.05).Instructions to import the anti-FMD virus protein gene VP1did not impact the normal growth of the sheep.Therefore, this study improves the production of DNA microinjection transfer of TLR4of VP1gene sheep efficiency, and obtains a large number of transgenic offspring and positive individuals, and offspring blood physiological indicators and indicators of growth and development without adverse effects.