【作者】 成荣；

【导师】 陈国宏； 李建凡；

【作者基本信息】 扬州大学 ， 动物遗传育种与繁殖， 2004， 硕士

【摘要】 本研究以LacZ基因为报告基因，构建了两种载体质粒：p-CMV-β-Gal和p-OV-β-Gal，分别利用显微注射法和精子介导法将这两种载体质粒转移到鸡早期胚胎内，对外源基因的整合及表达情况进行检测分析。同时，对添加抗菌素溶液是否有利于鸡胚的正常发育进行了探讨。结果如下： 1、在处理过的受精蛋中添加一定量的抗菌素(青、链霉素)溶液对于提高鸡胚抵御外部环境污染能力、避免鸡胚早期死亡的效果并不显著，但在实际试验过程中发现添加的抗菌素溶液能起到补充水分的作用，保证鸡胚的正常发育。 2、利用显微注射法转移载体质粒(p-CMV-β-Gal)的过程破坏了受精蛋的结构，对鸡胚的发育造成影响，表现在鸡胚在早期孵化阶段大批死亡，死亡率高达58.07％(367／632)，但这种方法能够将载体质粒转移到鸡早期胚胎内，外源基因整合阳性率为21.43％(15／70)，在部分鸡胚中能检测到外源基因的表达，阳性率为9.52％(4／42)。 3、利用精子介导法能将载体质粒(p-OV-β-Gal)转移到鸡早期胚胎内，外源基因整合阳性率为5.91％(11／186)，表达阳性率为16.94％(41／242)。 以上试验结果显示：显微注射法和精子介导法均能将载体质粒转移到鸡早期胚胎中，外源基因能被整合到宿主基因组中，有些能进一步得到表达。说明这两种方法均可作为制备转基因鸡的研究方法。更多还原

【Abstract】 To explore the possibility of generation of transgenic chicken, two vectors (p-CMV- β -Gal and p-OV- 3 -Gal) of LacZ gene as report gene were transmitted into the chicken’s early embryo by two methods: Microinjection Method and Sperm-mediated Method, then the LacZ gene’s integration and expression were monitored. At the same time, the effect of bacteriophage was discussed. The results were as showed:1. After working the egg, a litter of bacteriophage liquor was appended into the egg. Its effect of enhancing the immunity of the embryo and avoiding the embryo dying early was not remarkable. But during the course of the experiment, another effect was discovered: The appended liquor could supply the wet that quickly volatilized in the course of hatching.2. During the course of microinjecting, the structure of the egg was broken. Itresulted in that the growth of the embryo was affected and many embryos died early, the death percentage was 58.07%(367/632). In fact, the vector (p-CMV-β-Gal) could be transmitted into the chicken’s early embryo by Microinjection Method, and the percentage of LacZ gene’s integration and expression respectively were 21.43%(15/7o)and 9.52 %(4/42)-3. The vector (p-OV- β -Gal) could be transmitted into the chicken’s early embryo by sperm-mediated method, the percentage of LacZ gene’s integration and expression respectively were 5.91 %(n/,86) and 16.94%(41/242).From the results of the experiments, it could be concluded that two vectors could be transmitted into the chicken’s early embryo by microinjection method or sperm-mediated method, and the LacZ gene could be integrated and expressed. It was showed that the two methods could be used for generation of the transgenic chicken.更多还原