摘要
采用精子载体介导法先后对 1 952只绵羊进行转基因试验 ,经PCR和South ern检测 1 592只后代中 ,转基因羊 1 0 3只 ,最高转基因效率达 9.86 % ;其中 2只转基因母羊乳汁中人胰岛素原基因表达量分别达 1 99.97mg/L和 1 83.6 2mg/L。荧光原位PCR结果证实牛、山羊和绵羊的精子经处理能携带外源基因 ,精子携带外源DNA的比率分别为 :绵羊 2 7.1 2 %、山羊 30 .57%、牛 31 .1 6 % ;同时外源基因进入到精子细胞并定位于核及其外周区域。研究还提示使用精子载体介导法时必须充分洗涤精液以除去精清 。
Within five years, transgenic sheep were produced by sperm-mediated exogenous DNA transfer method combined with Artification Insemination (AI) technique. The results of PCR and Southern blot showed that one hundred and three of 1592 offspring carried exogenous DNA. The transgenic efficiency was 9.86% in 1995. With analysis by radioimmunoassay (RIA) determined the expression of human proinsulin in the milk of two transgenic sheep were 199.97mg/L and 183.62mg/L, respectively. Fluosence in situ PCR showed that the sperm of bull, goat and sheep can carry exogenous DNA after patient treatment method and exogenous DNA enter sperm nucleus and locate in the internal and peripheral nucleus of sperm. The average transfer efficiency of PRINS for sperm was 27.12% in sheep, 31.16% in bull, 30.57% in goat, respectably. We also found that the sperm must be washed completely to dispose the sperm fluid when using the patient method, because the DNaseⅠ contained in the fluid can digest the exogenous DNA.
出处
《草食家畜》
2001年第S2期186-190,202,共6页
Grass-Feeding Livestock
关键词
精子载体介导
转基因
乳腺表达
药用蛋白质
sperm-mediated exogenous
DNA transfer
transgenic sheep
mammary gland expression
