家畜生态学报2012，Vol.33Issue(1)：23-28,6.
影响猪精子介导基因转移效果的因素研究
Factors Affecting Porcine Sperm Mediated Gene Transfer
摘要
Abstract
To optimize the technical parameters of porcine sperm mediated gene transfer, the real-time PCR, fluorescence microscope imaging and regular semen quality analysis were applied to detect the influence of DNA transfect reagent, incubation temperature and DNA form on the transfection of porcine sperm with foreign DNA. The result revealed that PEI and TransFast significantly improved the transfection of porcine sperm with DNA, and PEI was better than TransFast, but NanoFect blocked the transfection completely. With the increase of incubation temperature, the viability, motility, and the amount of binding DNA of sperm decreased significantly, while the transfection efficiency and amount of internalized DNA were not affected. The difference of viability, motility, and amount of internalized DNA was not significant between circular and linear DNA incubated sperm, whereas the amount of binding DNA of sperm incubated with linear DNA was extremely higher than that of circular DNA. It could be concluded that the form of DNA had no significant influence on the transfection of porcine sperm, PEI and TransFast improved the transfection of porcine sperm significantly, and the best incubation temperature for porcine sperm is 17 ℃. This study provided basic data for further porcine transgenesis research with sperm mediated gene transfer.关键词
DNA转染剂/孵育温度/DNA形态/猪/精子Key words
DNA transfect reagent/incubation temperature/DNA form/porcine/sperm分类
农业科技引用本文复制引用
宋成义,吴晗,周辉云,谢雨琇,李庆平,高波,王宵燕,李碧春,毛九德..影响猪精子介导基因转移效果的因素研究[J].家畜生态学报,2012,33(1):23-28,6.基金项目
国家转基因生物新品种培育重大专项(2009ZX08010-019B （）
2008ZX08006-005) （）
