农业生物技术学报2011，Vol.19Issue(6)：1063-1069,7.DOI:10.3969/j.issn.1674-7968.2011.06.012
不同孵育时间对猪精子转染外源DNA效果的影响
The Influence of Incuabtion Time on the Transfection of Swine Sperm with Forigen DNA
摘要
Abstract
To obtain the optimism co-incubation time of boar sperm with DNA and set up the technique platform of swine sperm-mediated gene transfer, the Real time quantitative PCR, fluorescence microscope imaging and regular semen quality analysis were applied to detect the influence of incubation time of sperm with DNA on viability, motility, transfect efficiency, and the amount of adsorbed DNA and internalized DNA of sperm. The result revealed that, after 15, 30, 60, and 90 min co-incubation of the parceled and labeled DNA with sperm, the sperm motility and viability decreased extremely significantly with the increase of incubation time (P<0.0l), while the transfect efficiency increased extremely significantly (P<0.01) or significantly (P<0.05), as well as the amount of adsorbed DNA and internalized DNA increased, However, there was no difference among kicubation times (P>0.05) after 30 min incubation. In addition, compared with the increase of transfect efficiency of sperm, the decrease of motility and viability of sperm relatively was more obvious with the increase of co-incubation time. Overall, considering the parameters of motility, viability, transfect efficiency and the amount of adsorbed DNA and internalized DNA of sperm, the best co-incubation time of swine sperm with foreign DNA is 30 min.关键词
猪/精子/DNA/孵育时间Key words
Swine/ Sperm/ DNA/ Incubation time引用本文复制引用
宋成义,周辉云,谢雨琇,李庆平,高波,王宵燕,李碧春,毛九德..不同孵育时间对猪精子转染外源DNA效果的影响[J].农业生物技术学报,2011,19(6):1063-1069,7.基金项目
本研究由国家转基因生物新品种培育重大专项(No.2009ZX08010-019B、No.2008ZX08006-005)资助 （No.2009ZX08010-019B、No.2008ZX08006-005）
