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| The Influence of Incuabtion Time on the Transfection of Swine Sperm with Forigen DNA |
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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<0.01), 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 incubation 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.
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Received: 21 June 2011
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ZHU Xiao-Feng, XU Hou-Qiang, CHEN Wei, CHEN Ying-Lian, NI Kai, WU Xiao-Min, NI Meng-Meng, LU Xian-Jun. Cloning, Expression and Bioinformatics Analysis of IGF-1 and IGF-2 Genes in Congjiangxiang Pigs (Sus scrofa)[J]. 农业生物技术学报, 2019, 27(8): 1382-1391. |
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