实验动物与比较医学2013，Vol.33Issue(4)：256-260,5.DOI:10.3969/j.issn.1674-5817.2013.04.002
精子膜处理方法对精子介导的外源质粒转基因效率的影响
Impact of Different Method in Sperm Membrane Treatment on Exogenous Plasmid Transgenic Efficiency by Sperm
摘要
Abstract
Objective To explore the effect of three methods in removing mouse sperm membrane including ultrasound,NaOH and lysolecithin on the efficiency of transgenic by intracytoplasmic sperm injection (ICSI).Methods The mouse sperms were treated with the three methods above mentioned.Both of sperm membrane and DNA integrity of the sperm cells were evaluated by flow cytometry after died with Transgreen and propidium iodide (PI).The treated sperms were incubated with GFP-carrying plasmid and then conducted ICSI.The effect of these methods on efficiency of transgenic was evaluated by the results of fertilization cleavage and blastocyst with fluorescence.Results The rate of removing sperm membrane by lysolecithin method was higher than that of ultrasound and NaOH methods (99.2％ vs.94.7％,90.9％)(P＜0.05),but the rates of sperm DNA damage,the development of embryos from ICSI and blastocyst with fluorescence were similar among these methods (P＞0.05).Conclusion The three methods of mouse membrane treatment could be used to improve mouse transgenic efficiency by ICSI.关键词
精子去膜法/卵胞质内单精子注射(ICSI)/DNA完整性/转基因效率/小鼠Key words
Method/ Sperm membrane treatment/ ICSI/ DNA integrity/ Transgenic efficiency/ Mouse分类
生物科学引用本文复制引用
贾晓峰,徐彦,刘淼,王亮,施惠娟..精子膜处理方法对精子介导的外源质粒转基因效率的影响[J].实验动物与比较医学,2013,33(4):256-260,5.基金项目
上海市基础研究重点项目(10JC1409900)和上海市重点学科建设项目(S30201)资助 （10JC1409900）
