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影响玉米基因枪转化效率的几个因素
Factors Influencing Maize Transformation by Particle Bombardments
【摘要】 通过检测GUS基因在玉米幼胚中的瞬时表达,研究了影响基因枪转化效率的几个因素。用质粒DNA包被金粉微粒,CaCl2溶液在pH为10与pH为5时的包被效果无明显差异,Ca(NO3)2溶液pH为10时优于pH为5时的包被效果。无论pH为5或10,CaCl2的包被效果均优于Ca(NO3)2。研究结果还证明,无亚精胺的包被效果与0.1mol·L-1亚精胺溶液基本相同;用高渗培养基处理幼胚能显著提高GUS基因的瞬时表达;GUS基因在不同的启动子调控下的瞬时表达效果明显不同,Actin1-GUS瞬时表达最强,Adh1-GUS居中,CaMV35S-GUS最弱。
【Abstract】 Factors influencing particle bombardment have been studied using the method of GUS transient expression.The results showed that 2.5 mol·L -1 Ca(NO 3) 2 at pH 10.0 gave a better result of GUS transient expression than that at pH 5.0; there is no obvious difference between pH 10.0 and pH 5.0 under 2.5 mol·L -1 CaCl 2 ; CaCl 2 was found to be superior to Ca(NO 3) 2 at both pH 10.0 and pH 5.0; No difference was found with or without spermidine in coating procedure.Osmotic treatment can improve the transient expression of GUS gene.Different levels of transient expression of GUS gene were found when driven by different promoters. The promoter derived from the rice actin1 gene produced significantly more GUS foci compared to the maize Adh1 and CaMV35S promoters.
【Key words】 maize immature embryos; particle bombardment; promoter; transient expression;
- 【文献出处】 农业生物技术学报 ,JOURNAL OF AGRICULTURAL BIOTECHNOLOGY , 编辑部邮箱 ,1997年01期
- 【分类号】Q78
- 【被引频次】40
- 【下载频次】374