影响玉米基因枪转化效率的几个因素  

Factors Influencing Maize Transformation by Particle Bombardments

作　　者：赵天永[1] 黄忠[1] 王国英[1] 谢友菊 

机构地区：[1]中国农业大学生物学院

出　　处：《农业生物技术学报》1997年第1期35-39,共5页Journal of Agricultural Biotechnology

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。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.

关 键 词：玉米 幼胚 基因枪 启动子 表达 

分 类 号：S513.035.3[农业科学—作物学]