摘要
分别以洋葱表皮和生长7d的小麦连体叶片为材料,采用HeliosTM基因枪将35S-sGFP-TYG-NOS(pUC19)外源基因导入植物细胞使其瞬时表达,摸索出一整套适合于小麦连体叶片高效瞬时表达体系的各项参数,包括:氦气压力、金粒子直径、PVP浓度、每次轰击的质粒DNA以及金粉用量等。并指出选用合适的质粒DNA浓度和多片小麦叶片同时轰击可有效提高转化效率。
The Helios^TM Gene Gun is the second instrument in Bio-Rad＇s particle delivery product line. It provides a much wider selection of target materials. The aim of this study was to optimize parameters for this equipment to transiently express of gene constructs in intact wheat leaves. In order to investigate the efficacy of transient gene delivery, we used constructs containing green fluorescent protein gene that were driven by the constitutively active 35S promoter of the CaMV 35S gene, with as a reporter. Parameters optimized included helium pressure, the size of gold particles, the concentration of PVP, DNA amount and the amount of gold per shot. We found that optimal amount of plasmid DNA plays a key role in the efficiency of transient expression. The possible measures to improve the consistency and the reliability of transient gene expression were presented.
出处
《中国农学通报》
CSCD
2007年第7期280-284,共5页
Chinese Agricultural Science Bulletin
基金
973"国家重点基础研究项目"主要农作物核心种质重要功能基因多样性及其应用价值研究"(2003AA207160)资助
