生命科学研究2018，Vol.22Issue(2)：114-121,8.DOI:10.16605/j.cnki.1007-7847.2018.02.004
用于植物基因表达载体构建的质粒改造及其应用
Plasmid Modification for Construction of Plant Gene Expression Vector and Its Applications
摘要
Abstract
In order to overcome the defects that the commonly used plasmids have a limited number of restriction sites for target gene cloning, and lack expressing elements such as promoter, terminator and selection marker genes, a plasmid named pNULPGE200 for construction of plant gene expression vector was modified. The plasmid contains cauliflower mosaic virus (CaMV) 35S promoter, nopaline synthase (NOS) ter-minator, and a multiple cloning site between them. The target gene amplified by PCR can be inserted directly between the 35S promoter and the NOS terminator, and can be expressed in plants stably. pNULPGE200 also contains two independent expression marker genes, encoding neomycin phosphotransferase Ⅱ(NPT Ⅱ) and synthetic green-fluorescent protein with S65T mutation (sGFP), which can be used for mutual selection in plant transformation. The practicability of the plasmid was confirmed by comparing with a conventional plas-mid pBI121 in the construction of the gene encoding xylene monooxygenase from Pseudomonas putida to create a plant gene expression vector.关键词
质粒改造/植物基因表达载体/载体构建/二甲苯单加氧酶基因Key words
plasmid modification/plant gene expression vector/vector construction/xylene monooxygenase gene分类
生物科学引用本文复制引用
安韶雅,虎娟,张虹,孙放,马霞,王晨,陈任..用于植物基因表达载体构建的质粒改造及其应用[J].生命科学研究,2018,22(2):114-121,8.基金项目
宁夏大学引进人才研启动基金资助项目(BQC2012001) （BQC2012001）
国家自然科学基金项目(31460062) （31460062）
宁夏大学创新实验项目(X2016037) （X2016037）
