Li J T, Yu G, Sun X H, Jia C G, Du Q, Li Q Y, Pan H Y
College of Plant Science, Jilin University, Changchun, China.
Jilin Academy of Agricultural Science, Changchun, China.
Genet Mol Res. 2014 Sep 26;13(3):7815-25. doi: 10.4238/2014.September.26.20.
Simple, efficient, and economical recombinant plant binary expression vectors for deciphering large-scale functional genomic research in plants and promoting crop improvement by genetically engineering and biotechnology is in great demand. In this research, using the pCHF3, pCAMBIA1301, pCAMBIA3300, pCAMBIA3301 vectors, we successfully constructed general plant binary expression vectors carrying CaMV35S and Arabidopsis rd29A promoters mediating multiple cloning sites (
SacI, KpnI, SmaI, BamHI, XbaI, SalI, and PstI). Meanwhile, a series of applicative binary expression vectors that can be utilized for subcellular localization were constructed by fusion of the MCS and eGFP. Subsequently, the recombinant vectors were successfully transferred into Arabidopsis thaliana and Nicotiana benthamiana for further investigation of functional elements in these plant binary expression vectors. Our results demonstrated that this system was a convenient and versatile vector system for phenotypic, functional, subcellular localization, and promoter activity analysis, and it provided a relatively high-efficiency and reliable platform for researchers in vector construction and may facilitate large-scale functional genomics analysis in plants.
用于解析植物大规模功能基因组研究以及通过基因工程和生物技术促进作物改良的简单、高效且经济的重组植物双元表达载体需求巨大。在本研究中，我们使用pCHF3、pCAMBIA1301、pCAMBIA3300、pCAMBIA3301载体，成功构建了携带CaMV35S和拟南芥rd29A启动子并介导多克隆位点（MCS：SacI、KpnI、SmaI、BamHI、XbaI、SalI和PstI）的通用植物双元表达载体。同时，通过将MCS与eGFP融合构建了一系列可用于亚细胞定位的应用双元表达载体。随后，将重组载体成功转入拟南芥和本氏烟草，以进一步研究这些植物双元表达载体中的功能元件。我们的结果表明，该系统是用于表型、功能、亚细胞定位和启动子活性分析的便捷且通用的载体系统，为研究人员进行载体构建提供了一个相对高效且可靠的平台，并可能促进植物大规模功能基因组学分析。
