4.1 Article

Modification of vectors for functional genomic analysis in plants

Journal

GENETICS AND MOLECULAR RESEARCH
Volume 13, Issue 3, Pages 7815-7825

Publisher

FUNPEC-EDITORA
DOI: 10.4238/2014.September.26.20

Keywords

Plant binary expression vector; Stable expression; Stress-inducible expression; Subcellular localization

Funding

  1. National Science and Technology Support Program [2012BAD19B04]
  2. Ministry of Agriculture Key Projects of GM Cultivation of New Varieties [2013ZX08004004]

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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 (MCS: 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.

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