A novel gateway-compatible binary vector series (PC-GW) for flexible cloning of multiple genes for genetic transformation of plants.
Jyoti Dalal, Roopa Yalamanchili, Christophe La Hovary, Mikyoung Ji, Maria Rodriguez-Welsh, Denise Aslett, Sowmya Ganapathy, Amy Grunden, Heike Sederoff, Rongda Qu
Author Information
Jyoti Dalal: Crop Science Department, North Carolina State University, Raleigh, NC, USA. Electronic address: fjyoti@ncsu.edu.
Roopa Yalamanchili: Department of Plant and Microbial Biology, North Carolina State University, Raleigh, NC, USA. Electronic address: rdyalama@ncsu.edu.
Christophe La Hovary: Department of Plant and Microbial Biology, North Carolina State University, Raleigh, NC, USA. Electronic address: clahova@ncsu.edu.
Mikyoung Ji: Department of Plant and Microbial Biology, North Carolina State University, Raleigh, NC, USA. Electronic address: mikyoungjilee@gmail.com.
Maria Rodriguez-Welsh: Department of Plant and Microbial Biology, North Carolina State University, Raleigh, NC, USA. Electronic address: mfrodri2@ncsu.edu.
Denise Aslett: Department of Plant and Microbial Biology, North Carolina State University, Raleigh, NC, USA. Electronic address: dmaslett@ncsu.edu.
Sowmya Ganapathy: Crop Science Department, North Carolina State University, Raleigh, NC, USA. Electronic address: sganapa2@ncsu.edu.
Amy Grunden: Department of Plant and Microbial Biology, North Carolina State University, Raleigh, NC, USA. Electronic address: amgrunde@ncsu.edu.
Heike Sederoff: Department of Plant and Microbial Biology, North Carolina State University, Raleigh, NC, USA. Electronic address: hwsedero@ncsu.edu.
Rongda Qu: Crop Science Department, North Carolina State University, Raleigh, NC, USA. Electronic address: rongda_qu@ncsu.edu.
The rapidly advancing field of plant synthetic biology requires transforming plants with multiple genes. This has sparked a growing interest in flexible plant transformation vectors, which can be used for multi-gene transformations. We have developed a novel binary vector series, named the PC-GW series (GenBank: KP826769-KP826773), for Agrobacterium-mediated plant transformation. The PC-GW vectors use the pCAMBIA vector backbone, and contain NPTII, hpt, bar, mCherry or egfp genes as selectable markers for plant transformation. In a modified multiple cloning site (MCS) of the T-DNA region, we have placed the attR1, attR2 and ccdB sequences for rapid cloning of one to four genes by Gateway™-assisted recombination. In addition, we have introduced four meganuclease sites, and other restriction sites for multi-gene vector construction. Finally, we have placed a CaMV 35S promoter and a 35S terminator on the 5' and 3' ends of the MCS. The CaMV 35S promoter is flanked by PstI restriction sites that can be used to replace it with another promoter sequence if needed. The PC-GW vectors provide choices for selectable markers, cloning methods, and can accommodate up to eight gene constructs in a single T-DNA, thereby significantly reducing the number of transformations or crosses needed to generate multi-transgene expressing plants.
