title: A novel gateway-compatible binary vector series (PC-GW) for flexible cloning of multiple genes for genetic transformation of plants.
abstract: 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.
authors.affiliation: Crop Science Department, North Carolina State University, Raleigh, NC, USA. Electronic address: fjyoti@ncsu.edu.
authors.forename: Jyoti
authors.initials: J
authors.lastname: Dalal
authors.affiliation: Department of Plant and Microbial Biology, North Carolina State University, Raleigh, NC, USA. Electronic address: rdyalama@ncsu.edu.
authors.forename: Roopa
authors.initials: R
authors.lastname: Yalamanchili
authors.affiliation: Department of Plant and Microbial Biology, North Carolina State University, Raleigh, NC, USA. Electronic address: clahova@ncsu.edu.
authors.forename: Christophe
authors.initials: C
authors.lastname: La Hovary
authors.affiliation: Department of Plant and Microbial Biology, North Carolina State University, Raleigh, NC, USA. Electronic address: mikyoungjilee@gmail.com.
authors.forename: Mikyoung
authors.initials: M
authors.lastname: Ji
authors.affiliation: Department of Plant and Microbial Biology, North Carolina State University, Raleigh, NC, USA. Electronic address: mfrodri2@ncsu.edu.
authors.forename: Maria
authors.initials: M
authors.lastname: Rodriguez-Welsh
authors.affiliation: Department of Plant and Microbial Biology, North Carolina State University, Raleigh, NC, USA. Electronic address: dmaslett@ncsu.edu.
authors.forename: Denise
authors.initials: D
authors.lastname: Aslett
authors.affiliation: Crop Science Department, North Carolina State University, Raleigh, NC, USA. Electronic address: sganapa2@ncsu.edu.
authors.forename: Sowmya
authors.initials: S
authors.lastname: Ganapathy
authors.affiliation: Department of Plant and Microbial Biology, North Carolina State University, Raleigh, NC, USA. Electronic address: amgrunde@ncsu.edu.
authors.forename: Amy
authors.initials: A
authors.lastname: Grunden
authors.affiliation: Department of Plant and Microbial Biology, North Carolina State University, Raleigh, NC, USA. Electronic address: hwsedero@ncsu.edu.
authors.forename: Heike
authors.initials: H
authors.lastname: Sederoff
authors.affiliation: Crop Science Department, North Carolina State University, Raleigh, NC, USA. Electronic address: rongda_qu@ncsu.edu.
authors.forename: Rongda
authors.initials: R
authors.lastname: Qu
mesh_terms: D003001:Cloning, Molecular / Q000379:methods; D015870:Gene Expression; D023061:Gene Order; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics*; D030821:Plants, Genetically Modified; D010957:Plasmids / Q000235:genetics*; D014170:Transformation, Genetic; D019076:Transgenes
keywords: Agrobacterium; Gateway; Gene stacking; Plant transformation