title: Simultaneous Analysis of Multiple Promoters: An Application of the PC-GW Binary Vector Series.
abstract: With the advances in the field of synthetic biology, there is an increasing demand for multi-gene cloning technologies. Molecular cloning to generate multi-gene constructs can be performed by restriction digestion, or by recombination-based cloning strategies such as Gateway(®). This chapter details cloning, transformation, and selection procedures involved in generation of multi-gene expressing transgenic plants. Methods are described for cloning five distinct promoter-reporter fusion constructs into the PC-GW-BAR vector (from the PC-GW vector series) using Gateway(®) technology and meganuclease sites. Further, transformation and selection methods are described for the biofuel crop Camelina sativa from the Brassicaceae family. These methods would be constructive toward generating multi-gene expressing plants for simultaneous expression analysis of five promoters in a short time period.
authors.affiliation: Department of Crop Science, North Carolina State University, Campus Box 7287, Raleigh, NC, 27695, USA. fjyoti@ncsu.edu.
authors.forename: Jyoti
authors.initials: J
authors.lastname: Dalal
mesh_terms: D003001:Cloning, Molecular / Q000379:methods*; D018506:Gene Expression Regulation, Plant / Q000235:genetics; D005822:Genetic Vectors; D030821:Plants, Genetically Modified / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics*
keywords: Agrobacterium; Camelina; Gateway®; Meganuclease; Multi-gene cloning; PC-GW-BAR