pORE: a modular binary vector series suited for both monocot and dicot plant transformation.
We present a series of 14 binary vectors suitable for Agrobacterium-mediated transformation of dicotyledonous plants and adaptable for biolistic transformation of monocotyledonous plants. The vector size has been minimized by eliminating all non-essential elements from the vector backbone and T-DNA regions while maintaining the ability to replicate independently. The smallest of the vector series is 6.3 kb and possesses an extensive multiple cloning site with 21 unique restriction endonuclease sites that are compatible with common cloning, protein expression, yeast two-hybrid and other binary vectors. The T-DNA region was engineered using a synthetic designer oligonucleotide resulting in an entirely modular system whereby any vector element can be independently exchanged. The high copy number ColE1 origin of replication has been included to enhance plasmid yield in Escherichia coli. FRT recombination sites flank the selectable marker cassette regions and allow for in planta excision by FLP recombinase. The pORE series consists of three basic types; an 'open' set for general plant transformation, a 'reporter' set for promoter analysis and an 'expression' set for constitutive expression of transgenes. The sets comprise various combinations of promoters (P (HPL), P (ENTCUP2) and P (TAPADH)), selectable markers (nptII and pat) and reporter genes (gusA and smgfp).
Coutu
Catherine
C
Agriculture and Agrifood Canada, Saskatoon Research Centre, 107 Science Place, Saskatoon, SK, Canada S7N 0X2.
Brandle
James
J
Brown
Dan
D
Brown
Kirk
K
Miki
Brian
B
Simmonds
John
J
Hegedus
Dwayne D
DD
D016960:Agrobacterium tumefaciens / Q000235:genetics; D017360:Arabidopsis / Q000235:genetics; D029688:Brassica napus / Q000235:genetics; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D018532:Pisum sativum / Q000235:genetics; D030821:Plants, Genetically Modified; D010957:Plasmids / Q000235:genetics*; D013025:Glycine max / Q000235:genetics; D014170:Transformation, Genetic; D003313:Zea mays / Q000235:genetics