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URL: http://www.addgene.org/11570
Proper Citation: RRID:Addgene_11570
Insert Name: CaMV 35S enhancers
Organism: Cauliflower mosaic virus
Bacterial Resistance: Ampicillin
Defining Citation: PMID:10759496
Vector Backbone Description: Backbone Marker:Hayashi et al, 1992; Backbone Size:10138; Vector Backbone:pPCVICEn4HPT; Vector Types:Plant Expression, Activation tagging; Bacterial Resistance:Ampicillin
Comments: The CaMV 35S enhancers in these vectors correspond to nucleotides -417 to -86 relative to the transcription start. Recommended Agrobacterium strain is GV3101 pMP90RK. Host and helper plasmid markers are kanamycin, gentamycin and rifampicin resistance. Plasmid selection in E. coli and in A. tumefaciens is ampicillin resistance. Sites that leave the pBstKS+ sequences intact and cut only on one side between pBstKS+ and either left or right border can be used for plasmid rescue. Note that there are additional BssS1 sites not shown on the map. The four enhancer repeats in the construct may be unstable in E. coli and Agrobacterium if stored at +4'C for extended time. Check by PCR with T7 (or M13-20) oligo and this one derived from the RB sequence: 5' acc cgc caa tat atc ctg 3'. This should give 1.4 kb band. Note that these oligos will not work in a transgenic plant, as the RB sequence is not transfered. Using this test, we found that after 1-2 weeks in a fridge the Agrobacterium strain loses on average one copy of a repeat, and after a month in a fridge there is only one copy left. It is a good idea to use a freshly streaked colony from a good -70'C glycerol stock for every infiltration.
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Source: Addgene
