pKYLX5 BioVector NTCC质粒载体菌种细胞基因保藏中心

pKYLX5

We have assembled a series of expression vectors for use with Agrobacterium-mediated and biolostic gene transfer in plants. The Agrobacterium-based plasmids (pKYLX5, pKYLX7, pKYLX71, and pKYLX71:35S2) all use, as a "shell", the pGA472 plasmid described by An et al. (1); the construction and basic characterization of pKYLX5 and pKYLX7 have been described (2). The biolistic plasmid (pKYLX80) contains the expression cassette and kanamycin resistance gene from pKYLX71:35S2, but in a slightly modified pBluescript background; this permits the production of large quantities of these plasmids.
Key: Restriction enzyme sites are as shown.
TL, TR - left and right T-DNA borders (see reference 2 for a detailed description of their origin).
The multiple cloning site for pKYLX71 and pKYLX71:35S2 (these are the plasmids currently in use) is: HindIII*-BamHI-XhoI*-PstI-SacI*-XbaI* (* - unique sites).
The promoters that have been described are:

in pKYLX5 - the pea rbcS-E9 promoter
in pKYLX7 and pKYLX71 - the CaMV 35S promoter
in pKYLX71:35S2 - a modified 35S promoter with a duplicated "enhancer" region

Arrows show directions of transcription for the expression cassette and kanamycin resistance gene.
This plasmid confers tetracycline (12.5 µg/ml) and kanamycin (50 µg/ml) resistance upon E. coli and Agrobacterium tumefaciens. The kanamycin resistance gene is effective at kanamycin concentrations as high as 500 µg/ml in tobacco. These plasmids have been used in several labs, and work well in Arabidopsis.

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