Improved binary vectors forAgrobacterium-mediated plant transformation

✍ Scribed by Kevin E. McBride; Kristin R. Summerfelt

• Publisher: Springer
• Year: 1990
• Tongue: English
• Weight: 813 KB
• Volume: 14
• Category: Article
• ISSN: 0167-4412
• DOI: 10.1007/bf00018567

✦ Synopsis

Improved plant transformation vectors were constructed which utilize the pRiHRI origin of replication for highly stable maintenance in Agrobacterium tumefaciens, the ColE1 origin of replication for high copy maintenance in Escherichia coli, and a gentamycin resistance gene as a strong selectable marker for bacteria. Concise T -D N A elements were engineered with border sequences from the T L -D N A of pTiA6, the Tn5 neomycin phosphotransferase gene (npt H) expressed from either CaMV 35S or mannopine synthase (mas) promoters, and the lac Z' gene segment from pUC18 as a source of unique restriction sites as well as an insertional inactivation marker for cloned DNA. The order of T-DNA components in all vectors is left border, plant marker cassette, lac Z', and right border, respectively. The prototype vector, pCGN1547, was shown to be very stable in A. tumefaciens strain LBA4404 and to act as an efficient donor of T-DNA in tomato transformation experiments. Use of the other vectors is also described.