By Kevan M. A. Gartland, Michael R. Davey
Agrobacterium Protocols bargains starting and skilled researchers the main finished choice of step by step protocols for the genetic manipulation of crops utilizing Agrobacterium. the subjects diversity from the upkeep of bacterial tradition collections to points of the metabolism and body structure of reworked tissues and transgenic vegetation. Drawing at the paintings of prime scientists from laboratories worldwide, Agrobacterium Protocols offers a wealth of innovations for introducing particular DNA sequences into aim plant species and discusses the environmental implications of genetically engineered crops. Its designated tactics will facilitate speedy move of complex innovations to different laboratories and their exploitation in primary and utilized plant biology.
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Extra resources for Agrobacterium Protocols (Methods in Molecular Biology Vol 44)
8. Stachel, S. E and Nester, E W. (1986) The genetic and transcriptional organization of the vir region of the A6 TI plasmid of Agrobacterium tumefaciens. EMBO J. $1445-1454 9. Stachel, S E and Zambryski, P (1986) virA and virG control the plant-induced activation of the T-DNA transfer process of Agrobacterium tumefaciens. Cell 46,325-333 10. Douglas, C. , and Nester, E. W. (1982)Agrobacterium tumefaciens mutants affected m attachment to plant cells. J. Bacterial. 152, 1265-1275. 11. An, G , Ebert, P.
6, Count colonies after 2-5 d incubation at 28OC. 7. Perform assaysin triplicate. 5. Blindwell Assays Early experiments were performed using the capillary assay (13,14). We have subsequently switched to the Blindwell assay (5) owing to its ease of use and greater reproducibility. These are performed in Boyden chambers (15), which consist of a perspex base, drilled and threaded, into which a hollow Teflon plug is screwed (see Fig. 2). 1. Lightly grease shelf above the bottom chamber. 2. tL of bacteria into the bottom chamber.
K (1985) Resistance to hygromycin B A new marker for plant transformation studres. Plant Mol Biol 5, 103-108. 6 Holt, J. , Powles, S. , and Holtum, A M. (1993) Mechanisms and agronomic aspect of herbicide resistance. Annu. Rev. Plant Physiol. Plant Mol. Biol. 44,203-229. , Hirsch, P. , Hooykaas, P J J , and Schilperoort, R. A (1983) A binary plant vector strategy based on separation of vir- and T-region of the Agrobacterium tumefaciens Tr-plasmrd. Nature 303, 179,180. 8. Stachel, S. E and Nester, E W.