By Kevan M. A. Gartland, Michael R. Davey
Agrobacterium Protocols deals starting and skilled researchers the main finished number of step by step protocols for the genetic manipulation of vegetation utilizing Agrobacterium. the subjects diversity from the upkeep of bacterial tradition collections to facets of the metabolism and body structure of reworked tissues and transgenic crops. Drawing at the paintings of best scientists from laboratories worldwide, Agrobacterium Protocols presents a wealth of concepts for introducing particular DNA sequences into goal plant species and discusses the environmental implications of genetically engineered crops. Its precise tactics will facilitate speedy move of complicated thoughts to different laboratories and their exploitation in basic and utilized plant biology.
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Extra info for Agrobacterium Protocols (Methods in Molecular Biology Vol 44)
7. 8: 500 mL acidic ATx2 stock solution, 1 g ammonium sulfate, 2 g glucose. Final vol, 1 L. 8. 1 mg acetosyringone in 1 mL dimethyl sulfoxide (DMSO). Keep at -20°C. 2. Recovery of Bacterial Clones Following Growth in the Presence of Acetosyringone and Recognition of Avirulent Mutants 1. Nopaline stock solution: Dissolve 800 mg nopaline (Sigma, St. Louis, MO) in 100 mL water. The addition of l-2 drops of 6N KOH may be necessary to facilitate solubilization. Sterilize by filtration. Only nopaline-type strains have so far been found to be susceptible to acetosyringone-induced growth inhibition.
The binary system is much easier to use since binary vectors are smaller (about lo-15 kb) and do not require cointegration. After insertion of a target gene into a unique site in the T-DNA region using E. coli as a host, the vector is transferred into Agrobacterium by either conjugation or DNA transformation. As the binary vector does not carry the mobilization function necessaryfor conjugal transfer, this function is provided by a mobilization plasmid carried in another E. coli strain. The advantage of the conjugal transfer method is a higher transformation frequency compared to the direct DNA transformation.
B. (1992) Non-destructive assay systems for detection of P-glucuronidase activity in higher plants Plant Mol Biol. Rep. 10,37-46. 22. , and Goldberg, R. B. (1990) Introduction of male sterility m plants by a chimerlc rlbonuclease gene Nature 347,737-74 1. 23 Green, P. , and Inouye, M. (1986) The role of antisense RNA m gene regulation. Annu Rev. Blochem. 55,569-597. 24. Oeller, P. , Taylor, L. , Pike, D. A , and Theologis, A. (199 1) Reversible inhibition of tomato fruit senescence by antisense RNA.
Agrobacterium Protocols (Methods in Molecular Biology Vol 44) by Kevan M. A. Gartland, Michael R. Davey