Transient transformation is simpler, more efficient and economical in analyzing protein subcellular localization than stable transformation. Fluorescent fusion proteins were often used in transient transformation to follow the in vivo behavior of proteins. Onion epidermis, which has large, living and transparent cells in a monolayer, is suitable to visualize fluorescent fusion proteins. The oft...

The life cycle of Dictyostelium discoideum consists of many cellular and developmental aspects. By virtue of its relatively high transformation efficiency and a small haploid genome, this organism has proven to be advantageous for characterizing gene functions. However, a much higher transformation efficiency is required as one of the prerequisites for unraveling gene function on a genome-wide ...

Dictyostelium discoideum has been used as a genetically tractable model organism to study many biological phenomena. High-efficiency transformation is a prerequisite for successful genetic screens such as mutant complementation, identification of suppressor genes, or insertional mutagenesis. Although exponential decay electroporation is the standard transformation technique for D. discoideum, i...

Electrotransformation of Rhizobium leguminosarum was successfully carried out with a 15.1-kb plasmid, pMP154 (Cmr), containing a nodABC-lacZ fusion by electroporation. The maximum transformation efficiency, 10(8) transformants/microg of DNA, was achieved at a field strength of 14 kV/cm with a pulse of 7.3 ms (186 Omega). The number of transformants was found to increase with increasing cell den...

Escherichia coli as a plasmid recipient cell was dispersed in a chrysotile colloidal solution, containing chrysotile adsorbed to plasmid DNA (chrysotile-plasmid cell mixture). Following this, the chrysotile-plasmid cell mixture was dropped onto the surface of an elastic body, such as agarose, and treated physically by sliding a polystyrene streak bar over the elastic body to create friction. Pl...

We developed an improved protocol for the electrotransformation of Corynebacterium pseudotuberculosis, testing variations of parameters in the procedures that are routinely used for the preparation of electrocompetent cells of this species, including (i) culture conditions, (ii) cell growth phase, (iii) electroporation solutions and (iv) quantity of plasmid DNA. We obtained the greatest efficie...

An efficient Agrobacterium-mediate transformation method was developed by employing different duration of sonication, Agrobacterium strains, type of inoculation medium and concentrations of acetosyringone in Arya cultivar of wheat. Immature embryos were used as an explant for inoculation with Agrobacterium tumefaciens harboring the recombinant pBI121 plasmid. Among the durations of sonication t...

Genetic transformation studies were carried out to standardize a protocol for Agrobacterium -mediated genetic transformation of three economically important strawberry (Fragaria x ananassa Duch) cultivars Kurdistan’, Camarosa’, and ‘Paros’. Shoot regeneration frequency 72, 65 and 30% was obtained on MS (1) basal medium supplemented with 2% glucose and 4 mg/l TDZ for Camarosa, Kurdistan and Paro...

One model of transformation proposed that heat shock destabilizes the cell membrane and allows for plasmid uptake. As lipid composition might also affect the stability of the cell membrane, we hypothesized that a lower growth temperature would alter membrane lipid composition so as to increase transformation efficiency after heat shock. Cells were grown at 20°C and 37°C and treated with calcium...