To develop a gene transformation method for Flammulina velutipes, we constructed a vector with hph gene under control of the trp1 gene promoter. The vector was integrated into protoplast derived from mycelia by the calcium-polyethylene glycol method, as it has not been reported for F. velutipes. Transformation efficiency was much improved when transformation was performed by the restriction enz...

With ever shorter development times and market demands on overall system performance such as efficiency, reliability and low maintenance, accurate predictive tools are necessary and gear drives prove to be no exception. All these characteristics have an impact on a process which has remained a riddle: running-in. Even though no consensus on a definition of this phenomena is readily available, t...

We describe a simple calcium phosphate transfection protocol and neo marker vectors that achieve highly efficient transformation of mammalian cells. In this protocol, the calcium phosphate-DNA complex is formed gradually in the medium during incubation with cells and precipitates on the cells. The crucial factors for obtaining efficient transformation are the pH (6.95) of the buffer used for th...

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We report a simple, rapid, 5 min transformation protocol that produces ∼100% more transformants than the original calcium competent protocol of Mandel and Higa (1). There have been improvements on the transformation efficiency of 105–106 colonies/μg plasmid DNA (2); the best probably that of Hanahan (3) yielding 107–109. The highest rates are achieved using electroporation (109–1010). However, ...

Streptococcus lactis IL1403 protoplasts were transformed by plasmid pIL204 (5.5 kilobases), which conferred erythromycin resistance with an average efficiency of 5 X 10(6) transformants per microgram of supercoiled DNA. The procedure used and transformation efficiencies obtained were close to those described for Bacillus subtilis (G. Chang and S. N. Cohen, Mol. Gen. Genet. 168:111-115, 1979).

E. coli can be transformed to extremely high efficiencies by subjecting a mixture of cells and DNA to brief but intense electrical fields of exponential decay waveform (electroporation). We have obtained 10(9) to 10(10) transformants/micrograms with strains LE392 and DH5 alpha, and plasmids pUC18 and pBR329. The process is highly dependent on two characteristics of the electrical pulse: the ele...

Several methods of transformation are currently available for delivering exogenous DNA to plant cells. Agrobacterium-mediated transformation, microprojectile bombardment and direct protoplast transformation are routinely used today. However, each of them has certain disadvantages, which led to research into the development of novel alternative systems such as infiltration, electroporation of ce...

In this article, we develop the Sinc-Galerkin method based on double exponential transformation for solving a class of weakly singular nonlinear two-point boundary value problems with nonhomogeneous boundary conditions. Also several examples are solved to show the accuracy efficiency of the presented method. We compare the obtained numerical results with results of the other existing methods in...