High-efficiency transformation of bacterial cells by electroporation
نویسندگان
چکیده
منابع مشابه
High efficiency transformation of E. coli by high voltage electroporation
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...
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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...
متن کاملHigh-efficiency transformation of Chlamydomonas reinhardtii by electroporation.
We have established a high-efficiency method for transforming the unicellular, green alga Chlamydomonas reinhardtii by electroporation. Electroporation of strains CC3395 and CC425, cell wall-less mutants devoid of argininosuccinate lyase (encoded by ARG7), in the presence of the plasmid pJD67 (which contains ARG7) was used to optimize conditions for the introduction of exogenous DNA. The condit...
متن کاملHigh-efficiency transformation of Saccharomyces cerevisiae cells by bacterial minicell protoplast fusion.
After a new transformation procedure, 10% of Saccharomyces cerevisiae cells were found to contain transforming DNA sequences. We used direct transfer of plasmid molecules by fusing bacterial minicell protoplasts to yeast protoplasts. Since the procedure significantly reduces the toxic effect of procaryotic protoplasm on the eucaryotic organism, it might be generally applicable in other systems ...
متن کاملImpact of heat shock step on bacterial transformation efficiency
CaCl2 treatment followed by heat shock is the most common method for artificial transformation. Here, the cells were transformed using CaCl2 treatment either with heat shock (standard protocol) or without heat shock (lab protocol) to comprehend the difference in transformation efficiency. The BL21 strain of Escherichia coli (E. coli) was being susceptible using CaCl2 treatment. Some Cells were ...
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ژورنال
عنوان ژورنال: Journal of Bacteriology
سال: 1988
ISSN: 0021-9193,1098-5530
DOI: 10.1128/jb.170.6.2796-2801.1988