High Efficiency transformation ofSaccharomyces cerevisiaeby 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...
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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...
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This study will focus on how the different characteristics of the applied pulse, including voltage, pulse length, frequency and multiple pulsing affect transformation efficiency of Top10 E-coli cells. It will be shown that transformation efficiency will be maximal at optimum settings different from that suggested by electroporator manufacturer. Experimental results will also verify pore formati...
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Transformation efficiencies for Pichia pastoris are usually several orders of magnitude below those for other yeast. We report here that pretreatment of P. pastoris with 0.1 M lithium acetate (LiAc) and 10 mM dithiothreitol (DTT) before electroporation increased transformation efficiency approximately 150-fold. DTT alone enhanced the transformation efficiency up to 20-fold, but LiAc alone had l...
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ژورنال
عنوان ژورنال: Nucleic Acids Research
سال: 1993
ISSN: 0305-1048,1362-4962
DOI: 10.1093/nar/21.18.4414