Article Abstract:
The L41 protein gene of the fermenting yeast Phaffia rhodozyma was cloned and then used as a cycloheximide resistance marker during the transformation of the yeast. Transformation efficiencies of 800 to 1,200 transformants per microgram of DNA were obtained. Expression of the cycloheximide resistance gene was achieved through postincubation of electroporated cells for 14 to 16 hours. Homologous recombination into the rDNA locus appeared to be the course by which plasmid integration was accomplished in most transformants.
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Article Abstract:
The conditions required for optimal electroporation efficiency in Gram-negative bacteria were determined. Experiments carried out on three species of Gram-negative bacteria showed that resistance of the bacterial suspensions fluctuates by four orders of magnitude during capacitor discharge. Furthermore, independent manipulations of pulse duration and capacitor time constants revealed that electrical energy delivered in each pulse is the major factor determining electrotransformation efficiency.
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Article Abstract:
Free DNA (lysates) or live donor cells naturally transform strains of the Acinetobacter calcoaceticus, incorporated in situ in river epilithon, to prototrophy. The age of the recipient indigenous community affects the transfer frequency in a laboratory setting. Though the transfer frequency is positively related with the temperature, river Taff below 10 degrees celsius fail to show any transformation. However, laboratory microorganisms are unable to reflect the actual environmental conditions.
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