Berthier Françoise, Zagorec Monique, Champomier-Vergès Marie, Ehrlich S D, Morel-Deville Françoise
Laboratoire de Recherches sur la Viande, Institut National de la Recherche Agronomique, Domaine de Vilvert, 78352 Jouy-en-Josas Cedex, France.
Laboratoire de Génétique Microbienne, Institut National de la Recherche Agronomique, Domaine de Vilvert, 78352 Jouy-en-Josas Cedex, France.
Microbiology (Reading). 1996 May;142(5):1273-1279. doi: 10.1099/13500872-142-5-1273.
A procedure to transform intact cells by electroporation was developed through a systematic examination of the effect of changes in various parameters on the transformation efficiency of strain 64F. The most critical factors were found to be the electrical parameters, the composition of washing and electroporation/storage solutions, and the presence of MgCI in the expression medium. Under optimal conditions transformation efficiencies up to 10 transformants (μg supercoiled DNA) were obtained. The optimized procedure was successfully applied to other strains and consistently yielded from 10 to 10 transformants (μg supercoiled DNA).
通过系统研究各种参数变化对64F菌株转化效率的影响，开发了一种通过电穿孔转化完整细胞的方法。发现最关键的因素是电参数、洗涤和电穿孔/储存溶液的组成以及表达培养基中MgCI的存在。在最佳条件下，获得了高达10个转化子/（μg超螺旋DNA）的转化效率。优化后的方法成功应用于其他菌株，始终产生10至10个转化子/（μg超螺旋DNA）。
