摘要
本实验的主要目的旨在摸索电场对基因转化的最佳条件.已得到初步结果为:用酿酒酵母S.cerevisiae DBY 746作为穿梭质粒(YRp类)的受体菌.经过对最适电场条件与基因转化率之间关系的研究,我们发现在1个400μS的宽时程脉冲、电场强度为4KV/cm时有最高转化率,达273个转化子/μgDNA.本实验所用的电穿孔装置是自组装的,它简便、快速、实用.
The electroporation technology is a bioelectric new techniques which has been gradually in gene transformation in recent years. It makes outside gene be led into cell and expressed by pore effect under a D.C. high vol-tage electric pulse to biologic mambrane. This experimental key purpose lies in tryine to find out the optimal condition of electric field to gene transformation. Now the initial results obtained under this experiment condition were: Shuttle plasmid (YRp type) to Sacchararomyces cerevisiae transfered, the maxium of efficiecy of transformation was observed, it reached about 273 transformants per ?g DNA at 4KV/ cm (electric field strength), 400μs(pulse time constant) and a unique pulse. Meanwhile, it was found that plasmid efficiency of transformation and yeast proto-plastsviability all became lower by the number of pulse increasing. This experiment adopted the wide pulse (400μs) for transformation yeast protoplasts, it hasn't been reported home and abroad.
出处
《生物物理学报》
CAS
CSCD
北大核心
1991年第3期391-397,共7页
Acta Biophysica Sinica
关键词
电穿孔技术
质粒
酵母
原生质体
Electroporation
Saccharomyces Cerevisiae
Protoplasts
Plasmid
Transformation.
