摘要
本文以重组逆转录病毒衍生质粒的大肠杆菌工程菌种为材料,采用 Triton X-100裂解菌体,高速离心除去细菌染色体 DNA,稀酸沉淀除去蛋白质,以及 PEG 分部除去 RNA,结果获得与氯化铯-溴化乙锭密度梯度超速离心法的纯化效果相同的纯质粒制备物.本实验结果为基因克隆载体的制备提供了一个实用、简易和快速的方法.
An engineered E.coli strain harboring the plasmid derived from recombinant retroviral DNA
sequences was involved.Purified plasmids were obtained after lysing the bacteria by a detergent
Triton X-100,discarding bacterial cell debris and conjugated chromosomal DNAs by high-speed
centrifugation,deleting proteins by diluted acid precipitation,and suspending RNAs by
polyethylenglycol partition.It proved that the purification effect of this method was equivalent to
that of cesium chloride-ethidium bromide ultracentrifugation.It provided a practical,simple and
rapid procedure for preparing gene cloning vectors.
关键词
基因克隆
遗传工程
质粒
plasmid
gene clone
genetic engineering
