质粒
转座因子
转座子突变
穿梭机载体
生物
大肠杆菌
复制子
突变
多克隆站点
睡美人转座系统
插入顺序
基因组
遗传学
基因
载体(分子生物学)
突变
重组DNA
作者
Reinhard Simon,Ursula B. Priefer,Alfred Pühler
出处
期刊:Bio/technology
[Springer Nature]
日期:1983-11-01
卷期号:1 (9): 784-791
被引量:6731
摘要
We have developed a new vector strategy for the insertion of foreign genes into the genomes of gram negative bacteria not closely related to Escherichia coli. The system consists of two components: special E. coli donor strains and derivatives of E. coli vector plasmids. The donor strains (called mobilizing strains) carry the transfer genes of the broad host range IncP–type plasmid RP4 integrated in their chromosomes. They can utilize any gram negative bacterium as a recipient for conjugative DNA transfer. The vector plasmids contain the P–type specific recognition site for mobilization (Mob site) and can be mobilized with high frequency from the donor strains. The mobilizable vectors are derived from the commonly used E. coli vectors pACYC184, pACYC177, and pBR325, and are unable to replicate in strains outside the enteric bacterial group. Therefore, they are widely applicable as transposon carrier replicons for random transposon insertion mutagenesis in any strain into which they can be mobilized but not stably maintained. The vectors are especially useful for site–directed transposon mutagenesis and for site–specific gene transfer in a wide variety of gram negative organisms.
科研通智能强力驱动
Strongly Powered by AbleSci AI