Sedivy J M, Daldal F, Fraenkel D G
J Bacteriol. 1984 Jun;158(3):1048-53. doi: 10.1128/jb.158.3.1048-1053.1984.
The fbp locus at 96 min on the Escherichia coli chromosome governs fructose bisphosphatase (fructose-1,6-P2 1-phosphatase). We have cloned and subcloned fbp on vector pBR322 to obtain strains with high levels of the enzyme. In vivo mutagenesis of the clone was used to show that fbp is the structural gene. The gene was deleted on the plasmid in vitro, and the chromosomal wild-type locus was replaced with this deletion by a method involving stabilization of a heterozygous intermediate resulting from plasmid integration, followed by segregation of the wild-type gene.
大肠杆菌染色体上96分钟处的fbp基因座控制着果糖双磷酸酶（果糖-1,6-二磷酸1-磷酸酶）。我们已将fbp克隆并亚克隆到载体pBR322上，以获得具有高水平该酶的菌株。利用该克隆的体内诱变来表明fbp是结构基因。该基因在体外质粒上被删除，并且通过一种涉及稳定由质粒整合产生的杂合中间体，然后分离野生型基因的方法，用这种缺失取代了染色体野生型基因座。
