A rapid procedure for cloning genes from lambda libraries by complementation of E. coli defective mutants: application to the fabE region of the E. coli chromosome.

J H Alix
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引用次数: 22

Abstract

I describe a general and rapid procedure allowing the isolation of specialized lambda transducing phages from a lambda library by lysogenic complementation of defective mutants of Escherichia coli. As an example, the cloning of the E. coli fabE gene and of two other adjacent genetic determinants is presented. Subcloning and determination of its nucleotide sequence reveals that fabE codes for the biotin carboxyl carrier protein (BCCP), one of the three subunits of acetyl coenzyme A carboxylase.

利用大肠杆菌缺陷突变体的互补从λ文库中快速克隆基因的方法:应用于大肠杆菌染色体fabE区。
我描述了一种通用和快速的程序,允许通过溶原性互补的大肠杆菌缺陷突变体从lambda文库中分离出专门的lambda转导噬菌体。作为一个例子,克隆大肠杆菌fabE基因和其他两个邻近的遗传决定因素提出。亚克隆和核苷酸序列测定表明,fabE编码生物素羧基载体蛋白(BCCP),是乙酰辅酶A羧化酶的三个亚基之一。
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