Cloning of an EcoRI fragment carrying E. coli tufA gene

Abstract

EcoRI fragments of the transducing phage lambda fus3 DNA have been linked to the ColEl derivative plasmid RSF2124 (ColEl-Apr) DNA using bacteriophage T4 ligase. Among the plasmids formed, one designated pTUAl was found to contain the E. coli tufA gene. The proof for the presence of tufA gene in pTUAl is based on the following observations: (1) ability of pTUAl DNA and is EcoRI fragments to direct synthesis of EF-Tu in a cell-free protein synthesizing system; and (2) RNA . DNA hybridization of RNA transcribed from phage lambda rifd18 carrying tufB with DNA from pTUAl.