Summary
EcoRI fragments of the transducing phage λfus3 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 its 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 λrif d18 carrying tufB with DNA from pTUAl.
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References
Bachmann, B.J., Low, K.B., Taylor, A.L.: Recalibrated linkage map of Escherichia coli K-12. Bacteriol. Rev. 40, 116–167 (1976)
Blumenthal, R.M., Lemaux, P.G., Neidhardt, F.C., Dennis, P.P.: The effects of the relA gene on the synthesis of aminoacyl-tRNA synthetases and other transcription and translation proteins in Escherichia coli B. Mol. Gen. Genet. 149, 291–296 (1976)
Cameron, J.R., Panasenko, S.M., Lehman, I.R., Davis, R.W.: In vitro construction of bacteriophage λ carrying segments of the Escherichia coli chromosome: Selection of hybrids containing the gene for DNA ligase. Proc. Natl. Acad. Sci. U.S.A. 72, 3416–3420 (1975)
Dougan, G., Saul, M., Warren, G., Sherratt, D.: A functional map of plasmid ColEl. Mol. Gen. Genet. 158, 325–327 (1978)
Furano, A.V.: The elongation factor Tu coded by the tufA gene of Escherichia coli K-12 is almost identical to that coded by the tufB gene. J. Biol. Chem. 252, 2154–2157 (1977)
Furano, A.V.: Direct demonstration of duplicate tuf genes in enteric bacteria. Proc. Natl. Acad. Sci. U.S.A. 75, 3104–3108 (1978)
Furano, A.V., Wittel, F.P.: Syntheses of elongation factors Tu and G are under stringent control in Escherichia coli. J. Biol. Chem. 251, 898–901 (1976)
Gillespie, D., Spiegelman, S.: A quantitative assay for DNA-RNA hybrids with DNA immobilized on a membrane. J. Mol. Biol. 12, 829–842 (1965)
Gordon, J.: Regulation of the in vivo synthesis of the polypeptide chain elongation factors in Escherichia coli. Biochemistry 9, 912–917 (1970)
Jaskunas, S.R., Fallon, A.M., Nomura, M., Williams, B.G., Blattner, F.R.: Expression of ribosomal protein genes cloned in Charon vector phages and identification of their promoters. J. Biol. Chem. 252, 7355–7364 (1977)
Jaskunas, S.R., Lindahl, L., Nomura, M., Burgess, R.R.: Identification of two copies of the gene for the elongation factor EF-Tu in E. coli. Nature 257, 458–462 (1975)
Kaziro, Y.: The role of guanosine 5′-triphosphate in polypeptide chain elongation. Biochim. Biophys. Acta 505, 95–127 (1978)
Kirschbaum, J.B., Konrad, E.B.: Isolation of a specialized lambda transducing bacteriophage carrying the beta subunit gene for Escherichia coli ribonucleic acid polymerase. J. Bacteriol. 116, 517–526 (1973)
Lindahl, L., Post, L., Zengel, J., Gilbert, S.F., Strycharz, W.A., Nomura, M.: Mapping of ribosomal protein genes by in vitro protein synthesis using DNA fragments of λfus3 transducing phage DNA as templates. J. Biol. Chem. 252, 7365–7383 (1977)
Miller, D.L., Nagarkatti, S., Laursen, R.A., Parker, J., Friesen, J.D.: A comparison of the activities of the products of the two genes for elongation factor Tu. Mol. Gen. Genet. 159, 57–62 (1978)
Miller, D.L., Weissbach, H.: Factors involved in the transfer of aminoacyl-tRNA to the ribosome. In: Molecular mechanisms of protein biosynthesis (Weissbach, H., and Pestka, S., eds.), pp. 323–373. New York: Academic Press, 1977
Miller, J.H.: Experiments in molecular genetics. New York: Cold Spring Harbor Laboratory 1972
Miyajima, A., Kaziro, Y.: Coordination of level of elongation factors Tu, Ts, and G, and ribosomal protein Sl in Escherichia coli. J. Biochem. (Tokyo) 83, 453–462 (1978)
Nomura, M.: Some remarks on recent studies on the assembly of ribosomes. In: Nucleic acid-protein recognition (Vogel, H.J., ed.), pp. 443–467. New York: Academic Press 1977
Nomura, M., Morgan, E.A., Jaskunas, S.R.: Genetics of bacterial ribosomes. Annu. Rev. Genet. 11, 297–347 (1977)
Reeh, S., Pedersen, S., Friesen, J.D.: Biosynthetic regulation of individual proteins in relA + and relA strains of Escherichia coli during amino acid starvation. Mol. Gen. Genet. 149, 279–289 (1976)
So, M., Gill, R., Falkow, S.: The generation of a ColEl-Apr cloning vehicle which allows detection of inserted DNA. Mol. Gen. Genet. 142, 239–249 (1975)
Weber, K., Osborn, M.: The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J. Biol. Chem. 244, 4406–4412 (1969)
Zubay, G., Chambers, D.A., Cheong, L.C.: Cell-free studies on the regulation of the lac operon. In: The lactose operon (Beckwith, J.R., and Zipser, D., eds.), pp. 375–391. New York: Cold Spring Harbor Laboratory 1970
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Shibuya, M., Nashimoto, H. & Kaziro, Y. Cloning of an EcoRI fragment carrying E. coli tufA gene. Molec. gen. Genet. 170, 231–234 (1979). https://doi.org/10.1007/BF00337801
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DOI: https://doi.org/10.1007/BF00337801