Mapping of ribosomal protein genes by in vitro protein synthesis using DNA fragments of lambda fus3 transducing phage DNA as templates

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dc.contributor.authorLindahl, Lasse
dc.contributor.authorPost, Leonard
dc.contributor.authorZengel, Janice
dc.contributor.authorGilbert, Scott F.
dc.contributor.authorStrycharz, William A.
dc.contributor.authorNomura, Masayasu
dc.date.accessioned2019-07-01T15:05:50Z
dc.date.available2019-07-01T15:05:50Z
dc.date.issued1977-10-25
dc.description.abstracthfus3 (or hfus2) transducing phage stimulates the synthesis of 27 ribosomal proteins, elongation factors EF-Tu and EF-G, and RNA polymerase subunit (Y in ultraviolet-irradiated Escherichia roli cells (see a preceding paper, Jaskunas, S. R., Fallon, A. M., and Nomura M. (1977) .I. Bid. Chem. 252, 7323-7336). The presence of the structural genes for these proteins on the hfus3 genome was proven by demonstrating the synthesis of these proteins in a DNA-dependent protein-synthesizing system using Afus3 DNA, or its restriction enzyme fragments, or both, as template. Proteins synthesized in the system were identified by a combination of techniques. including gel electrophoresis, immunochemical reactions, and tryptic peptide analysis. Restriction endonuclease-sensitive sites on kfus3 DNA were mapped, and genes were located using restriction endonuclease fragments of Afus3 DNA, isolated directly or cloned in phage or plasmid vectors, as templates for protein synthesis. Additional information about the locations of the genes was obtained using DNA from insertion and deletion mutants of hfux3. The results obtained in these in oilro experiments (rf. Fig. 2), and those obtained in the in uiuo experiments from preceding papers in this series, are in excellent agreement and map the essential genes on Afun and their promoters as follows (going counter-clockwise on the E. coli chromosome): R,,.. (S12, S7), EF-G, EF-Tu; k,,), SlO, LB, (1~4. L2. L23), (L22. S19), s3, (S17, L16, L29); hUC, L14, L34, L5, S14, S8, L6, LlR, S5, (130, L15); pa, S13, Sll, S4, IY, L17. Re-examination of the results of previous physiological experiments (Dennis, P. P. (1974) J. Mol. Biol. 89, 223- 232) using the present information on r-protein gene organization demonstrates that the increase in the differential synthesis rate of ribosomal proteins during the transition period following the nutritional shift-up is primarily due to the increase in the frequency of initiation of the transcription of ribosomal protein genes.en_US
dc.description.sponsorshipThis work was supported in part by the college of Agriculture and Life Sciences, University of Wisconsin, and by grants from the National Institute of General Medical Sciences (GM-20427) and the National Science Foundation (GB-31086).en_US
dc.description.urihttps://pdfs.semanticscholar.org/8d0a/ef8155e29501954fafa585271e3ad3991b25.pdfen_US
dc.format.extent19 pagesen_US
dc.genrejournal articlesen_US
dc.identifierdoi:10.13016/m2geuo-1cia
dc.identifier.citationLasse Lindahl, et.al, Mapping of ribosomal protein genes by in vitro protein synthesis using DNA fragments of lambda fus3 transducing phage DNA as templates, THE JOURNAL OF BIOLOGICAL CHEMISTRY Vol 252, No 20. Issue of October 25, pp. 7365-7393, 1977.https://pdfs.semanticscholar.org/8d0a/ef8155e29501954fafa585271e3ad3991b25.pdfen_US
dc.identifier.urihttp://hdl.handle.net/11603/14325
dc.language.isoen_USen_US
dc.relation.isAvailableAtThe University of Maryland, Baltimore County (UMBC)
dc.relation.ispartofUMBC Biological Sciences Department Collection
dc.relation.ispartofUMBC Faculty Collection
dc.rightsThis item is likely protected under Title 17 of the U.S. Copyright Law. Unless on a Creative Commons license, for uses protected by Copyright Law, contact the copyright holder or the author.
dc.rightsNon-commercial use only
dc.subjectRibosomal Protein Genesen_US
dc.subjectVitro Protein Synthesisen_US
dc.subjectelectrophoresisen_US
dc.subjecttryptic peptide analysisen_US
dc.titleMapping of ribosomal protein genes by in vitro protein synthesis using DNA fragments of lambda fus3 transducing phage DNA as templatesen_US
dc.title.alternativeMapping of Ribosomal Protein Genes by in Vitro Protein Synthesis Using DNA Fragments of λfus3 Transducing Phage DNA as Templates Phage DNA as Templatesen_US
dc.typeTexten_US

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