Home LiteratureArticle Details
PMID: 6276682 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Cloning of E. coli pnp gene from an episome.

Molecular & general genetics : MGG ·Vol. 183 ·No. 2 ·1981-00-00 ·Pages 298-305

Portier C, Migot C, Grumberg-Manago M

Abstract

Starting with an F' episome harboring a transposon inserted in the pnp gene (Portier 1980), we were able to identify an EcoRI restriction fragment carrying the pnp and argG genes. This fragment, from both wild-type and mutant episomes, was cloned ni pACYC184. The presence of argG on the fragment allowed positive selection of the desired clones in an auxotrophic strain (argG). A restriction map was established and a fragment of 3 megadaltons subcloned in the plasmid vector pBR322. The pnp gene corresponds to about 50% of this subcloned segment and was roughly located by deletion mapping. The direction of transcription and locations of the promotor and gene extremities were determined by analyzing proteins synthesized in "maxi-cells". In addition, the gene coding for a 10,000 dalton protein was found to reside adjacent to the beginning of the pnp structural gene. Strains carrying plasmids which express the pnp overproduce polynucleotide phosphorylase.

MeSH Terms
Arginine/genetics Bacterial Proteins/genetics Chromosome Mapping DNA Restriction Enzymes/metabolism DNA, Recombinant/analysis Escherichia coli/genetics Genes Genes, Bacterial Genetic Linkage Plasmids
Chemicals
Bacterial Proteins DNA, Recombinant Arginine DNA Restriction Enzymes
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Portier C
Migot C
Grumberg-Manago M
References (32)
32 references, click to expand
  1. The proteolytic conversion of polynucleotide phosphorylase to a primer-dependent form.
    J Biol Chem. 1969 May 25;244(10):2558-66 PMID: 5769990
  2. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  3. Beta-galactosidase. Rates of synthesis and degradation of incomplete chains.
    J Biol Chem. 1972 Apr 10;247(7):2205-11 PMID: 4552691
  4. Primary structure of the 16S rRNA binding protein S15 from Escherichia coli ribosomes.
    FEBS Lett. 1976 May 1;64(2):307-9 PMID: 776686
  5. Attenuation and processing of RNA from the rplJL--rpoBC transcription unit of Escherichia coli.
    Proc Natl Acad Sci U S A. 1980 Jun;77(6):3331-5 PMID: 6158044
  6. Degradation of Escherichia coli polynucleotide phosphorylase by E. coli endogenous proteases and by trypsin.
    Biochimie. 1971;53(3):291-302 PMID: 4935403
  7. DISC ELECTROPHORESIS. II. METHOD AND APPLICATION TO HUMAN SERUM PROTEINS.
    Ann N Y Acad Sci. 1964 Dec 28;121:404-27 PMID: 14240539
  8. Quaternary structure of polynucleotide phosphorylase from Escherichia coli: evidence of a complex between two types of polypeptide chains.
    Eur J Biochem. 1975 Jul 15;55(3):573-82 PMID: 1100386
  9. Plasmid identification using specific endonucleases.
    Mol Gen Genet. 1974;133(2):141-9 PMID: 4614064
  10. Nonchromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R-factor DNA.
    Proc Natl Acad Sci U S A. 1972 Aug;69(8):2110-4 PMID: 4559594
  11. [Separation and identification of polynucleotide phosphorylase by electrophoresis on polyacrylamide gel].
    C R Acad Sci Hebd Seances Acad Sci D. 1967 Dec 6;265(23):1823-6 PMID: 4967519
  12. Functional mRNA half lives in E. coli.
    Mol Gen Genet. 1978 Nov 9;166(3):329-36 PMID: 368581
  13. Genes coding for ribosomal proteins S15, L21, and L27 map near argG in Escherichia coli.
    J Bacteriol. 1979 Jun;138(3):832-8 PMID: 378941
  14. Synthesis and degradation of lac mRNA in E. coli depleted of 30S ribosomal subunits.
    Mol Gen Genet. 1979 Jun 7;173(2):135-44 PMID: 386032
  15. The control region of the F sex factor DNA transfer cistrons: physical mapping by deletion analysis.
    Mol Gen Genet. 1979 Jan 16;169(1):49-57 PMID: 374998
  16. Simple method for identification of plasmid-coded proteins.
    J Bacteriol. 1979 Jan;137(1):692-3 PMID: 368040
  17. Characterization of polynucleotide phosphorylase mutants of Escherichia coli.
    J Bacteriol. 1969 Mar;97(3):1437-43 PMID: 4887520
  18. Construction of a physical map of a kanamycin (Km) transposon, Tn5, and a comparison to another Km transposon, Tn903.
    Mol Gen Genet. 1980 Apr;178(1):77-83 PMID: 6247620
  19. A rapid alkaline extraction procedure for screening recombinant plasmid DNA.
    Nucleic Acids Res. 1979 Nov 24;7(6):1513-23 PMID: 388356
  20. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system.
    Gene. 1977;2(2):95-113 PMID: 344137
  21. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid.
    J Bacteriol. 1978 Jun;134(3):1141-56 PMID: 149110
  22. Further characterization of the polynucleotide phosphorylase of Micrococcus luteus.
    Nucleic Acids Res. 1975 Feb;2(2):149-64 PMID: 1121422
  23. Complete nucleotide sequence of the Escherichia coli plasmid pBR322.
    Cold Spring Harb Symp Quant Biol. 1979;43 Pt 1:77-90 PMID: 383387
  24. A restriction enzyme cleavage map of Tn5 and location of a region encoding neomycin resistance.
    Mol Gen Genet. 1979;177(1):65-72 PMID: 231729
  25. Recalibrated linkage map of Escherichia coli K-12.
    Bacteriol Rev. 1976 Mar;40(1):116-67 PMID: 773363
  26. Biochemical characterization of nonintegrated plasmid-folded chromosome complexes: sex factor F and the Escherichia coli nucleoid.
    J Bacteriol. 1979 Feb;137(2):885-90 PMID: 370105
  27. Study on the structure-function relationship of polynucleotide phosphorylase: model of a proteolytic degraded polynucleotide phosphorylase.
    Nucleic Acids Res. 1976 Nov;3(11):3015-24 PMID: 794831
  28. Isolation of a polynucleotide phosphorylase mutant using a kanamycin resistant determinant.
    Mol Gen Genet. 1980;178(2):343-9 PMID: 6248726
  29. Increased expression of a eukaryotic gene in Escherichia coli through stabilization of its messenger RNA.
    Proc Natl Acad Sci U S A. 1979 Nov;76(11):5774-8 PMID: 160556
  30. Isolation and mapping of polynucleotide phosphorylase mutants of Escherichia coli.
    J Bacteriol. 1969 Mar;97(3):1431-6 PMID: 4887519
  31. Expression of the HIS3 gene of Saccharomyces cerevisiae in polynucleotide phosphorylase-deficient strains of Escherichia coli K-12.
    Gene. 1980 Dec;12(1-2):1-10 PMID: 7011901
  32. Genetic studies of the ribosomal proteins in Escherichia coli. XI. Mapping of the genes for L21, L27, S15 and S21 by using hybrid bacteria and over-production of these proteins in the merodiploid strains.
    Mol Gen Genet. 1978 Apr 6;160(2):151-5 PMID: 349351
Article Info
Journal
Molecular & general genetics : MGG
Abbr.
Mol Gen Genet
ISSN
0026-8925
DOI
10.1007/BF00270632
Published
1981-00-00
Pages
298-305
Language
English
Region
Germany
NLM ID
0125036
Subset
IM
External Links
PubMed source DOI: 10.1007/BF00270632
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2