Abstract
A recombinant clone, pM52, containing cDNA for maize phosphoenolpyruvate carboxylase (PEPCase, EC 4.1.1.31) was isolated from a maize leaf cDNA library constructed using an expression vector in Escherichia coli. The screening of the clone was conveniently performed through its ability to complement the phenotype (glutamate requirement) of PEPCase-negative mutant of E. coli. The enzyme encoded by this clone was identical with the major PEPCase in maize, a key enzyme in the C4-pathway, as judged from its allosteric properties and immunological reactivity. The cloned cDNA (3093 nucleotides in length) contained an open reading frame of 2805 nucleotides, the 3'-untranslated region of 222 nucleotides and the poly(dA) tract of 64 nucleotides. The deduced amino acid sequence (935 residues) of the enzyme showed higher homology with that of an enterobacterium, E. coli (43%) than that of a cyanobacterium (blue-green alga), Anacystis nidulans (33%).
Full text
PDF













Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Ashworth J. M., Kornberg H. L. The anaplerotic fixation of carbon dioxide by Escherichia coli. Proc R Soc Lond B Biol Sci. 1966 Aug 16;165(999):179–188. doi: 10.1098/rspb.1966.0063. [DOI] [PubMed] [Google Scholar]
- Aviv H., Leder P. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408–1412. doi: 10.1073/pnas.69.6.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Davidson J. N., Niswander L. A. Partial cDNA sequence to a hamster gene corrects defect in Escherichia coli pyrB mutant. Proc Natl Acad Sci U S A. 1983 Nov;80(22):6897–6901. doi: 10.1073/pnas.80.22.6897. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Deininger P. L. Random subcloning of sonicated DNA: application to shotgun DNA sequence analysis. Anal Biochem. 1983 Feb 15;129(1):216–223. doi: 10.1016/0003-2697(83)90072-6. [DOI] [PubMed] [Google Scholar]
- Dennis E. S., Gerlach W. L., Pryor A. J., Bennetzen J. L., Inglis A., Llewellyn D., Sachs M. M., Ferl R. J., Peacock W. J. Molecular analysis of the alcohol dehydrogenase (Adh1) gene of maize. Nucleic Acids Res. 1984 May 11;12(9):3983–4000. doi: 10.1093/nar/12.9.3983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fujita N., Miwa T., Ishijima S., Izui K., Katsuki H. The primary structure of phosphoenolpyruvate carboxylase of Escherichia coli. Nucleotide sequence of the ppc gene and deduced amino acid sequence. J Biochem. 1984 Apr;95(4):909–916. doi: 10.1093/oxfordjournals.jbchem.a134718. [DOI] [PubMed] [Google Scholar]
- Hanahan D. Studies on transformation of Escherichia coli with plasmids. J Mol Biol. 1983 Jun 5;166(4):557–580. doi: 10.1016/s0022-2836(83)80284-8. [DOI] [PubMed] [Google Scholar]
- Izui K., Sabe H., Katsuki H. Increased synthesis of phosphoenolypyruvate carboxylase in a strain of Escherichia coli bearing a ColE1-ppc+ hybrid plasmid. FEBS Lett. 1981 Oct 26;133(2):311–315. doi: 10.1016/0014-5793(81)80531-5. [DOI] [PubMed] [Google Scholar]
- Izui K., Taguchi M., Morikawa M., Katsuki H. Regulation of Escherichia coli phosphoenolpyruvate carboxylase by multiple effectors in vivo. II. Kinetic studies with a reaction system containing physiological concentrations of ligands. J Biochem. 1981 Nov;90(5):1321–1331. doi: 10.1093/oxfordjournals.jbchem.a133597. [DOI] [PubMed] [Google Scholar]
- Kodaki T., Katagiri F., Asano M., Izui K., Katsuki H. Cloning of phosphoenolpyruvate carboxylase gene from a cyanobacterium, Anacystis nidulans, in Escherichia coli. J Biochem. 1985 Feb;97(2):533–539. doi: 10.1093/oxfordjournals.jbchem.a135088. [DOI] [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Lonberg N., Gilbert W. Intron/exon structure of the chicken pyruvate kinase gene. Cell. 1985 Jan;40(1):81–90. doi: 10.1016/0092-8674(85)90311-3. [DOI] [PubMed] [Google Scholar]
- Messing J. New M13 vectors for cloning. Methods Enzymol. 1983;101:20–78. doi: 10.1016/0076-6879(83)01005-8. [DOI] [PubMed] [Google Scholar]
- Norrander J., Kempe T., Messing J. Construction of improved M13 vectors using oligodeoxynucleotide-directed mutagenesis. Gene. 1983 Dec;26(1):101–106. doi: 10.1016/0378-1119(83)90040-9. [DOI] [PubMed] [Google Scholar]
- Okayama H., Berg P. High-efficiency cloning of full-length cDNA. Mol Cell Biol. 1982 Feb;2(2):161–170. doi: 10.1128/mcb.2.2.161. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sabe H., Miwa T., Kodaki T., Izui K., Hiraga S., Katsuki H. Molecular cloning of the phosphoenolpyruvate carboxylase gene, ppc, of Escherichia coli. Gene. 1984 Nov;31(1-3):279–283. doi: 10.1016/0378-1119(84)90222-1. [DOI] [PubMed] [Google Scholar]
- Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Scrutton M. C., Fatebene F. An assay system for localisation of pyruvate and phosphoenolpyruvate carboxylase activity on polyacrylamide gels and its application to detection of these enzymes in tissue and cell extracts. Anal Biochem. 1975 Nov;69(1):247–260. doi: 10.1016/0003-2697(75)90584-9. [DOI] [PubMed] [Google Scholar]
- Sims T. L., Hague D. R. Light-stimulated increase of translatable mRNA for phosphoenolpyruvate carboxylase in leaves of maize. J Biol Chem. 1981 Aug 25;256(16):8252–8255. [PubMed] [Google Scholar]
- Sugiyama T., Mizuno M., Hayashi M. Partitioning of Nitrogen among Ribulose-1,5-bisphosphate Carboxylase/Oxygenase, Phosphoenolpyruvate Carboxylase, and Pyruvate Orthophosphate Dikinase as Related to Biomass Productivity in Maize Seedlings. Plant Physiol. 1984 Jul;75(3):665–669. doi: 10.1104/pp.75.3.665. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sutcliffe J. G. Complete nucleotide sequence of the Escherichia coli plasmid pBR322. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 1):77–90. doi: 10.1101/sqb.1979.043.01.013. [DOI] [PubMed] [Google Scholar]
- Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Uedan K., Sugiyama T. Purification and characterization of phosphoenolpyruvate carboxylase from maize leaves. Plant Physiol. 1976 Jun;57(6):906–910. doi: 10.1104/pp.57.6.906. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yoshinaga T., Teraoka H., Izui K., Katsuki H. Molecular properties of phosphoenolpyruvate carboxylase of Escherichia coli W. J Biochem. 1974 Apr;75(4):913–924. doi: 10.1093/oxfordjournals.jbchem.a130465. [DOI] [PubMed] [Google Scholar]