Summary
A lysine decarboxylase (LDC) gene from Hafnia alvei was cloned in the Escherichia coli strain HB101. A gene bank consisting of 2,000 clones, carrying recombinant plasmids with large DNA fragments of H. alvei integrated in the BamH1 site of pBR322, was screened for LDC activity by a colony filter radioimmunoassay. The gene bank yielded clone 462 expressing high LDC activity with the presence of a plasmid carrying a 7.5 kb insert of H. alvei. Two LDC-positive subclones derived from 462 with inserts of 2.9 and 3.3 kb were sequenced by the shotgun method. An open reading frame for a 83 K protein with 739 amino acids was determined as the coding region for the LDC. The identification of this reading frame as the true reading frame of the H. alvei LDC gene and its similarities with LDC of E. coli are described. The use of the cloned gene for the transformation of plant cells is discussed.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bankier AT, Barrell BG (1983) Shotgun DNA sequencing. Nucleic Acid Biochem B 508:1–34
Barker DG (1982) Cloning and amplified expression of the tyrosyl t-RNA synthetase genes of Bacillus stearothermophilus and Escherichia coli. Eur J Biochem 125:357–360
Birnboim HC, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res 7:1513–1523
Berlin J, Knobloch KH, Höfle G, Witte L (1982) Biochemical characterization of two tobacco cell lines with different levels of cinnamoyl putrescines. J Nat Prod 45:83–88
Berlin J, Beier H, Fecker L, Forche E, Noe W, Sasse F, Schiel O, Wray V (1985) Conventional and new approaches to increase the alkaloid production of plant cell cultures. In: Neuman KH, Barz W, Reinhard E (eds) Primary and secondary metabolism of plant cell cultures. Springer, Heidelberg Berlin New York Tokyo, pp 272–280
Bolivar F (1978) Derivatives of plasmid pBR322 in genetic engineering. In: Boyer HW, Nicosia S (eds) Molecular cloning vectors. Elsevier, North-Holland Biomedical Press, Amsterdam, pp 59–63
Clewell DB (1972) Nature of ColEl plasmid replication in Escherichia coli in the presence of chloramphenicol. J Bacteriol 110:667–676
Dagert M, Ehrlich SD (1979) Prolonged incubation in calcium chloride improves the competence of Escherichia coli cells. Gene 6:23–28
Deus B, Zenk MH (1982) Exploitation of plant cells for the production of natural compounds. Biotechnol Bioeng 24:1965–1974
Di Cosmo F (1984) Stress and secondary metabolism in cultured plant cells. In: Timmermann BN, Steelink C, Loewus FA (eds) Phytochemical adaptations to stress. Plenum, New York, pp 97–175
Fecker L, Braun V (1983) Cloning and expression of the fhu genes involved in iron (III)-hydroxamate uptake by Escherichia coli. J Bacteriol 156:1301–1314
Horejsci V, Hilgert I (1983) Nitrocellulose membrane as an antigen or antibody carrier for screening hybridoma cultures. J Immunol Methods 62:325–329
Ikemura T (1981) Correlation between the abundance of Escherichia coli transfer RNAs and the occurrence of the respective codons in its protein genes. J Mol Biol 146:1–21
Kemp DJ, Cowman AF (1981) Direct immunoassay for detecting Escherichia coli colonies that contain polypeptides encoded by cloned DNA segments. Proc Natl Acad Sci USA 78:4520–4524
Maniatis T, Fritsch EE, Sambrock J (1982) Molecular cloning. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
Maxam AM, Gibert W (1980) Sequencing end-labeled DNA with base-specific chemical cleavage. Methods Enzymol 65:499–560
Messing J (1983) New M13 Vectors for cloning. Methods Enzymol 101:20–78
Phan APH, Ngo TT, Lehnhoff HM (1982) Spectrometric assay for lysine decarboxylase. Anal Biochem 120:193–197
Rosenberg M, Court D (1979) Regulatory sequences involved in the promotion and termination of RNA transcription. Annu Rev Genet 13:319–351
Sabo DL, Boeker EA, Byers B, Waron H, Fischer EH (1974) Purification and physical properties of inducible Escherichia coli lysine decarboxylase. Biochemistry 13:662–669
Sabo DL, Fischer EH (1974) Chemical properties of Escherichia coli lysine decarboxylase including a segment of its pyridoxal-5′-phosphate binding site. Biochemistry 13:670–676
Sakazaki R (1981) The genus Hafnia. In: Starr MP, Stolp H, Trüper HG, Balows A, Schlegel HG (eds) The prokaryotes. Springer, Berlin Heidelberg New York, pp 1181–1186
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
Sasse F, Knobloch KH, Berlin J (1982) Induction of secondary metabolism in cell suspension cultures of Catharanthus roseus, Nicotiana tabacum and Peganum harmala. In: Fjiwara A (ed) Plant tissue culture 1982 Maruzen Co, Tokyo, pp 343–344
Schoofs G, Teichmann S, Hartmann T, Wink M (1983) Lysine decarboxylase in plants and its integration in quinolizidine alkaloid biosynthesis. Phytochemistry 22:65–69
Shine J, Dalgarno L (1974) The 3′-terminal sequence of Escherichia coli 16 S ribosomal RNA: complementarity to nonsense triplets and ribosome binding sites. Proc Natl Acad Sci USA 71:1342–1346
Staden R (1977) Sequence data handling by computer. Nucleic Acids Res 4:4037–4053
Steitz JA, Jakes K (1975) How ribosomes select initiator regions in m-RNA: Base pair formation between the 3′ terminus of 16 S rRNA and the mRNA during initiation of protein synthesis in E. coli. Proc Natl Acad Sci USA 72:4734–4738
Stüber D, Bujard H (1981) Organization of transcriptional signals in plasmid pBR322 and pACYC184. Proc Natl Acad Sci USA 78:167–171
Sutcliffe JG (1978) Complete nucleotide sequence of the Escherichia coli plasmid pBR322. Cold Spring Harbor Symp Quant Biol 43: 77–90
Tabor H, Hafner EW, Tabor CW (1980) Construction of an Escherichia coli strain unable to synthesize putrescine, spermidine or cadaverine: characterization of two genes controlling lysine decarboxylase. J Bacteriol 144:952–956
Timmis KN, Cabello F, Cohen SN (1978) Cloning and characterization of EcoR1 and HindIII restriction and endonucleasegenerated fragments of antibiotic resistance plasmids R6-5 and R6. Mol Gen Genet 162:121–137
Wink M, Hartmann T (1982) Localization of quinolizidine alkaloid biosynthesis in leaf chloroplasts of Lypinus polyphyllus. Plant Physiol 70:74–77
Wink M, Hartmann T, Schiebel HM (1979) A model mechanism for enzymatic synthesis of lupin alkaloids. Z Naturforsch 35c:704–709
Wink M, Hartmann T, Witte L (1980) Biotransformation of cadaverine and potential biogenetic intermediates of lupanine biosynthesis by plant cell suspension cultures. Planta Medica 40:31–39
Author information
Authors and Affiliations
Additional information
Communicated by J. Schell
Rights and permissions
About this article
Cite this article
Fecker, L.F., Beier, H. & Berlin, J. Cloning and characterization of a lysine decarboxylase gene from Hafnia alvei . Mol Gen Genet 203, 177–184 (1986). https://doi.org/10.1007/BF00330400
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00330400