Summary
Using electroporation with the phage PRD1 genome, we set up a high-frequency DNA transfer system for a linear dsDNA molecule with 5′-covalently linked terminal proteins. The transfer was saturated when more than 100 ng of PRD1 genome was used. Electroporation efficiency was about four orders of magnitude higher than that obtained with transfection. Removal of the terminal protein abolished plaque formation, which could not be rescued by supplying the terminal protein or phage DNA polymerase or both in trans.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bamford DH, Mindich L (1982) Structure of the lipid-containing bacteriophage PRDI: disruption of wild type and nonsense mutant phage particles with guanidine hydrochloride. J Virol 44:1031–1038
Bamford DH, Rouhiainen L, Takkinen K, Söderlund H (1981) Comparison of the lipid-containing bacteriophages PRDI, PR3, PR4, PR5, L17. J Gen Virol 57:365–373
Bamford DH, Mindich L (1984) Characterization of the DNA-protein complex at the termini of the bacteriophage PRDI genome. J Virol 50:309–315
Bamford DH, McGraw T, Mackenzie G, Mindich L (1983) Identification of a protein covalently bound to the termini of bacteriophage PRD1 DNA. J Virol 47:311–316
Bamford JKH, Bamford DH (1990) Capsomer proteins of bacteriophage PRD1, a bacterial virus with a membrane. Virology 177:445–451
Bamford JKH, Bamford DH (1991) Large scale purification of membrane-containing bacteriophage PRD1 and its subviral particles. Virology 181:348–352
Bamford JKH, Hänninen A-L, Pakula TM, Ojala PM, Kalkbinen N, Frilander M, Bamford DH (1991) Genome organization of a membrane-containing bacteriophage PRDI. Virology 183: (in press)
Bedinger P, Walbot V (1986) DNA synthesis in purified maize mitochondria. Curr Genet 10:631–637
Bradford MM (1976) A rapid sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
Caldentey J, Bamford JKH, Bamford DH (1990) Structure and assembly of bacteriophage PRDI, an Escherichia coli virus with a membrane. J Struct Biol 104:44–51
Carlson JE, Brown GL, Kemble RJ (1986) In organello mitochondrial DNA and RNA synthesis in fertile and cytoplasmic sterile Zea mays L. Curr Genet 11:151–160
Davis TN, Muller ED, Cronan JE Jr (1982) The virion of the lipid-containing bacteriophage PR4. Virology 120:287–306
Dower WJ, Miller JF, Ragsdale CW (1988) High efficiency transformation of E. coli by high voltage electroporation. Nucleic Acids Res 16:6127–6145
Esser K, Kück U, Lang-Hinrichs C, Lemke P, Osiewacz HD, Stahl U, Tudzynski P (1986) Plasmids of eukaryotes. Fundamentals and applications. Springer, Berlin Heidelberg New York Tokyo
Fellay R, Frey J, Krisch H (1987) Interposon mutagenesis of soil and water bacteria: a family of DNA fragments designed for in vitro insertional mutagenesis of gram-negative bacteria. Gene 52:147–154
Hanahan D (1985) Techniques for transformation of E. coli. In: Glover DM (ed) DNA cloning techniques: a practical approach, vol 1. IRL Press, Oxford, pp 109–135
Hirokawa H (1972) Transfecting deoxyribonucleic acid of Bacillus bacteriophage Φ29 that is protease sensitive. Proc Natl Acad Sci USA 69:1555–1559
Hsieh J-C, Yoo S-K, Ito J (1990) An essential arginine residue for initiation of protein-primed DNA replication. Proc Natl Acad Sci USA 87:8665–8669
Kempken F, Meinhardt F, Esser K (1989) In organello replication and viral affinity of linear, extrachromosomal DNA of the ascomycete Ascobolus immersus. Mol Gen Genet 218:523–530
Liljeström P, Laamanen I, Palva ET (1988) Structure and expression of the ompB operon, the regulatory locus for the outer membrane porin regulon in Salmonella typhimurium LT2. J Mol Biol 201:663–673
Mandel M, Higa A (1970) Calcium-dependent bacteriophage DNA infection. J Mol Biol 53:159–162
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY
McGraw T, Yang H-L, Mindich L (1983) Establishment of a physical and genetic map for bacteriophage PRD1. Mol Gen Genet 190:237–244
Mindich L, Bamford DH (1988) Lipid-containing bacteriophages. In: Calendar R (ed) The bacteriophages, vol 2. Plenum, New York, pp 475–520
Mindich L, Cohen J, Weisburd M (1976) Isolation of nonsense suppressor mutants in Pseudomonas. J Bacteriol 126:177–182
Olsen RH, Siak JS, Shipley PL (1974) Characteristics of PRD1, a plasmid dependent broad host range bacteriophage. J Virol 14:698–699
Remaut E, Tsao H, Fiers W (1983) Improved plasmid vectors with a thermoinducible expression and temperature-regulated runaway replication. Gene 22:103–113
Ronda C, López R, Gómez A, García E (1983) Protease-sensitive transfection of Streptococcus pneumoniae with bacteriophage Cp-1 DNA. J Virol 48:721–730
Sakaguchi K (1990) Invertions, a class of structurally and functionally related genetic elements that includes linear DNA plasmids, transposable elements, and genomes of adeno-type viruses. Microbiol Rev 54:66–74
Salas M (1988) Phages with protein attached to the DNA ends. In: Calendar R (ed) The bacteriophages, vol 1. Plenum, New York, pp 169–191
Savilahti H, Caldentey J, Bamford DH (1989) Bacteriophage PRD1 terminal protein: expression of gene VIII in Escherichia coli and purification of the functional P8 product. Gene 85:45–51
Savilahti H, Caldentey J, Lundström K, Syväoja JE, Bamford DH (1991) Overexpression, purification and characterization of Escherichia coli bacteriophage PRD1 DNA polymerase: In vitro synthesis of full-length PRD1 DNA with purified proteins. J Biol Chem (in press)
Sederoff RR, Levings III CS (1985) Supernumerary DNAs in plant mitochondria. In: Hohn B, Dennis ES (eds) Genetic flux in plants. Springer-Verlag, Wien, pp 91–109
Sharp PA, Moore C, Haverty JL (1976) The infectivity of adenovirus 5 DNA-protein complex. Virology 75:442–456
Shigekawa K, Dower WJ (1988) Electroporation of eukaryotes and prokaryotes: a general approach to the introduction of macromolecules into cells. BioTechniques 6:742–751
Tamanoi F (1986) On the mechanism of adenovirus DNA replication. In: Doerfler W (ed) Adenovirus DNA. Martinus Nijhoff, Boston, pp 97–128
Wickner R, Hinnebusch A, Mets L, Lambowitz A, Gunsalus IC, Hollander A (eds) (1987) Extrachromosomal elements in lower eukaryotes. Plenum, New York
Wong FH, Bryan LE (1978) Characteristics of PR5, a lipid-containing plasmid-dependent phage. Can J Microbiol 24:875–882
Author information
Authors and Affiliations
Additional information
Communicated by W. Goebel
Rights and permissions
About this article
Cite this article
Lyra, C., Savilahti, H. & Bamford, D.H. High-frequency transfer of linear DNA containing 5′-covalently linked terminal proteins: electroporation of bacteriophage PRD1 genome into Escherichia coli . Molec. Gen. Genet. 228, 65–69 (1991). https://doi.org/10.1007/BF00282449
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00282449