Nucleic Acids Research, 1986, Vol. 14, No. 24 9679-9698
© 1986
Articles |
Inhibition of restriction endonuclease Nel I cleavage by phosphorothioate groups and its application to oligonucleotide-directed mutagenesis
Max-Planck-Institut für Experimentelle Medizin Abteilung Chemie, Hermann-Rein-Strasse 3, D-3400 Gütingen, FRG
2to whom reprint requests should be addressed
Received October 3, 1986. Accepted November 12, 1986.
M13 RF IV DNA where phosphorothioate groups are incorporated at restriction endonuclease Nci I recognition sites in the (–) strand is efficiently nicked by the action of this enzyme. Incubation of such nicked DNA with exonuclease III produces gapped DNA. The gap can be filled by reaction with deoxynuclease side triphosphates and DNA polymerase I. When this sequence of reactions is performed with DNA containing a mismatch oligonucleotide primer in the (–)strand mutational frequencies of 70–90 % can be obtained upon transformation. The general nature of this methodology has been further shown to be applicable to other restriction enzymes such as Hind II, Pst I and Fsp I. The mutational frequency obtained using these enymes is between 40–80% mainly because of less efficient nicking and gapping. Studies on inhibition of Nci I cleavage show that in addition to a phosphorothioate group at the position of cleavage an additional group in the 5'-neighbouring position is necessary for complete inhibition.
1Present address: Department of Chemistry, Gonzaga University, Spokane, WA 99258, USA
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  T. Hibino Nonfixed Relationship of the Michaelis Constant and Maximum Velocity with Their Corresponding Rate Constants J. Biol. Chem., September 2, 2005; 280(35): 30671 - 30680. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. Truniger, J. M. Lazaro, M. de Vega, L. Blanco, and M. Salas {phi}29 DNA Polymerase Residue Leu384, Highly Conserved in Motif B of Eukaryotic Type DNA Replicases, Is Involved in Nucleotide Insertion Fidelity J. Biol. Chem., August 29, 2003; 278(35): 33482 - 33491. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Motoki, M. Usui, T. Shimazu, M. Hirano, and S. Katoh A Domain of the Manganese-stabilizing Protein from Synechococcus elongatus Involved in Functional Binding to Photosystem II J. Biol. Chem., April 19, 2002; 277(17): 14747 - 14756. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 V. Truniger, J. M. Lazaro, F. J. Esteban, L. Blanco, and M. Salas A positively charged residue of {phi}29 DNA polymerase, highly conserved in DNA polymerases from families A and B, is involved in binding the incoming nucleotide Nucleic Acids Res., April 1, 2002; 30(7): 1483 - 1492. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. D. Drew, M. T. Moss, T. J. Pasieka, C. Grose, W. J. Harris, and A. J. R. Porter Multimeric humanized varicella-zoster virus antibody fragments to gH neutralize virus while monomeric fragments do not J. Gen. Virol., August 1, 2001; 82(8): 1959 - 1963. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. Innes, I. R. Beacham, C.-A. Beven, M. Douglas, M. W. Laird, J. C. Joly, and D. M. Burns The cryptic ushA gene (ushAc) in natural isolates of Salmonella enterica (serotype Typhimurium) has been inactivated by a single missense mutation Microbiology, July 1, 2001; 147(7): 1887 - 1896. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. R. Renzetti, R.-M. Catalioto, M. Criscuoli, P. Cucchi, C. Ferrer, A. Giolitti, M. Guelfi, L. Rotondaro, F. J. Warner, and C. A. Maggi Relevance of Aromatic Residues in Transmembrane Segments V to VII for Binding of Peptide and Nonpeptide Antagonists to the Human Tachykinin NK2 Receptor J. Pharmacol. Exp. Ther., August 1, 1999; 290(2): 487 - 495. [Abstract] [Full Text]
|
|
|
|
 |
|
 H. Shimoi, H. Kitagaki, H. Ohmori, Y. Iimura, and K. Ito Sed1p Is a Major Cell Wall Protein of Saccharomyces cerevisiae in the Stationary Phase and Is Involved in Lytic Enzyme Resistance J. Bacteriol., July 1, 1998; 180(13): 3381 - 3387. [Abstract] [Full Text]
|
|
|
|
|
|
I. Wenderoth, R. Scheibe, and A. von Schaewen Identification of the Cysteine Residues Involved in Redox Modification of Plant Plastidic Glucose-6-phosphate Dehydrogenase J. Biol. Chem., October 24, 1997; 272(43): 26985 - 26990. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Tinti, C. Yang, H. Seo, B. Conti, C. Kim, T. H. Joh, and K.-S. Kim Structure/Function Relationship of the cAMP Response Element in Tyrosine Hydroxylase Gene Transcription J. Biol. Chem., August 1, 1997; 272(31): 19158 - 19164. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 V. Vandevoorde, G. Haegeman, and W. Fiers Induced Expression of Trimerized Intracellular Domains of the Human Tumor Necrosis Factor (TNF) p55 Receptor Elicits TNF Effects J. Cell Biol., June 30, 1997; 137(7): 1627 - 1638. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. K. Han, E. T. Yoon, D. L. Scott, P. B. Sigler, and W. Cho Structural Aspects of Interfacial Adsorption. A CRYSTALLOGRAPHIC AND SITE-DIRECTED MUTAGENESIS STUDY OF THE PHOSPHOLIPASE A2 FROM THE VENOM OF AGKISTRODON PISCIVORUS PISCIVORUS J. Biol. Chem., February 7, 1997; 272(6): 3573 - 3582. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Chen, H. Qi, J. Korenberg, T. A. Garrow, Y.-J. Choi, and B. Shane Purification and Properties of Human Cytosolic Folylpoly-gamma -glutamate Synthetase and Organization, Localization, and Differential Splicing of Its Gene J. Biol. Chem., May 31, 1996; 271(22): 13077 - 13087. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. H. Teckman and D. H. Perlmutter The Endoplasmic Reticulum Degradation Pathway for Mutant Secretory Proteins alpha 1-Antitrypsin Z and S Is Distinct from That for an Unassembled Membrane Protein J. Biol. Chem., May 31, 1996; 271(22): 13215 - 13220. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. A. Keyt, H. V. Nguyen, L. T. Berleau, C. M. Duarte, J. Park, H. Chen, and N. Ferrara Identification of Vascular Endothelial Growth Factor Determinants for Binding KDR and FLT-1 Receptors J. Biol. Chem., March 8, 1996; 271(10): 5638 - 5646. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. N. Fukunaga, M. R. Probst, S. Reisz-Porszasz, and O. Hankinson Identification of Functional Domains of the Aryl Hydrocarbon Receptor J. Biol. Chem., December 8, 1995; 270(49): 29270 - 29278. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. Schmidt, C. Kiecke-Siemsen, A. Waheed, T. Braulke, and K. v. Figura Localization of the Insulin-like Growth Factor II Binding Site to Amino Acids 1508-1566 in Repeat 11 of the Mannose 6-Phosphate/Insulin-like Growth Factor II Receptor J. Biol. Chem., June 23, 1995; 270(25): 14975 - 14982. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M.ía A. Blasco, J. Méndez, J. M. Lázaro, L. Blanco, and M. Salas Primer Terminus Stabilization at the [IMAGE]29 DNA Polymerase Active Site J. Biol. Chem., February 10, 1995; 270(6): 2735 - 2740. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Zhang, L. Chen, V. Villeret, and H. J. Fromm Glycine 122 Is Essential for Cooperativity and Binding of Mg[IMAGE] to Porcine Fructose-1,6-bisphosphatase J. Biol. Chem., January 6, 1995; 270(1): 54 - 58. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Dua, S.-K. Wu, and W. Cho A Structure-Function Study of Bovine Pancreatic Phospholipase A(2) Using Polymerized Mixed Liposomes J. Biol. Chem., January 6, 1995; 270(1): 263 - 268. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L Luo, Y J Liao, L Y Jan, and Y N Jan Distinct morphogenetic functions of similar small GTPases: Drosophila Drac1 is involved in axonal outgrowth and myoblast fusion. Genes & Dev., August 1, 1994; 8(15): 1787 - 1802. [Abstract] [PDF]
|
|
|
|
 |
|
 M. Hatzfeld and M. Burba Function of type I and type II keratin head domains: their role in dimer, tetramer and filament formation J. Cell Sci., July 1, 1994; 107(7): 1959 - 1972. [Abstract] [PDF]
|
|
|
|
 |
|
 T T Nikiforov, R B Rendle, M L Kotewicz, and Y H Rogers The use of phosphorothioate primers and exonuclease hydrolysis for the preparation of single-stranded PCR products and their detection by solid-phase hybridization. Genome Res., April 1, 1994; 3(5): 285 - 291. [Abstract] [PDF]
|
|
|
|
|
|
C. Thiel, M. Osborn, and V. Gerke The tight association of the tyrosine kinase substrate annexin II with the submembranous cytoskeleton depends on intact p11- and Ca(2+)-binding sites J. Cell Sci., November 1, 1992; 103(3): 733 - 742. [Abstract] [PDF]
|
|
|
|
|
|
B Turcotte and L Guarente HAP1 positive control mutants specific for one of two binding sites. Genes & Dev., October 1, 1992; 6(10): 2001 - 2009. [Abstract] [PDF]
|
|
|
|
|
|
S Roberts, T Purton, and D L Bentley A protein-binding site in the c-myc promoter functions as a terminator of RNA polymerase II transcription. Genes & Dev., August 1, 1992; 6(8): 1562 - 1574. [Abstract] [PDF]
|
|
|
|
|
|
S A Emory, P Bouvet, and J G Belasco A 5'-terminal stem-loop structure can stabilize mRNA in Escherichia coli. Genes & Dev., January 1, 1992; 6(1): 135 - 148. [Abstract] [PDF]
|
|
|
|
|
|
X L Liao, L Kretzner, B Seraphin, and M Rosbash Universally conserved and yeast-specific U1 snRNA sequences are important but not essential for U1 snRNP function. Genes & Dev., October 1, 1990; 4(10): 1766 - 1774. [Abstract] [PDF]
|
|
|
|
 |
|
 B. Iverson, S. Iverson, V. Roberts, E. Getzoff, J. Tainer, S. Benkovic, and R. Lerner Metalloantibodies Science, August 10, 1990; 249(4969): 659 - 662. [Abstract] [PDF]
|
|
|
|
|
|
K Zhang, M Noda, W. Vass, A. Papageorge, and D. Lowy Identification of small clusters of divergent amino acids that mediate the opposing effects of ras and Krev-1 Science, July 13, 1990; 249(4965): 162 - 165. [Abstract] [PDF]
|
|
|
|
|
|
E Baldwin and P. Schultz Generation of a catalytic antibody by site-directed mutagenesis Science, September 8, 1989; 245(4922): 1104 - 1107. [Abstract] [PDF]
|
|
|
|
|
|
D L Bentley, W L Brown, and M Groudine Accurate, TATA box-dependent polymerase III transcription from promoters of the c-myc gene in injected Xenopus oocytes. Genes & Dev., August 1, 1989; 3(8): 1179 - 1189. [Abstract] [PDF]
|
|
|
|
|
|
C. Noren, S. Anthony-Cahill, M. Griffith, and P. Schultz A general method for site-specific incorporation of unnatural amino acids into proteins Science, April 14, 1989; 244(4901): 182 - 188. [Abstract] [PDF]
|
|
|
|
|
|
A B Shyu, M E Greenberg, and J G Belasco The c-fos transcript is targeted for rapid decay by two distinct mRNA degradation pathways. Genes & Dev., January 1, 1989; 3(1): 60 - 72. [Abstract] [PDF]
|
|
|
|
|
|
D. Ballard, E Bohnlein, J. Lowenthal, Y Wano, B. Franza, and W. Greene HTLV-I tax induces cellular proteins that activate the kappa B element in the IL-2 receptor alpha gene Science, September 23, 1988; 241(4873): 1652 - 1655. [Abstract] [PDF]
|
|