Isolation of transformation-deficient Streptococcus pneumoniae mutants defective in control of competence, using insertion-duplication mutagenesis with the erythromycin resistance determinant of pAM beta 1
Journal of BacteriologyPublished 1 September 1984Open access
Donald A. Morrison, Marie‐Claude Trombe, Melody Hayden, Gregory A. Waszak, J D Chen
Citations108
SJR quartileQ2
SJR score1.09
SNIP0.85
Generate an AI Snapshot to get a quick, structured summary of this paper.
Study Snapshot
ObjectiveStudy objective
MethodsResearch methodology
PopulationPopulation studied
Sample sizeSample sizes
OutcomesStudy outcomes here
ResultsStudy results comes here
LimitationsResearch study limitations comes here
A concise AI-generated summary of the paper will appear here once you click Generate AI Snapshot.
TL;DR
Several transformation-deficient mutants of Streptococcus pneumoniae were isolated after insertion-duplication mutagenesis and characterized as defective in the control of competence induction, possibly due to a block in the production of the intercellular competence-inducing protein.
Abstract
Several transformation-deficient mutants of Streptococcus pneumoniae were isolated after insertion-duplication mutagenesis. Mutagenesis was accomplished by transformation of competent cells with chimeric DNA formed by the ligation of TaqI fragments of pneumococcal DNA to the erythromycin resistance determinant of the streptococcal plasmid pAM beta 1. The two mutants described were characterized as defective in the control of competence induction, possibly due to a block in the production of the intercellular competence-inducing protein.
Keywords
MedicineBiochemistry, Genetics and Molecular Biology
European Journal of BiochemistryQuantitative Film Detection of 3H and 14C in Polyacrylamide Gels by Fluorography
4,687 Citations1975Ronald A. Laskey, Anthony D. Mills
It is demonstrated that pre-exposure of the film to a brief flash of light greatly increases the sensitivity of fluorography, because it corrects the non-linear relationship between radioactivity of the sample and absorbance of theFilm image.
Proceedings of the National Academy of SciencesTransformation of yeast.
2,226 Citations1978A. Hinnen, James Hicks +1 more
This work has used recently developed hybridization and restriction endonuclease mapping techniques to demonstrate directly the presence of the transforming DNA in the yeast genome and also to determine the arrangement of the sequences that were introduced.
Journal of BacteriologyCharacterization of Three Plasmid Deoxyribonucleic Acid Molecules in a Strain of <i>Streptococcus faecalis</i> : Identification of a Plasmid Determining Erythromycin Resistance
325 Citations1974Don B. Clewell, Yusuke Yagi +2 more
Three plasmids designated alpha, beta, and gamma, distinguishable by their molecular weights, were isolated from Streptococcus faecalis strain DS-5 (ATCC 14508).
ScienceLethal Disruption of the Yeast Actin Gene by Integrative DNA Transformation
287 Citations1982David Shortle, James E. Haber +1 more
A mutant allele of the chromosomal locus corresponding to the cloned actin gene of the yeast Saccharomyces cerevisiae has been constructed by DNA transformation with a hybrid plasmid which integrates into, and thereby disrupts, the protein-encoding sequences of the gene.
Journal of BacteriologyConstruction and properties of an integrable plasmid for Bacillus subtilis
263 Citations1983Franco Ferrari, Anh Hoang Nguyen +2 more
The results suggest that production of the tetracycline gene product is lethal to B. subtilis.
Analytical BiochemistryA procedure for the large-scale isolation of highly purified plasmid DNA using alkaline extraction and binding to glass powder
261 Citations1982Močibob Marko, R. Chipperfield +1 more
In the present method, all detectable RNA, chromosomal DNA, and protein are removed without the use of enzymes, phenol extraction, dialysis, or equilibrium centrifugation.
Proceedings of the National Academy of SciencesREGULATION OF THE TRANSFORMABILITY OF PNEUMOCOCCAL CULTURES BY MACROMOLECULAR CELL PRODUCTS
250 Citations1964Alexander Tomasz, Rollin D. Hotchkiss
Journal of General MicrobiologyOn the Nature of Competence of Transformable Streptococci
205 Citations1963R Pakuła, W. Walczak
Transformable streptococci produce an exocellular factor provoking competence under certain conditions, and non-competent cultures become competent upon addition of this factor.
Journal of BacteriologyMutants of <i>Diplococcus pneumoniae</i> that Lack Deoxyribonucleases and Other Activities Possibly Pertinent to Genetic Transformation
187 Citations1970Sanford A. Lacks
Mutants of Diplococcus pneumoniae that lacked the two major deoxyribonucleases of the cell were obtained, indicating that a particular cellular component that is susceptible to loss by mutation, such as an enzyme, is responsible for low integration efficiency.
GeneStable integration of foreign DNA into the chromosome of the cyanobacterium Synechococcus R2
166 Citations1983John G. Williams, Aladar A. Szalay
Transformation in Synechococcus R2 can be exploited to modify precisely and extensively the genome of this photosynthetic microorganism.
Proceedings of the National Academy of SciencesOn the nature of the pneumococcal activator substance.
127 Citations1966Alexander Tomasz, Jean Mosser
Procedures for the growth of pneumococcal cultures in the competent and incompe- tent states and for assaying the viable titer of cultures have been described and a low pH-C medium was introduced for the routine growth of incompetent cultures.
Molecular and General Genetics MGGDestruction of low efficiency markers is a slow process occurring at a heteroduplex stage of transformation
124 Citations1974Nadja B. Shoemaker, Walter R. Guild
Assays of the transforming activity of donor markers in lysates made after various times of incubation of recipient cells following exposure to DNA imply that about as many of the surviving low efficiency markers have simply escaped attack as have been inserted into both strands by the excision-repair process suggested by Ephrussi-Taylor.
Journal of BacteriologyModel for the Mechanism Controlling the Expression of Competent State in Pneumococcus Cultures
112 Citations1966Alexander Tomasz
A physiological model is proposed for the control of the competent state in pneumococcus and evidence is presented for physiological influences affecting the cell's capacity to react with the activator.
NatureCompetence for genetic transformation in pneumococcus depends on synthesis of a small set of proteins
111 Citations1979Donald A. Morrison, Maggie Baker
Pulse-labelling studies show that for the brief period of competence protein synthesis is restricted to a few specific polypeptides, and the response to competence factor has been shown to require protein synthesis.
Proceedings of the National Academy of SciencesMechanism of integrating foreign DNA during transformation of Bacillus subtilis.
105 Citations1978Craig H. Duncan, G A Wilson +1 more
Results indicated that the entire chimeric plasmid can be incorporated into the chromosome of B. subtilis by a Campbell-like model, and an additional mechanism for transformation exists whereby plasmids can be integrated if sufficient chromosomal homology is maintained.
Journal of BacteriologyIsolation and characterization of three new classes of transformation-deficient mutants of Streptococcus pneumoniae that are defective in DNA transport and genetic recombination
96 Citations1983Donald A. Morrison, Sanford A. Lacks +2 more
Transformation-deficient mutants of Streptococcus pneumoniae were isolated after nitrosoguanidine mutagenesis and developed normal peaks of competence, as tested by their ability to degrade one strand of donor DNA, but they yielded transformants for chromosomal point markers at efficiencies below that of the wild type.
Journal of Molecular BiologyFate of donor DNA in pneumococcal transformation
94 Citations1967Sanford A. Lacks, Bill Greenberg +1 more
It is concluded that single strands are precursors of genetically integrated DNA and that recombination involves the interaction of homologous single strands with host DNA.
The Journal of General PhysiologyGenetic Studies of Recombining DNA in Pneumococcal Transformation
88 Citations1966Harriett Ephrussi-Taylor, Thomas C. Gray
It is proposed, on the basis of genetic evidence, that following essentially random exchanges between donor DNA and recipient chromosome, a revision process occurs, specific for certain types of mutated sites, which appears to remove preferentially donor DNA sequences from the primary recombinant structure, and allow repair along the chromosomal template, leading to low efficiency in the genetic integration of these sites.
GeneRapid cloning of specific DNA fragments of Streptococcus pneumoniae by vector integration into the chromosome followed by endonucleolytic excision
79 Citations1981Vincent Méjean, Jean‐Pierre Claverys +2 more
Host sequences adjacent to the site of insertion are easily cloned by enzymatic excision and recircularization of the plasmid, followed by propagation in Escherichia coli, particularly useful for repeated cloning of a given fragment that carries various mutations.
Journal of Molecular BiologyPhysical and genetic hybrids formed in bacterial transformation
74 Citations1968Theodore Gurney, Maurice S. Fox
It is concluded that DNA can be integrated into the recipient genome in a continuum of lengths ranging from an undetermined lower limit below 1 × 106 daltons to more than 5 × 106Daltons, in single-stranded molecular weight.
Journal of BacteriologyCompetence for Deoxyribonucleic Acid Uptake and Deoxyribonuclease Action External to Cells in the Genetic Transformation of <i>Diplococcus pneumoniae</i>
71 Citations1973Sanford A. Lacks, Bill Greenberg
A mutant of Diplococcus pneumoniae that apparently does not require activator can become competent for uptake of deoxyribonucleic acid (DNA) when grown in dilute cultures or in the presence of trypsin, but development of competence in both mutant and wild strains is temperature dependent.
Biochimica et Biophysica Acta (BBA) - Nucleic Acids and Protein SynthesisBreakage prior to entry of donor DNA in Pneumococcus transformation
71 Citations1973Donald A. Morrison, Walter R. Guild
The results suggest that double-strand breaks precede entry, that the strand segments thus produced enter without further breakage, and that the complement of each entering segment probably is degraded with release of the products outside the cell.
Cold Spring Harbor Symposia on Quantitative Biology
68 Citations1946
This paper presented by McCarty and Harriet Taylor at the June 1946 Cold Spring Harbor Symposium was part of his effort to bring his work on transformation to "some kind of reasonable conclusion" prior to his leaving the lab that July.
PlasmidA new technique for genetic engineering of Agrobacterium Ti plasmid
68 Citations1983Luca Comai, Christine Schilling-Cordaro +2 more
A new technique is described that allows easy introduction of foreign genetic elements into specific regions of Agrobacterium tumefaciens DNA through the use of plasmids that have a region homologous to the genome of the recipient.
Journal of BacteriologyTransformation and Deoxyribonucleic Acid Size: Extent of Degradation on Entry Varies with Size of Donor
65 Citations1972Donald A. Morrison, Walter R. Guild
The probability that a marker in the intact fraction will be integrated, as a function of the length of the donor strand after entry, is calculated, which has a linear dependence on strand length for activities below 40% of maximum, and extrapolates to zero activity at 77,000 daltons per strand.
Binding and Entry of DNA in Bacterial Transformation
65 Citations1977Sanford A. Lacks
Molecular and General Genetics MGGExcision and repair of mismatched base pairs in transformation of Streptococcus pneumoniae
51 Citations1980J.P. Claverys, M. Roger +1 more
The transfer of information from each donor DNA strand to the recipient genome is studied, on the basis of differences in the rates of phenotypic expression of a given marker introduced on opposite strands, to strongly suggest that the hex function is an excision-repair system acting on donor-recipient base pair mismatches.
GeneAmplification of a chimeric plasmid carrying an erythromycin-resistance determinant introduced into the genome of Streptococcus pneumoniae
42 Citations1983Homa Vasseghi, Jean‐Pierre Claverys
Results indicate that, in the orientation conferring a high-level resistance without amplification, the ery region is transcribed both from its own promoter and from the promoter of the amiA locus.
Journal of BacteriologyPurification and Properties of Streptococcal Competence Factor Isolated from Chemically Defined Medium
37 Citations1972C. Gomez Leonard, Roger M. Cole
The data indicate that competence factor is a small, dialyzable, highly basic compound that is free from lipids, phosphorus, and carbohydrates, and is colorless and thermoresistant.
GeneCloning of Streptococcus pneumoniae DNA: Its use in pneumococcal transformation and in studies of mismatch repair
29 Citations1981J.P. Claverys, J M Louarn +1 more
EcoRI fragments of the amiA locus in Streptococcus pneumoniae were cloned either into a derivative of lambda or into pBR325 plasmid, which confer resistance to aminopterin.
Journal of BacteriologyStructure of Deoxyribonucleic Acid on the Cell Surface During Uptake by Pneumococcus
28 Citations1973Donald A. Morrison, Walter R. Guild
It was found that entry past the cell barrier to deoxyribonuclease could occur at 0 C by a process sensitive to ethylenediaminetetraacetate, implying that the breaks produced outside the cell are not at genetically specific sites.
Proceedings of the National Academy of SciencesSwitches in macromolecular synthesis during induction of competence for transformation of Streptococcus sanguis.
23 Citations1980Jawahar L. Raina, Arnold W. Ravin
One of the newly synthesized early polypeptides, E16, is shown to be a component of the presynaptic complex containing single-stranded DNA that is produced in vivo upon uptake of native donor DNA.
Molecular and Cellular BiologyEfficient Transformation of <i>Dictyostelium discoideum</i> Amoebae
16 Citations1983Stephen L. Barclay, Erna Meller
The results suggest that direct inactivation and recovery of genes that are essential for differentiation of D. discoideum will be possible and that integration can result in insertional inactivation of genes important for differentiation but not for growth.
GeneticsINTEGRATION EFFICIENCY IN DNA-INDUCED TRANSFORMATION OF PNEUMOCOCCUS. I. A METHOD OF TRANSFORMATION IN SOLID MEDIUM AND ITS USE FOR ISOLATION OF TRANSFORMATION-DEFICIENT AND RECOMBINATION-MODIFIED MUTANTS
15 Citations1973Gérard Tiraby, Jean‐Pierre Claverys +1 more
Most of the mutants found are transformation-defective and show a large diversity in their response to ultraviolet light, and some of these mutants have lost their ability to take up transforming DNA.
Molecular and Cellular BiologyEfficient transformation of Dictyostelium discoideum amoebae.
10 Citations1983Stephen L. Barclay, E Meller
BIOCHEMICAL STUDIES OF ENVIRONMENTAL FACTORS ESSENTIAL IN TRANSFORMATION OF PNEUMOCOCCAL TYPES
3 Citations1946Maclyn Mccarty, Harriett E. Taylor +1 more