scholarly journals GENETIC HETEROZYGOSITY IN UNREPLICATED BACTERIOPHAGE Λ RECOMBINANTS

Genetics ◽  
1975 ◽  
Vol 81 (1) ◽  
pp. 33-50
Author(s):  
Raymond L White ◽  
Maurice S Fox
Keyword(s):  
Dna Synthesis ◽  
Base Pair ◽  
Recombinant Dna ◽  
Dna Molecules ◽  
Bacteriophage Λ ◽  
Negative Interference ◽  
Genetic Interval ◽  
Parental Material ◽  
Genetic Heterozygosity ◽  
Genetic Contributions

ABSTRACT Bacteriophage crosses using density-labeled parents have been carried out under conditions restricting DNA synthesis. The parental material and genetic contributions to progeny manifesting recombination within a genetic interval sufficiently short to exhibit high negative interference have been examined. The unreplicated products of recombination isolated as phage particles appear to contain long continuous heteroduplex regions which are heterozygous for the closely linked markers. Recombination between closely linked markers seems to be the consequence of the removal of base-pair mismatches that are present within the heteroduplex regions. This localized reduction of heterozygosity within the heteroduplex regions that join the parental components of recombinant DNA molecules can account for high negative interference.

Rare internal C4A4 repeats in the micronuclear genome of Oxytricha fallax

1984 ◽  
Vol 4 (12) ◽  
pp. 2661-2667
Author(s):  
D Dawson ◽  
G Herrick
Keyword(s):  
Base Pair ◽  
Full Length ◽  
Dna Molecules ◽  
Base Pairs ◽  
Macronuclear Development ◽  
Structural Relationships

Approximately 20,000 different short, linear, macronuclear DNA molecules are derived from micronuclear sequences of Oxytricha fallax after conjugation. These macronuclear DNAs are terminated at both ends by 20 base pairs of the sequence 5'-dC4A4-3'. Sequences homologous to this repeat (C4A4+) are also abundant in the micronuclear chromosomes, but most reside at their telomeres. Here we show that nontelomeric C4A4 clusters of 20 base pairs or longer exist in only a few hundred copies per micronuclear genome. This demonstrates that nearly none of the 20,000 sequence blocks of micronuclear DNA destined to be macronuclear DNA molecules can be flanked by full-length (20-base pair) C4A4 clusters, and therefore C4A4 repeats must be added to most, if not all, macronuclear telomeres during macronuclear development. Six internal micronuclear C4A4+ loci were cloned, and their structural relationships with macronuclear and micronuclear sequences were examined. The possible origins and functions of these rare, micronuclear internal C4A4 loci are discussed.

Tolerance of Base Pair Size and Shape in Postlesion DNA Synthesis

2013 ◽  
Vol 135 (17) ◽  
pp. 6384-6387 ◽  
Author(s):  
Hailey L. Gahlon ◽  
W. Bernd Schweizer ◽  
Shana J. Sturla
Keyword(s):  
Dna Synthesis ◽  
Base Pair ◽  
Size And Shape

DNA Base Identification by Electron Microscopy

2012 ◽  
Vol 18 (5) ◽  
pp. 1049-1053 ◽  
Author(s):  
David C. Bell ◽  
W. Kelley Thomas ◽  
Katelyn M. Murtagh ◽  
Cheryl A. Dionne ◽  
Adam C. Graham ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Base Pair ◽  
Heavy Atom ◽  
Genomic Research ◽  
Biological Research ◽  
Template Molecule ◽  
Dna Molecules ◽  
Base Pairs ◽  
Dna Molecule ◽  
Dna Base

AbstractAdvances in DNA sequencing, based on fluorescent microscopy, have transformed many areas of biological research. However, only relatively short molecules can be sequenced by these technologies. Dramatic improvements in genomic research will require accurate sequencing of long (>10,000 base-pairs), intact DNA molecules. Our approach directly visualizes the sequence of DNA molecules using electron microscopy. This report represents the first identification of DNA base pairs within intact DNA molecules by electron microscopy. By enzymatically incorporating modified bases, which contain atoms of increased atomic number, direct visualization and identification of individually labeled bases within a synthetic 3,272 base-pair DNA molecule and a 7,249 base-pair viral genome have been accomplished. This proof of principle is made possible by the use of a dUTP nucleotide, substituted with a single mercury atom attached to the nitrogenous base. One of these contrast-enhanced, heavy-atom-labeled bases is paired with each adenosine base in the template molecule and then built into a double-stranded DNA molecule by a template-directed DNA polymerase enzyme. This modification is small enough to allow very long molecules with labels at each A-U position. Image contrast is further enhanced by using annular dark-field scanning transmission electron microscopy (ADF-STEM). Further refinements to identify additional base types and more precisely determine the location of identified bases would allow full sequencing of long, intact DNA molecules, significantly improving the pace of complex genomic discoveries.

scholarly journals Nonreciprocal exchanges of information between DNA duplexes coinjected into mammalian cell nuclei.

1985 ◽  
Vol 5 (1) ◽  
pp. 59-69 ◽  
Author(s):  
K R Folger ◽  
K Thomas ◽  
M R Capecchi
Keyword(s):  
Homologous Recombination ◽  
Dna Sequences ◽  
Plasmid Dna ◽  
Mammalian Cells ◽  
Recombinant Dna ◽  
Dna Duplexes ◽  
Dna Molecules ◽  
Cell Nuclei ◽  
High Concentration ◽  
Length Polymorphisms

We have examined the mechanism of homologous recombination between plasmid molecules coinjected into cultured mammalian cells. Cell lines containing recombinant DNA molecules were obtained by selecting for the reconstruction of a functional Neor gene from two plasmids that bear different amber mutations in the Neor gene. In addition, these plasmids contain restriction-length polymorphisms within and near the Neor gene. These polymorphisms did not confer a selectable phenotype but were used to identify and categorize selected and nonselected recombinant DNA molecules. The striking conclusion from this analysis is that the predominant mechanism for the exchange of information between coinjected plasmid molecules over short distances (i.e., less than 1 kilobase) proceeds via nonreciprocal homologous recombination. The frequency of homologous recombination between coinjected plasmid molecules in cultured mammalian cells is extremely high, approaching unity. We demonstrate that this high frequency requires neither a high input of plasmid molecules per cell nor a localized high concentration of plasmid DNA within the nucleus. Thus, it appears that plasmid molecules, once introduced into the nucleus, have no difficulty seeking each other out and participating in homologous recombination even in the presence of a vast excess of host DNA sequences. Finally, we show that most of the homologous recombination events occur within a 1-h interval after the introduction of plasmid DNA into the cell nucleus.

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