Intermediate stages in enzymatic replication of bacteriophage fd duplex DNA.

Geider, Klaus; Bäumel, Irmtraud; Meyer, Thomas

doi:10.1016/s0021-9258(20)65168-7

Cited by 29 publications

(3 citation statements)

References 33 publications

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“…Nicking of supercoiled duplex RF DNA releases DNA gyrase-imposed torsion, leading to a circular relaxed form. Unsupercoiled phage genome is not a template for phage gene expression [ 60 ]. This may more strongly impact some phage proteins such as pVIII, which are required in high concentrations and continually depleted during progeny virion production.…”

Section: Resultsmentioning

confidence: 99%

Construction and characterization of a novel miniaturized filamentous phagemid for targeted mammalian gene transfer

2023

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Background As simplistic proteinaceous carriers of genetic material, phages offer great potential as targeted vectors for mammalian transgene delivery. The filamentous phage M13 is a single-stranded DNA phage with attractive characteristics for gene delivery, including a theoretically unlimited DNA carrying capacity, amenability to tropism modification via phage display, and a well-characterized genome that is easy to genetically modify. The bacterial backbone in gene transfer plasmids consists of elements only necessary for amplification in prokaryotes, and, as such, are superfluous in the mammalian cell. These problematic elements include antibiotic resistance genes, which can disseminate antibiotic resistance, and CpG motifs, which are inflammatory in animals and can lead to transgene silencing. Results Here, we examined how M13-based phagemids could be improved for transgene delivery by removing the bacterial backbone. A transgene cassette was flanked by isolated initiation and termination elements from the phage origin of replication. Phage proteins provided in trans by a helper would replicate only the cassette, without any bacterial backbone. The rescue efficiency of “miniphagemids” from these split origins was equal to, if not greater than, isogenic “full phagemids” arising from intact origins. The type of cassette encoded by the miniphagemid as well as the choice of host strain constrained the efficiency of phagemid rescue. Conclusions The use of two separated domains of the f1 ori improves upon a single wildtype origin while still resulting in high titres of miniphagemid gene transfer vectors. Highly pure lysates of miniaturized phagemids could be rapidly obtained in a straightforward procedure without additional downstream processing.

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Section: Resultsmentioning

confidence: 99%

Construction and characterization of a novel miniaturized filamentous phagemid for targeted mammalian gene transfer

2023

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“…[ 81 ] The free 3′‐hydroxyl end of the nick serves as a primer for ssDNA synthesis, which proceeds through displacement of the nicked strand as ssDNA by DNA polymerase III and the rep helicase in the presence of single stranded DNA binding protein (SSB). [ 82–83 ] ssDNA replication terminates by pII cleavage of ssDNA at a second downstream nicking site. [ 84 ] The ssDNA is circularized, bound by pV proteins for transport to the cell membrane, and wrapped by the major phage coat and capsid proteins during extrusion of the phage particle.…”

Section: Introductionmentioning

confidence: 99%

“…[1][2][3][4][5][6][7] Their ultrahigh surface areas and ultralow densities make them ideal scaffolds to create single stranded DNA binding protein (SSB). [82][83] ssDNA replication terminates by pII cleavage of ssDNA at a second downstream nicking site. [84] The ssDNA is circularized, bound by pV proteins for transport to the cell membrane, and wrapped by the major phage coat and capsid proteins during extrusion of the phage particle.…”

Section: Introductionmentioning

confidence: 99%

Genetic Control of Aerogel and Nanofoam Properties, Applied to Ni–MnO_x Cathode Design

Cha

Tsedev

Ransil

et al. 2021

Adv Funct Materials

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Aerogels are ultralight porous materials whose matrix structure can be formed by interlinking 880 nm long M13 phage particles. In theory, changing the phage properties would alter the aerogel matrix, but attempting this using the current production system leads to heterogeneous lengths. A phagemid system that yields a narrow length distribution that can be tuned in 0.3 nm increments from 50 to 2500 nm is designed and, independently, the persistence length varies from 14 to 68 nm by mutating the coat protein. A robotic workflow that automates each step from DNA construction to aerogel synthesis is used to build 1200 aerogels. This is applied to compare Ni-MnO x cathodes built using different matrixes, revealing a pareto-optimal relationship between performance metrics. This work demonstrates the application of genetic engineering to create "tuning knobs" to sweep through material parameter space; in this case, toward creating a physically strong and high-capacity battery.

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Sequence Analysis of the Small CrypticXanthomonas campestrispv.vesicatoriaPlasmid pXV64 Encoding a Rep Protein Similar to Gene II Protein of Phage I2-2

Weng

Fan

Tseng

et al. 1997

Biochemical and Biophysical Research Communications

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Intermediate stages in enzymatic replication of bacteriophage fd duplex DNA.

Cited by 29 publications

References 33 publications

Construction and characterization of a novel miniaturized filamentous phagemid for targeted mammalian gene transfer

Construction and characterization of a novel miniaturized filamentous phagemid for targeted mammalian gene transfer

Genetic Control of Aerogel and Nanofoam Properties, Applied to Ni–MnO_x Cathode Design

Sequence Analysis of the Small CrypticXanthomonas campestrispv.vesicatoriaPlasmid pXV64 Encoding a Rep Protein Similar to Gene II Protein of Phage I2-2

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Intermediate stages in enzymatic replication of bacteriophage fd duplex DNA.

Cited by 29 publications

References 33 publications

Construction and characterization of a novel miniaturized filamentous phagemid for targeted mammalian gene transfer

Construction and characterization of a novel miniaturized filamentous phagemid for targeted mammalian gene transfer

Genetic Control of Aerogel and Nanofoam Properties, Applied to Ni–MnOx Cathode Design

Sequence Analysis of the Small CrypticXanthomonas campestrispv.vesicatoriaPlasmid pXV64 Encoding a Rep Protein Similar to Gene II Protein of Phage I2-2

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Genetic Control of Aerogel and Nanofoam Properties, Applied to Ni–MnO_x Cathode Design