Bioconjugation of Bacteriophage Pf1 and Extension to Pf1-Based Bionanomaterials

2020 ◽  
Vol 16 ◽  
Author(s):  
Taylor Urquhart ◽  
Bradley Howie ◽  
Lei Zhang ◽  
Kam Tong Leung ◽  
John F. Honek
Keyword(s):  
Coat Protein ◽  
Bulk Material ◽  
Primary Amines ◽  
Wild Type ◽  
Macromolecular Assembly ◽  
Filamentous Bacteriophages ◽  
Functional Biomaterials ◽  
Pf1 Bacteriophage ◽  
Tyrosine Modification ◽  
Single Reaction

Background: Filamentous bacteriophages such as M13 are an important class of macromolecular assembly, rich in chemical moieties that can be used to impart modifiable positions at the nanoscale. Objective: To explore the structurally more complex Pf1 bacteriophage with respect to a diverse set of bioconjugation reactions and to prepare novel fluorescently-labelled Pf1-based composite biomembranes for future applications in areas such as nanoporous filtration biofilms and photoconducting nanocomposite materials. Methods: Pf1 was characterized with respect to amine (N-terminal, Gly1 and Lys20), carboxylate (aspartate, glutamate), and aromatic (tyrosine) modification and its extension to the creation of functional biomaterials. Modification with an amine reactive fluorophore was carried out with Pf1. Results: The reaction profiles between M13 and Pf1 differ, with M13 capable of modification at two primary amines on its major coat protein, while Pf1 is capable of a single reaction per coat protein. Subsequent to the production of dyefunctionalized Pf1, a biocomposite of wild type and functionalized Pf1 could be fabricated into a bulk material by glutaraldehyde (amine-reactive) crosslinking. These biomaterials were characterized by scanning electron and confocal microscopy, showing a distribution of patches of functionalized Pf1 within the main Pf1 construct. Conclusion: The current study provides a framework for future fabrication of advanced bionanomaterials based on the Pf1 bacteriophage.

scholarly journals Comparative reactions of recombinant papaya ringspot viruses with chimeric coat protein (CP) genes and wild-type viruses on CP-transgenic papaya

2001 ◽  
Vol 82 (11) ◽  
pp. 2827-2836 ◽  
Author(s):  
Chu-Hui Chiang ◽  
Ju-Jung Wang ◽  
Fuh-Jyh Jan ◽  
Shyi-Dong Yeh ◽  
Dennis Gonsalves
Keyword(s):  
Coat Protein ◽  
Sequence Similarity ◽  
Transcriptional Gene Silencing ◽  
Papaya Ringspot Virus ◽  
Wild Type ◽  
Coding Region ◽  
Transgenic Papaya ◽  
Cp Gene ◽  
Post Transcriptional Gene Silencing

Transgenic papaya cultivars SunUp and Rainbow express the coat protein (CP) gene of the mild mutant of papaya ringspot virus (PRSV) HA. Both cultivars are resistant to PRSV HA and other Hawaii isolates through homology-dependent resistance via post-transcriptional gene silencing. However, Rainbow, which is hemizygous for the CP gene, is susceptible to PRSV isolates from outside Hawaii, while the CP-homozygous SunUp is resistant to most isolates but susceptible to the YK isolate from Taiwan. To investigate the role of CP sequence similarity in overcoming the resistance of Rainbow, PRSV HA recombinants with various CP segments of the YK isolate were constructed and evaluated on Rainbow, SunUp and non-transgenic papaya. Non-transgenic papaya were severely infected by all recombinants, but Rainbow plants developed a variety of symptoms. On Rainbow, a recombinant with the entire CP gene of YK caused severe symptoms, while recombinants with only partial YK CP sequences produced a range of milder symptoms. Interestingly, a recombinant with a YK segment from the 5′ region of the CP gene caused very mild, transient symptoms, whereas recombinants with YK segments from the middle and 3′ parts of the CP gene caused prominent and lasting symptoms. SunUp was resistant to all but two recombinants, which contained the entire CP gene or the central and 3′-end regions of the CP gene and the 3′ non-coding region of YK, and the resulting symptoms were mild. It is concluded that the position of the heterologous sequences in the recombinants influences their pathogenicity on Rainbow.

scholarly journals Restoration of Wild-Type Virus by Double Recombination of Tombusvirus Mutants with a Host Transgene

1999 ◽  
Vol 12 (2) ◽  
pp. 153-162 ◽  
Author(s):  
Marise Borja ◽  
Teresa Rubio ◽  
Herman B. Scholthof ◽  
Andrew O. Jackson
Keyword(s):  
Coat Protein ◽  
Transgenic Plants ◽  
Coat Protein Gene ◽  
Protein Gene ◽  
Wild Type Virus ◽  
Type Virus ◽  
Virus Infections ◽  
Wild Type ◽  
Transgenic Lines ◽  
Double Recombination

Nicotiana benthamiana plants transformed with the coat protein gene of tomato bushy stunt virus (TBSV) failed to elicit effective virus resistance when inoculated with wild-type virus. Subsequently, R1 and R2 progeny from 13 transgenic lines were inoculated with a TBSV mutant containing a defective coat protein gene. Mild symptoms typical of those elicited in nontransformed plants infected with the TBSV mutant initially appeared. However, within 2 to 4 weeks, up to 20% of the transgenic plants sporadically began to develop the lethal syndrome characteristic of wild-type virus infections. RNA hybridization and immunoblot analyses of these plants and nontransformed N. benthamiana inoculated with virus from the transgenic lines indicated that wild-type virus had been regenerated by a double recombination event between the defective virus and the coat protein transgene. Similar results were obtained with a TBSV deletion mutant containing a nucleotide sequence marker, and with a chimeric cucumber necrosis virus (CNV) containing the defective TBSV coat protein gene. In both cases, purified virions contained wild-type TBSV RNA or CNV chimeric RNA derived by recombination with the transgenic coat protein mRNA. These results thus demonstrate that recombinant tombusviruses can arise frequently from viral genes expressed in transgenic plants.

scholarly journals Ratio of mutated versus wild-type coat protein sequences inPepino mosaic virusdetermines the nature and severity of yellowing symptoms on tomato plants

2013 ◽  
Vol 14 (9) ◽  
pp. 923-933 ◽  
Author(s):  
Beata Hasiów-Jaroszewska ◽  
Anneleen Paeleman ◽  
Nelia Ortega-Parra ◽  
Natasza Borodynko ◽  
Julia Minicka ◽  
...  
Keyword(s):  
Coat Protein ◽  
Protein Sequences ◽  
Wild Type ◽  
Tomato Plants

scholarly journals Recombination with Host Transgenes and Effects on Virus Evolution: An Overview and Opinion

1999 ◽  
Vol 12 (2) ◽  
pp. 87-92 ◽  
Author(s):  
Teresa Rubio ◽  
Marise Borja ◽  
Herman B. Scholthof ◽  
Andrew O. Jackson
Keyword(s):  
Coat Protein ◽  
Protein Gene ◽  
Virus Disease ◽  
Virus Evolution ◽  
Wild Type Virus ◽  
Negative Consequences ◽  
Wild Type ◽  
Sources Of Resistance ◽  
Double Recombination ◽  
Virus Mutant

This commentary relates to the work by M. Borja et al. (M. Borja, T. Rubio, H. B. Scholthof, and A. O. Jackson, MPMI 12:153-162, 1999) that shows that wild-type virus can be restored frequently by double recombination events between a tomato bushy stunt virus mutant with deletions inactivating the coat protein gene and a coat protein transgene. Here, we focus on evidence suggesting that new viruses might evolve via recombination with transgenes used for disease resistance, and discuss the potential effects of widespread use of these sources of resistance on virus evolution. We argue that the benefits arising from using transgenic sources of resistance for virus disease control outweigh potential negative consequences of evolution of novel hybrid viruses with destructive disease potential.

scholarly journals Characterization of a Major Bacillus anthracis Spore Coat Protein and Its Role in Spore Inactivation

2004 ◽  
Vol 186 (8) ◽  
pp. 2413-2417 ◽  
Author(s):  
Ho-San Kim ◽  
D. Sherman ◽  
F. Johnson ◽  
A. I. Aronson
Keyword(s):  
Coat Protein ◽  
Bacillus Anthracis ◽  
Spore Coat ◽  
Wild Type ◽  
Gene Encoding ◽  
Promising Target ◽  
Section Electron ◽  
Group A ◽  
Dark Staining ◽  
Spore Coat Protein

ABSTRACT A major Bacillus anthracis spore coat protein of 13.4 kDa, designated Cotα, was found only in the Bacillus cereus group. A stable ca. 30-kDa dimer of this protein was also present in spore coat extracts. Cotα, which is encoded by a monocistronic gene, was first detected late in sporulation, consistent with a σK-regulated gene. On the basis of immunogold labeling, the protein is in the outer spore coat and absent from the exosporium. In addition, disruption of the gene encoding Cotα resulted in spores lacking a dark-staining outer spore coat in thin-section electron micrographs. The mutant spores were stable upon heating or storage, germinated at the same rate as the wild type, and were resistant to lysozyme. They were, however, more sensitive than the wild type to phenol, chloroform, and hypochlorite but more resistant to diethylpyrocarbonate. In all cases, resistance or sensitivity to these reagents was restored by introducing a clone of the cotα gene into the mutant. Since Cotα is an abundant outer spore coat protein of the B. cereus group with a prominent role in spore resistance and sensitivity, it is a promising target for the inactivation of B. anthracis spores.

Novel Tyrosine Markers in Raman Spectra of Wild-Type and Mutant (Y21M and Y24M) Ff Virions Indicate Unusual Environments for Coat Protein Phenoxyls

Biochemistry ◽  
1994 ◽  
Vol 33 (5) ◽  
pp. 1037-1042 ◽  
Author(s):  
Stacy A. Overman ◽  
Kelly L. Aubrey ◽  
Nelson S. Vispo ◽  
Gianni Cesareni ◽  
George J. Thomas
Keyword(s):  
Coat Protein ◽  
Raman Spectra ◽  
Wild Type
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