Journal of Microbiology

Research Support, Non-U.S. Gov'ts

Identification of Porcine Endogenous Retrovirus (PERV) packaging sequence and development of PERV packaging viral vector system: Jiwon Choi , Hoon-mi Kim , Jong Kwang Yoon , Yeondong Cho , Hee-Jung Lee , Kang Chang Kim , Chang-Kyu Kim , Gye-Woong Kim , Young Bong Kim; J. Microbiol. 2015;53(5):348-353. Published online May 3, 2015; DOI: https://doi.org/10.1007/s12275-015-5134-0

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Studies of the retroviruses have focused on the specific interaction of the nucleocapsid protein with a packaging signal in the viral RNA as important for this selectivity, but the packaging signal in porcine endogenous retrovirus (PERV) has not been defined. Herein, we identified and analyzed this packaging signal in PERV and found hairpin structures with conserved tetranucleotides in their loops and nucleocapsid recognition sequences; both of which are key elements in the viral packaging signal of MLV. We evaluated packaging efficiency of sequence variants isolated from viral and proviral integrated genomes. All viral packaging sequences (Ψ) were identical, while five distinct packaging sequences were identified from proviral sources. One proviral sequence (Ψ1) was identical to that of the viral Ψ and had the highest packaging efficiency. Three variants (Ψ2, Ψ3, Ψ4) maintained key elements of the viral packaging signal, but had nucleotide replacements and consequently demonstrated reduced packaging efficiency. Despite of the same overall hairpin structure, the proviral variant (Ψ5) had only one GACG sequence in the hairpin loop and showed the lowest packaging efficiency other than ΔΨ, in which the essential packaging sequence was removed. This result, thus, defined the packaging sequences in PERV and emphasized the importance of nucleotide sequence and RNA structure in the determination of packaging efficiency. In addition, we demonstrate efficient infection and gene expression from the PERVbased viral vector, which may serve as a novel alternative to current retroviral expression systems.

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Porcine Endogenous Retrovirus (PERV) – Molecular Structure and Replication Strategy in the Context of Retroviral Infection Risk of Human Cells
Krzysztof Łopata, Emilia Wojdas, Roman Nowak, Paweł Łopata, Urszula Mazurek
Frontiers in Microbiology.2018;[Epub] CrossRef

Isolation and Characterization of the Smallest Bacteriophage P4 Derivatives Packaged into P4-Size Head in Bacteriophage P2-P4 System: Kyoung-Jin Kim , Jaeho Song; J. Microbiol. 2006;44(5):530-536.; DOI: https://doi.org/2444 [pii]

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Abstract: Bacteriophage P4, a satellite phage of coliphage P2, is a very useful experimental tool for the study of viral capsid assembly and cos-cleavage. For an in vitro cos-cleavage reaction study of the P2-P4 system, new shortened and selectable markers containing P4 derivative plasmids were designed as a substrate molecules. They were constructed by swapping the non-essential segment of P4 DNA for either the kanamycin resistance (kmr) gene or the ampicillin resistance (apr) gene. The size of the genomes of the resulting markers were 82% (P4 ash8 delRI:: kmr) and 79% (P4 ash8 delRI:: apr) of the wild type P4 genome. To determine the lower limit of genome size that could be packaged into the small P4-size head, these shortened P4 plasmids were converted to phage particles with infection of the helper phage P2. The conversion of plasmid P4 derivatives to bacteriophage particles was verified by the heat stability test and the burst size determination experiment. CsCl buoyant equilibrium density gradient experiments confirmed not only the genome size of the viable phage form of shortened P4 derivatives, but also their packaging into the small P4-size head. P4 ash8 delRI:: apr turned out to be the smallest P4 genome that can be packaged into P4-sized head.

Characterization and Identification of the Bacteriophage P4 Mutant Suppressin sir Mutations of Bacteriophage P2: Kim, Kyoung Jin , Sunshine, Melvin G. , Six, Erich W.; J. Microbiol. 1998;36(4):262-265.

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Abstract: Bacteriophage P4 ost1 was isolated as a suppressor mutant of P2 sir3 and identified by restriction enzyme site analysis. The mutant DNA turned out to be an imperfect P4 trimer containing deletions. It was suggested that the deletion resulted from int-mediated site-specific recombination. CsCl equilibrium density gradient experiment confirmed the genome size of P4 ostl.

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