An improved genetic system for bioengineering buoyant gas vesicle nanoparticles from Haloarchaea

BMC Biotechnology
Shiladitya DasSarmaBarbara Smith

Abstract

Gas vesicles are hollow, buoyant organelles bounded by a thin and extremely stable protein membrane. They are coded by a cluster of gvp genes in the halophilic archaeon, Halobacterium sp. NRC-1. Using an expression vector containing the entire gvp gene cluster, gas vesicle nanoparticles (GVNPs) have been successfully bioengineered for antigen display by constructing gene fusions between the gvpC gene and coding sequences from bacterial and viral pathogens. To improve and streamline the genetic system for bioengineering of GVNPs, we first constructed a strain of Halobacterium sp. NRC-1 deleted solely for the gvpC gene. The deleted strain contained smaller, more spindle-shaped nanoparticles observable by transmission electron microscopy, confirming a shape-determining role for GvpC in gas vesicle biogenesis. Next, we constructed expression plasmids containing N-terminal coding portions or the complete gvpC gene. After introducing the expression plasmids into the Halobacterium sp. NRC-1 ΔgvpC strain, GvpC protein and variants were localized to the GVNPs by Western blotting analysis and their effects on increasing the size and shape of nanoparticles established by electron microscopy. Finally, a synthetic gene coding for Gaussia pr...Continue Reading

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Citations

Sep 10, 2015·Vaccines·Shiladitya DasSarma, Priya DasSarma
Jun 29, 2016·ACS Nano·Anupama LakshmananMikhail G Shapiro
Jan 20, 2017·Nature Protocols·Glen M DeLoidPhilip Demokritou
Dec 4, 2020·Biotechnology Advances·Kevin PfeiferSimon K-M R Rittmann
Jan 11, 2017·Molecular Pharmaceutics·Abhay U AndarShiladitya DasSarma

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Methods Mentioned

BETA
transmission electron microscopy
PCR
electron microscopy
antigen display
electrophoresis
Assay

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