Genetic modification, characterization, and co-infection of Franken Sphingomonas and Anaplasma phagocytophilum in tick cells

STAR Protoc. 2023 Sep 15;4(3):102557. doi: 10.1016/j.xpro.2023.102557. Epub 2023 Sep 15.

Abstract

Paratransgenesis through genetic manipulation of symbiotic or commensal microorganisms has been proposed as an effective and environmentally sound approach for the control of vector-borne diseases, including tick bite-related pathologies, and reducing pathogen transmission. Here, we present a protocol for Sphingomonas transformation with Anaplasma phagocytophilum major surface protein 4 and heat shock protein 70. We describe a step-by-step protocol for in vitro study of interactions between transformed Franken Sphingomonas and Ixodes scapularis ISE6 tick cells during A. phagocytophilum infection. For complete details on the use and execution of this protocol, please refer to Mazuecos et al. (2023).1.

Keywords: Biotechnology and Bioengineering; Microbiology.

MeSH terms

  • Anaplasma phagocytophilum* / genetics
  • Animals
  • Coinfection*
  • HSP70 Heat-Shock Proteins / metabolism
  • Ixodes* / genetics
  • Ixodes* / metabolism
  • Sphingomonas* / genetics

Substances

  • HSP70 Heat-Shock Proteins