Design of pVAXhTPA-BB0405 DNA Vaccine

The pVAXhTPA-BB0405 DNA vaccine was designed as described before in Wagemakers et al.^{10 (link)}. In the BB0405 gene sequence of B. burgdorferi B31 (NCBI Reference Sequence: NC_001318.1) the 18aa signal sequence (predicted by SignalP 4.0 web-based software, CBS, Lyngby, Denmark) was replaced with the human tissue plasminogen activator (hTPA) signal sequence (genbank AAA61213.1)^{26 (link)}. The resulting sequence was codon-optimized to mouse tRNA usage with Java Codon Adaptation tool (Braunschweig, Germany)^{27 (link)}. At the 5′ end a BamH1 and a Kozak sequence were added, and at the 3′ end a sequence encoding a double stop codon and a Xho1 were added. The insert was synthesized (BaseClear, Leiden, The Netherlands) and ligated into a BamH1/Xho1 restricted empty pVAX vector (Invitrogen, Carlsbad, CA, USA). The plasmid was amplified using a Nucleobond Xtra EF kit (Macherey-Nagel, Düren, Germany) and resuspended in DNase free water.

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Klouwens M.J., Trentelman J.J., Ersoz J.I., Nieves Marques Porto F., Sima R., Hajdusek O., Thakur M., Pal U, & Hovius J.W. (2021). Investigating BB0405 as a novel Borrelia afzelii vaccination candidate in Lyme borreliosis. Scientific Reports, 11, 4775.

Publication 2021

BamH1/Xho1 restricted empty pVAX vector Nucleobond Xtra EF kit

Adaptation Codon Dna vaccine Dnase Gene Human Mouse Plasmid Signal sequence Stop codon Tissue plasminogen activator Trna Vector

Corresponding Organization : Academic Medical Center

Other organizations : Institute of Parasitology, Czech Academy of Sciences, Biology Centre, Virginia–Maryland College of Veterinary Medicine, University of Maryland, College Park

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Variable analysis

independent variables

Replacement of the 18aa signal sequence (predicted by SignalP 4.0 web-based software, CBS, Lyngby, Denmark) in the BB0405 gene sequence of B. burgdorferi B31 (NCBI Reference Sequence: NC_001318.1) with the human tissue plasminogen activator (hTPA) signal sequence (genbank AAA61213.1)
Codon optimization of the resulting sequence to mouse tRNA usage with Java Codon Adaptation tool (Braunschweig, Germany)

dependent variables

Not explicitly mentioned

control variables

Not explicitly mentioned

controls

Positive control: Not mentioned
Negative control: Not mentioned

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